relat act 2004

RELATÓRIO DE ACTIVIDADES
DO CIBO/IPATIMUP
2004
1
ÍNDICE
PÁG.
1.
INTRODUÇÃO
2.
RELATÓRIO DA AVALIAÇÃO DO IPATIMUP PELO PAINEL
NOMEADO PELA FCT (OUTUBRO DE 2004)
3.
2
4
INVESTIGAÇÃO CIENTÍFICA
13
3a. Cancer Biology
14
3b. Cancer Genetics
17
3c. Carcinogenesis
20
3d. Genetics, Evolution and Pathology
22
3e. Molecular Targets in Cancer
24
3f. Population Genetics
30
3g. Tumour Evolution and Development
37
4.
EDUCAÇÃO CONTÍNUA E DIFUSÃO CIENTÍFICA
38
5.
SERVIÇO À COMUNIDADE
41
5a. UPS
42
5b. UPSi
48
5c. UPSs
50
6.
DOUTORAMENTOS
52
7.
TRABALHOS PUBLICADOS OU ACEITES PARA PUBLICAÇÃO
56
7a. Artigos científicos
57
7b. Capítulos de livros
62
2
1. INTRODUÇÃO
3
Dando seguimento às repetidas sugestões do External Advisory Council do IPATIMUP, reforçadas em Outubro
de 2004 pelo Painel encarregado pela FCT de proceder à Avaliação Intercalar do Instituto, inauguramos este
ano um novo formato de Relatório de Actividades.
As diferenças mais pronunciadas dizem respeito à parte científica que passará a ser constituída por relatórios
dos sete Grupos de Investigação escritos em inglês, com identificação das linhas de pesquisa e dos trabalhos
com maior relevância. Os relatórios das Unidades de Prestação de Serviços (em finais de 2004 criámos uma
3ª Unidade, a UPSs, em que o s significa “susceptibilidade”), assim como o relatório da Unidade de Educação
Contínua e Divulgação Científica manterão a mesma estrutura e continuarão a ser escritos em português.
Excepcionalmente, este Relatório inclui o Resultado da Avaliação e as Recomendações do Painel encarregado
pela FCT da Avaliação Intercalar do Instituto. Para além dos aspectos científicos e organizacionais salientados
nesse documento, destacaríamos, como elementos mais marcantes das actividades do IPATIMUP em 2004,
os seguintes factos:
a) Início da construção da nova ala do Instituto
b) Sucesso do Programa de Mecenato Científico ao qual aderiram as seguintes 17 Empresas: Allianz Portugal,
SA; Banco BPI, SA; Bayer Portugal, SA; Bial - Portela & C.ª, SA; GlaxoSmithKline Prod. Farmacêuticos, Lda.;
Merck Sharp & Dohme, Lda.; Millennium BCP; Mota Engil, SGPS, SA; Novartis Farma - Prod. Farmacêuticos,
SA; Portgás - Soc. de Prod. e Distr. de Gás, SA; Portucel SGPS, SA; RAR - Sociedade de Controle (Holding),
SA; Roche Farmacêutica Química, Lda. ; SAG GEST - Soluções Automóvel Globais, SA; Sonae SGPS, SA;
Têxtil Manuel Gonçalves, SA; Unicer SGPS, SA
c) Consolidação da reorganização da actividade científica que passou a estar centrada em 7 Grupos: Cancer
Biology (Coordenador: Paula Soares); Cancer Genetics (Coordenador: Raquel Seruca); Carcinogenesis
(Coordenador: Leonor David); Genetics, Evolution and Pathology (Coordenador: Jorge Rocha); Molecular
Targets in Cancer (Coordenador: Ginesa Garcia-Rostan); Population Genetics (Coordenador: António
Amorim); Tumour Evolution and Development (Coordenador: Luís Costa).
d) Ainda no domínio científico é de salientar a existência de 5 investigadores do IPATIMUP entre os 73
investigadores portugueses de todas as áreas considerados, pela sua produção científica, como excelentes
pelo Ministério da Ciência e Ensino Superior.
e) Crescimento em termos absolutos e relativos da fracção das receitas extra-FCT (sobretudo empresas
privadas e agências internacionais) provenientes de projectos, programas e protocolos de investigação a
realizar pelo IPATIMUP.
f) Expansão das actividades de divulgação científica com a realização de dois Ciclos de 7 Colóquios sobre
Medicina Preventiva do Cancro na Fundação Calouste Gulbenkian(Março/Maio) e na Fundação de Serralves
(Outubro/Dezembro) para mais de 1000 professores do ensino secundário.
g) Reforço da ligação ao IBMC e INEB com a criação de um Program’s Office comum aos três Institutos,
concursos articulados a programas nacionais e internacionais, e uma política sustentada de partilha de
equipamentos, para além da continuação da estreita colaboração na divulgação científica, no Programa
GABBA e em outros tipos de formação pós-graduada.
Nota: O IPATIMUP voltou a contar, em 2004, com o apoio excepcional dos seus Associados Efectivos
(Universidade do Porto, Câmara Municipal do Porto, FLAD, Liga Portuguesa Contra o Cancro, CCRN, IGMJM
e Cruz Vermelha Portuguesa) e Aderentes (Faculdade de Medicina, Faculdade de Ciências, ICBAS,
F.Ciências da Nutrição, F.Medicina Dentária, F.Farmácia, H.S. João e IPO-Porto), assim como da Fundação
Calouste Gulbenkian e da Fundação de Serralves.
4
2.
RELATÓRIO DA AVALIAÇÃO DO IPATIMUP PELO PAINEL NOMEADO PELA
FCT (OUTUBRO DE 2004)
5
Relatório da Avaliação do Laboratorio Associado
Instituto de Patologia e Imunologia Molecular da Universidade do
Porto – IPATIMUP
19 de Outubro de 2004
Painel de avaliação:
Fernando M. S. Tomé, M.D., Ph.D.
João P. Pulido Valente Monjardino, M.D., Ph.D.
Fernando H. Lopes da Silva, M.D., Ph.D.
First Part according to the list of items of the FCT guidelines
Quality and relevance of the scientific activity of IPATIMUP.
•
International recognition of the research activity: namely the number of citations
per publication, international awards; invited oral presentations (plenary lectures,
keynote lectures):
The publication of the last period is of high quality with a strong emphasis on top journals
in the fields of gastroenterology, oncogeny, pathology and human and molecular genetics.
The publications profile has become stronger with an increasing share of publications in
journals with a high impact factor. Several researchers, both senior and graduate students,
have received prestigious prizes from scientific societies and other institutions. Members of
IPATIMUP participate in Editorial Boards of a large number of international journals,
particularly in the field of pathology and give frequently key-note lectures abroad.
•
Breadth and depth of ongoing and planned research: originality of the research
work and its potential impact:
The main focus of the LA has been on research on gastric, thyroid and breast cancer and
lymphoma, population genetics and cancer susceptibility. This covers a broad area of cancer
research within which the original contributions of IPATIMUP are well recognized. The
recently awarded funds for re-equipment ( MALDI-TOF mass spectrometer) for proteomics
research are most welcome in this context, and should be an important stimulus for
reinforcing the study of cellular and molecular mechanisms of carcinogenesis.
•
Current importance of the research themes: involvement in new emerging areas of
research:
From the above it is clear that IPATIMUP is engaged in innovative research in the field of
carcinogenesis which shows great potential.
• Multidisciplinarity and relevance for other research areas and/or technologies:
The group of researchers includes pathologists, clinicians, and basic scientists in biomedical research. The Institute gives a high priority to translational research.
•
Involvement of young researchers and/or post-docs in the research activity:
6
The cumulative number of Ph.Ds that received training at IPATIMUP has steadily
increased from 1 in 1999 to 45 in June 2004; In addition the Institute has at the moment 8
scientists with Post-Doctoral Fellowships.
• Organization of scientific meetings and advanced training seminars
There are scientific seminars on a regular basis with the contribution of speakers from
outside the Institute and often from abroad. The Institute participates actively in the
graduate course GABBA of the University of Porto in collaboration with IBMC/INEB and
other faculties.
Organization and Scientific environment
• Adequacy of Unit organization and leadership:
The LA has a clear organization with a strong leadership both centrally and at the level of
the 7 Research Units that are led by senior researchers. In addition it also has the Scientific
Outreach Programme under the responsibility of a senior professor. The Institute is in the
process of creating a Unit to take responsibility for the Teaching of the Master and Ph.D.
Programmes. The coordinators of the Units form the Steering Committee that meets
monthly. In the Directory Board the Director is assisted by the heads of the Services
department and the managers. General scientific policy is discussed, in principle monthly,
in the Scientific Council where all collaborators holding a Ph.D. participate.
• Culture of creativity and opportunity/encouragement of young researchers initiative
In the Scientific Council junior and senior researchers of all different groups and
specializations meet and discuss new ideas and lines of research. In addition the weekly
Seminars and Journal Clubs provide excellent opportunities for creative interactions during
which the contribution of younger researchers appears to be actively encouraged.
•
Share of common equipment, facilities and resources between different groups
inside the LA:
In view to its dimension and focus of research IPATIMUP functions as a large research unit
with a strong common infrastructure that is used by all different groups.
•
Level of complementary activities and research interactions: number of common
projects and co-supervision of graduate students:
Many interactions between groups exist. Several researchers participate in common projects
and publish frequently together.
•
Organization of seminars and group meetings: level of involvement/participation in
these events:
See above “Culture of creativity…”
• Internal communication (instruments and level of participation)
See above
Internationalisation
• Joint publications with foreign researchers
There are many publications where researchers of IPATIMUP collaborate with foreign
groups.
7
• Participation in scientific and technological projects with foreign researchers:
The Institute participates in joint projects with several foreign research institutions, namely
of the Netherlands, Belgium, Norway, Spain, Germany, United Kingdom, France, USA and
Brazil..
• Involvement in international networks for higher education and research:
The Institute participates in several international networks dedicated to research and data
banks. Some senior researchers are invited (Honorary) Professors in different Medical
Faculties (China, Spain, Brazil, Norway).
• Interaction with foreign researchers and /or research units abroad:
The activities described above indicate clearly that the Institute has a strong interaction with
the international research community. The researchers of IPATIMUP are regularly invited
to give main lectures at international Congresses and Seminars, In addition the Institute
provides diagnostic services not only in Portugal but also in a large number of other
countries.
• Ability to attract foreign post-docs and other researchers (invited or associated):
IPATIMUP receives regularly many researchers for specialized training from several
countries of the European Union. In addition it also receives researchers for training from
other countries, namely from Brazil, Russia, Ukraine, China and Moçambique.
In our view the IPATIMUP should be encouraged to attract Post-Docs from abroad in order
to strengthen its research potential, provided that it receives the necessary extra funds either
from the countries of origin, from the FCT or from other Organizations.
Human resources and Management
• Recruitment policy.
IPATIMUP has the plan to attract a senior researcher and three technicians. This depends
on receiving the necessary funds from FCT. We support this plan and are of the opinion that
in this way the Institute should advertise widely and try to recruit these new collaborators
from anywhere in the world to inject “new blood” and avoid inbreeding.
• Balance between senior and young researchers.
There is a good balance.
• Programs for post-graduate students and advanced training of young researchers.
As indicated above there are adequate programs for advanced training and many
interactions with foreign researchers in this context.
• Adequacy of safety procedures.
As far as we could understand the Direction takes good care of the implementation of
these procedures.
Technology transfer / Diffusion and valorisation of results.
• Spin-off enterprises.
The spin-off enterprise NEST was created. GENETEST provides services in the field of
genetic diagnostics. The Institute collaborates in several projects jointly with a number of
enterprises.
•
Technological prototypes and patents.
8
No information available.
• Other knowledge and technology transfer activities.
See above
• Research contracts with enterprises.
See above
• Services contracted and provided to external entities.
See above
• Diffusion of scientific culture.
IPATIMUP has a strong programme under the responsibility of the Unit for continuing
education and scientific outreach for the diffusion of scientific culture. This is very valuable
and an important contribution to increase the visibility of the Institute.
Strategic planning and long term perspective.
• Clear definition of objectives and “mission statement”.
From the above it is clear that IPATIMUP has a clear mission and has expressed that in
many publications and in other activities.
• Adequacy of objectives in a national and international context.
IPATIMUP occupies a leading position in the field of oncology and is the laboratory of
reference in gastric oncology not only at the national level but also internationally.
Resources of research activity
Adequacy of resources for the research activity:
Facilities.
Adequate
Library.
New space is almost ready.
Equipment.
The LA received an important re-equipment fund.
Technical support
The LA has a good technical staff but needs new senior technicians for three areas:
“cell lines and cell culture”, “tissue and tumour bank” and “biostatistics”.
Administrative support.
Appears adequate
Funding. This is the big problem. The most important signal that we received was a
‘cri-de-coeur’ regarding the inordinate delays in receiving the approved payments from
FCT under the “financiamento programático”, i.e. the lack of stable financial support. We
realized that this is a grave problem. We are of the opinion that this LA merits an extra
financial stimulus to be able to consolidate the admirable task that it is already fulfilling.
Out of the total funding for 2003, and in addition to the FCT funding (33 %), the LA
9
received 25% from services offered to the community, 34% from specific research projects
and 8% from the University and other sources.
10
Relatório da Avaliação do Laboratorio Associado
Instituto de Patologia e Imunologia Molecular da Universidade do
Porto - IPATIMUP
19 de Outubro de 2004
Painel de avaliação:
Fernando M. S. Tomé, M.D., Ph.D.
João P. Pulido Valente Monjardino, M.D., Ph.D.
Fernando H. Lopes da Silva, M.D., Ph.D.
2nd Part:
Comments and recommendations regarding the activities of the “Laboratório Associado”,
research orientation and organization
General aspects
The original evaluation that this LA was Excellent is being confirmed by the outstanding
development of IPATIMUP. In the past three years this LA has developed as an Excellent
research group doing research in a variety of aspects of basic and clinical Oncology, and
with a special emphasis on gastric carcinogenesis. It constitutes a leading scientific
institution in the Health Sciences area of Portugal, with a clear international impact. The
researchers are being very productive, both at the level of scientific publications of high
value, and of providing high quality diagnostic services, developing outreach programs and
inspired programs of diffusion of scientific culture.
The LA has a clear organization with a strong leadership both at the general level and at the
level of the 7 Research Units that are led by senior researchers. In addition it has also the
Scientific Outreach Program under the responsibility of a senior professor. The Institute is
in the process of creating a Unit to take responsibility for the Teaching Program at the
Master and Ph.D. levels. The coordinators of the Units form the Steering Committee that
meets monthly. In the Directory Board the Director is assisted by the heads of the Services
departments and the managers. General scientific policy is discussed, in principle monthly,
in the Scientific Council where all collaborators holding a Ph.D participate.
IPATIMUP is at the point of incorporating well deserved and necessary new space. The LA
has a small independent External Scientific Board, the critical advice of which has had a
very healthy influence in the development of IPATIMUP. The LA wishes to extend the
Board with new members. We welcome this proposal and favor strengthening of the Board
with the contribution of outstanding foreign, rater than Portuguese scientists.
The recruitment of talented Post-Docs is a high priority. The strengthening of the technical
staff to supervise a Cell Culture facility, to run a Tissue and Tumour Bank and to be in
charge of Biostatistics is an urgent necessity. We suggest that the latter could be realized as
a joint appointment with IBMC/IBED.
Quality and relevance of the scientific activity of IPATIMUP
The latest record of scientific publications shows that they are of high quality with a strong
emphasis on top journals in the fields of gastroenterology, oncogeny, pathology and human
and molecular genetics. The number of publications in SCI journals has been increasing
since the time the LA was recognized. The publications profile has become also stronger
with an increasing share of publications in journals with a high impact factor. Nevertheless
11
there is also a relatively large number of publications in non-SCI journals (mostly in
Portuguese) that correspond mainly to studies carried out in collaboration with clinical
groups. The latter are important for the institution mainly for strategic reasons, but an active
effort to increase the international visibility of these studies should be pursued. Several
(n=7) publications that appeared in the period immediately preceding the constitution of the
LA (1996 – 1999) have reached more than 100 citations in 2004; during the more recent
years the number of SCI publications in journals with an impact factor larger than 6.5,
which can be considered first class in this research field, rose from 7% (2000) to 17%
(2003). Several researchers, at least 11, have papers that obtained a score of citations/year
higher than 10 (according to the ISI Web of Science), what should be considered
outstanding in the field1.
Several researchers, both senior and graduate students, have received prizes from scientific
societies and other institutions. Two awards merit being mentioned specially: the Benjamin
Castleman prize, that was awarded by the United States and Canadian Academy of
Pathology to “the most outstanding paper in the field of human pathology published in
2001” with the contribution of two IPATIMUP researchers Fátima Carneiro and Raquel
Seruca in collaboration with a group of foreign colleagues. The Director M. Sobrinho
Simões received the prestigious Prémio Pessoa in 2004.
Members of IPATIMUP participate in Editorial Boards of a large number of international
journals, particularly in the field of pathology, and give frequently key-note lectures abroad.
Scope of the research themes
The main focus of the LA has been on research on gastric, thyroid and breast cancer and
lymphoma, population genetics and cancer susceptibility. This covers a broad area of cancer
research within which the original contributions of IPATIMUP are well recognized. An
actual question that is being tackled by the scientists at the Institute is whether to widen
their field of expertise to cancers of other organs or to develop the current investigation
further by using cell biology-based complementary experimental approaches. A
combination of both is likely the best strategy, but the latter aspect appears to us an
important priority. To realize this the LA needs to create conditions to enable cell biologyoriented research. The recently awarded funds for re-equipment ( MALDI-TOF mass
spectrometer) for proteomics research are most welcome in this context, and should be an
important stimulus for reinforcing the study of cellular and molecular mechanisms of
carcinogenesis.
The group of researchers is constituted by pathologists, clinicians, and basic scientists in
bio-medical research. The Institute gives a high priority to translational research. This is an
important asset, and it assures the relevance of the know-how developed at IPATIMUP to
other clinical research areas. We should note, however, that a strong base in fundamental
bio-medical research is always necessary to establish significant links with the field of
clinical applications.
Involvement of young researchers and/or post-docs in the research activity:
1
Here we distinguish 4 levels with respect the citations/year scores per paper. The argument for this
classification is the following: a score above 10 corresponds to the level of the Impact factors of the most
influential multidisciplinary Journals and the most important ones in some fields of bio-medical research; a
score between 6.5 and 10 corresponds to the group of excellent specialized Journals in different fields of this
kind of research; a score between 3 and 6.5 corresponds to the Journals that are considered very good
specialized Journals in many bio-medical areas. A score smaller than 3 corresponds to Journals with low
impact. This is, of course, just a way of ordering the impact of various publications, and it depends very much
on research area. A more professional assessment of research quality should take more factors in consideration.
Such a professional quantitative assessment should be made by the FCT in future evaluations.
12
The cumulative number of Ph.Ds. that received training at IPATIMUP has steadily
increased from 1 in 1999 to 45 in June 2004; In addition the Institute has at the moment 8
scientists with Post-Doc Fellowships. This is a valuable achievement. IPATIMUP has the
plan to attract a senior researcher and three technicians. This depends on receiving the
necessary funds from FCT. We support this plan. Furthermore it is important that
IPATIMUP develops a policy to recruit young scientists at the Post-Doc level with
experience in other laboratories, either in foreign countries or in Portugal, that should be
given the opportunity of starting new research units. This would reinforce the dynamism of
the research and help avoiding inbreeding.
In the Scientific Council junior and senior researchers of all different groups and
specializations meet and discuss new ideas and lines of research. In addition the weekly
Seminars and Journal Clubs provide excellent opportunities for creative interactions where
the contribution of younger researchers appears to be actively encouraged. There are
scientific seminars on a regular basis with the contribution of speakers from outside the
Institute and often from abroad. The Institute participates actively in the graduate course
GABBA of the University of Porto in collaboration with IBMC/INEB and other faculties.
IPATIMUP receives regularly many researchers for specialized training from several
countries of the European Union. In addition it receives also researchers for training from
other countries, namely from Brazil, Russia, Ukraine, China and Moçambique.
IPATIMUP has a strong program under the responsibility of the Unit for continuing
education and scientific outreach for the diffusion of scientific culture. This is very valuable
and forms an important element that increases the visibility of the Institute.
Summing up IPATIMUP is an Excellent institution that occupies a leading position in the
field of oncology and is the laboratory of reference in gastric oncology not only at the
national level but also internationally. Funding is the main concern. The most important
signal that we received was a ‘cri-de-coeur’ regarding the solutions of continuity with
respect to receiving the “financiamento programático”, i.e. the lack of stable financial
support. We realized that this is a grave problem. We are of the opinion that this LA merits
an extra financial stimulus to be able to consolidate the admirable task that it is already
fulfilling.
13
3. INVESTIGAÇÃO CIENTÍFICA
3a. Cancer Biology
3b. Cancer Genetics
3c. Carcinogenesis
3d. Genetics, Evolution and Pathology
3e. Molecular Targets in Cancer
3f. Population Genetics
3g. Tumour Evolution and Development
14
Cancer Biology
Coordinator: Paula Soares, BSc, MSc, PhD,
Principal investigators: Clara Sambade, MD, PhD, M. Helena Vasconcelos Meehan, BSc, MSc,PhD, José
Manuel Lopes, MD, PhD, Manuel Sobrinho Simões, MD, PhD,
Post-Doc students: Valdemar Máximo, BSc, PhD; Ana Preto, BSc, PhD; Ana Sofia Rocha, BSc, PhD,
PhD students: Jorge Lima, Patrícia Castro, Vítor Gradíssimo.
MSc students: Raquel Lima, Susana Ribeiro.
Graduate students: - Tália Feijão, Patrícia Pontes, Ana Paula Rebocho, Lígia Gomes, Ricardo Marques,
Ricardo Soares.
Objectives/Goals of the research activity
The broad research interest of the Cancer Biology Group is the molecular etiopathogenesis of human cancer,
with putative application in diagnostics/therapeutics, with an emphasis on thyroid cancer. In addition to this
translational research component, the group has a more basic scientific interest in cell cycle, apoptosis and
motility/invasiveness processes. It is also engaged in some aspects of cancer research which may lead to
validation and/or identification of new molecular targets for cancer treatment (use of cell signalling inhibitors and
downregulation of gene expression with siRNAs ). Besides thyroid tumours the group make also use of
neuroendocrine, breast and haematological malignancies as biological models.
Research Areas
Oncobiology of differentiated thyroid tumours
Radiation induced thyroid tumourigenesis
Oncobiology of familial forms of thyroid and neuroendocrine tumours.
Oncobiology of haematological malignancies.
Validation of molecular targets (namely targets in cancer chemotherapy resistance)
Background and major achievements during 2004
A significant part of our work has been focused on the neoplastic development of differentiated thyroid
carcinomas (ethiopatogenesis and progression), because they represent a particularly original oncobiology
model.
In 2003 we have identified BRAFV600E mutation as a major oncogenic event in PTC (in about 50% of the
cases). The BRAFV600E mutation was specifically associated with the conventional type of PTC as well as
with some variants of PTC displaying a prominent papillary growth pattern. In 2004 we showed that BRAF
mutation is associated with PTC that can progress to undifferentiated carcinoma, since it was found in 30% of
the UC cases (Soares et al, Virchows Arch, 2004 ). We also showed that it was less frequently found in
multicentric tumours (Trovisco et al, Virchows Arch, in press ) thus questioning the association between BRAF
mutation and poor prognosis. Finally we demonstrated it was rarely involved in the pediatric and postChernobyl tumours (Lima et al, JCEM, 2004).
We have also found that a different type of BRAF mutation (K601E) is detected in some of the cases of
Follicular Variant of PTC (7-9%) thus reinforcing the concept of the singularity of FVPTC. We have consolidated
this study by evaluating the presence of PAX8/PPAR rearrangementand ras mutations in FVPTC. The data
obtained shows that FVPTC may be, in molecular terms, more closely related with FTC than with classical PTC
(manuscript in preparation).
In order to further understand the etiopathogenic role of BRAF mutations, we have established an in vitro
system where we could evaluate the biological role (in terms of cellular proliferation, death and invasion) and
the signalling pathways activated by the different BRAF mutations in thyroid cell lines. To achieve this goal we
are using pharmacological kinase inhibitors and BRAF reporter assays.
Aneuploidy is, in the majority of tumour models, a feature associated with aggressive tumours. The two major
histotypes of differentiated thyroid carcinoma display different patterns of DNA content: follicular carcinomas
are clearly aneuploid whereas almost every papillary carcinoma is diploid or quasi –diploid. Applying FISH
analysis to a subset of thyroid benign tumours and carcinomas (fetal type) that frequently show a triploid pattern
we verified that follicular tumorigenesis may follow at least two pathways: one characterized by prominent
15
aneuploidy and numerous chromosomal gains, in which the tumors display a fetal adenoma-like growth pattern,
and another accompanied by less obvious aneuploidy or even quasi-diploidy and dominant chromosome
losses, in which tumors display a common follicular architecture (Castro et al, J Pathol in press).
It is not well understood the role of the mtDNA mutations/variants nor the role of genetic alterations in nuclear
genes that codify for mitochondrial proteins on the etiopathogenesis of mitochondrion-rich (Oncocytic, Hürthle
cell) tumours of the thyroid. We have previously reported that alterations in mitochondrial genes (e.g. ATPase6) can predispose to the development of Hurthe cell tumours. Recently, a gene predisposing to thyroid tumors
with cell oxyphilia (TCO) was mapped to chromosome 19p13.2 in one family with Hürthle cell carcinomas.
GRIM-19 (Gene associated with Retinoid-Interferon-Induced Mortality) a gene located in the 19p13.2 region
has been found to fulfill two roles within the cell: i) It is a member of the interferon- and, a retinoic acid-induced
pathway of cell death, and ii) It is part of the mitochondrial Complex I assembly. It could therefore be involved
both in cell and mitochondrial growth, two of the characteristics features of Hürthle cell tumours. We found
somatic and germline mutations in the above mentioned gene (GRIM-19), in mitochondrion-rich tumours of the
thyroid (Máximo et al, BJCancer, in press). Other nuclear genes potentially relevant to the development of
thyroid tumours have been mapped to chromosome 19 and 17 in familial, as well as in sporadic forms of
Hürthle cell tumours. We have, therefore expanded this study to other mithocondrial related genes localized in
these chromosome regions (PRSS15, NDUFA7, TIMM44, TIMM22 and HCCS1), in order to identify new genes
putatively involved in the etio-pathogenesis of Hürthle cell tumours.
The SDH mitochondrial proteins were implicated in familial forms of neuroendocrine tumours (paraganglioma,
pheocromocitoma), but their role in C-cell derived tumours was unknown. We identified in 2003, in a familial
case of C-cell hyperplasia without ret mutation, a new SDHD gene variation that segregates with the disease.
We are now estimating the frequency of alterations in SDH genes in a series of medullary carcinomas without
ret mutation.
We are also involved in the study of cell adhesion molecules implicated in loss of differentiation and in the
acquisition of invasive proprieties by tumour cells (e.g. cadherins and catenins). We have analyzed members of
a family of thyroid cell lines (KAT) because they express a higher molecular weight (135 kDa) form of Ecadherin at their surface. We found that this aberrant E-cadherin is due to a point mutation in the exon 9 donor
splice site causing skipping of exon 9 with consequent deletion of the corresponding aminoacids of E-cadherin
protein. As a spin-off result, we reported that the various members of the KAT family share this mutation as well
as the same genetic background. We also showed that despite having a disturbed cell-cell adhesion the cells
with the mutated E-cadherin are not invasive unless they suffer an exogenous stimulus (Rocha et al, Thyroid,
2004).
Aiming to address the validation and/or identification of new molecular targets for cancer treatment two
strategies are being followed: the use of cell signalling inhibitors and down regulation of gene expression with
siRNAs. These studies have financial support from NOVARTIS
The resistance of cancer cells to chemotherapeutic drugs may be due to overexpression of antiapoptotic genes
or of the multidrug resistance gene-1 (mdr-1) We verified that downregulation of bcl-2 and xIAP with siRNAs
increased cellular apoptosis in human breast cancer cells (MCF-7 cell line) and enhanced the effects of
chemotherapeutic agents (Lima R et al, Cancer Gene Therapy, 2004). Similar results were obtained when XIAP
was targeted in a chronic myeloid leukaemia cell line (K562) (submitted for publication). Modulation, by siRNA,
of genes from the NF-KB anti-apoptotic dependent pathway have also been address and can lead, as well, to
the identification of new therapeutic targets.
Work plan for 2005
In order to progress in the understanding of the biological role of BRAF activation we will study the effects of
kinase signalling inhibitors (Bay, Gleevec and PD) in thyroid cell lines with and without BRAF mutation, using
for comparative purposes the rates of proliferation and apoptosis.
Even though the prevalence of BRAF activation in thyroid cancer is well known, the prognostic significance of
this alteration remains to be fully elucidated. In collaboration with clinicians from HSJ that have retrospectively
characterized a cohort of patients with differentiated thyroid cancer, we plan to advance in the
clinicopathological characterization of the neoplastic disease in relation to the presence or absence of BRAF
mutation.
The fact that cancers with BRAFV600E mutations do not usually harbour mutations in RET/PTC or RAS
oncogenes, raises the possibility that the coexistence of such activating mutations lead to a proliferative and/or
16
survival disadvantage. Taking this into account, we want to study the biological effect of the coexistence of two
oncogenic activation events BRAF and RET/PTC in thyroid carcinoma cells models. We intend to transfect
BRAFV600E in a cell line with RET/PTC as well as introduce RET/PTC in BRAFV600E positive cell lines.
It is well established the familial occurrence of endocrine and neuroendocrine forms of cancer. With the
collaboration of clinicians from HSJ and IPO, we have started a couple of years ago to collect cases/blood from
suspected familial forms of thyroid cancer (medullary and non-medullary) and other neuroendocrine tumors
(pheochromocytomas and paragangliomas). These families will be evaluated by LOH and linkage studies in
critical genetic regions already pointed out has being involved in familial forms.
The possibility of studying a series of post-Chernobyl tumours lead us to the development of a project that aims
to understand the effect of irradiation in mitochondrial DNA. We verified that irradiated tumours possess
numerous alterations in mtDNA that are already present in the adjacent thyroid parenchyma but not in the
peripheral blood of the patients. These findings raise interesting questions on the effect of the I131 irradiation
on the thyroid as a whole. We plan to perform cloning analysis and large scale sequencing of mtDNA in paired
samples (blood, adjacent thyroid and tumour), in order to verify if the mutated sequences are already present in
all the samples, and, in the affirmative case, if they are present in different proportions.
In 2005 we will initiate a project (supported by UNICER) centred in the identification of natural compounds that
can diminish oxidative stress, evaluated by quantification of alterations in mtDNA, in normal and neoplastic
cells. This project may shed new lights into the identification of putative molecular targets involved in the
oxidative stress.
We intend to continue the validation of new molecular targets for cancer treatment, focusing on anti-apoptotic
proteins and on signal transduction molecules, using both the RNAi and the kinase inhibitors strategies.
Survivin has an important role in cell division (and apoptosis) and can be involved in the mechanism of
aneuploidy in cancer (as well as the other members of the so-called “chromosomal passengers” complex). In a
model of hematopathological malignancy (Hodgkin disease) the proteins of this complex are being analyzed in
order to verify if they are involved in the characteristic multinucleated phenotype present in the disease.
17
Cancer Genetics
Coordinator: Raquel Seruca, MD, PhD
Principal investigators:
Fátima Carneiro, MD, PhD; Fernando Schmitt, MD, PhD; José Carlos Machado, PhD; Céu Figueiredo, PhD;
Gianpaolo Suriano, PhD; Carla Oliveira, PhD.
Post-Doc students: Maria José Oliveira, PhD; Joana Paredes, PhD.
PhD students: Paulo Canedo, Paulo Ferreira, Mónica Botelho, Catarina C Alves, Rita Mateus, Ana Maria
Ferreira, Gonçalo Regalo, António Carlos Ferreira, Fernanda Milanezi, MD.
MSc students: André Albergaria, Silvia Carvalho, Ana Sofia Ribeiro
Graduate students: Sérgia Velho, Cátia Moutinho, Ana Costa, Fábio Pereira.
The research of our group focuses on molecular genetics of 3 common types of epithelial cancer (gastric,
breast, and colorectal carcinoma). We aim at:
1)Identifying individuals at risk of
Gastric cancer: We concentrated our efforts in CDH1 mutation screening in hereditary diffuse gastric cancer
(HDGC) and in genes that were likely to be good alternative candidates. We found one CDH1 and one P53
positive families. We concentrated in IPATIMUP the functional studies of all germline CDH1 missense
mutations detected by the International Gastric Cancer Linkage Consortium (IGCLC). We described for the first
time a new syndrome related to HDGC, where diffuse gastric cancer occurs with concomitant midline
congenital malformations, both diseases harbouring a CDH1 splice-site germline mutation. We extended our
study of identifying a specific genetic profile (of inflammation-related genetic polymorphisms) associated with
risk of development of Helicobacter pylori-associated gastric carcinoma, by analysing in a case-control study
IL6, IL8, IL10, COX2, MIF, TNFA, CTL4 and IFNGR1. Breast cancer: We extended the screening of BRCA1
and BRCA2 in families with aggregation of breast cancer and analyzed negative families for other DNA repair
genes (Rad51, XRCC1, XRCC3, XPD).
2)Identifying pathological features and molecular markers occurring in the setting of familial cancer
We identified a clinicopathologic profile of early-onset gastric carcinoma. The IGCLC decided to concentrate in
IPATIMUP the pathologic analysis of a series of 9 prophylactic gastrectomies from CDH1 mutation carriers
belonging to HDGC. After detailed (topographic and histological) examination of the stomachs, we identified
particular histological features occurring in HDGC and proposed a pathological model of HDGC. Further, the
distribution and size of the cancers in the gastrectomy specimens have clearly indicated that standard
endoscopic screening is unlikely to provide sufficiently sensitive clinical screening for at-risk individuals.
In familial breast cancer we identified a specific basal phenotype (ER and HER-2 negativity and overexpression
of P-cadherin, CK5, and p63) that can be used as a valuable tool for identifying familiar cases within breast
carcinomas.
In HNPCC we verified that BRAFV600E mutation is never found in contrast to its presence in 40% of MSI
sporadic carcinomas allowing introducing this molecular marker in the genetic testing for HNPCC.
3)Identifying signaling pathways mediated by genetic and environmental factors in tumor development
and specific-target therapeutic drugs
Signaling pathways mediated by E-cadherin - We verified an association between the specific location of each
E-cadherin germline missense mutations, cell behavior (morphology and motility) and activation of specific
signaling pathways, namely Rho-GTPases and NF-kB. We established by viral infection human cell lines stably
expressing either wild-type E-cadherin or mutant forms in order to use cDNA microarray and kinase and
phosphatase screenings as broad approaches to identify the common E-cadherin-dependent targets critical for
cell invasion. Signaling pathways mediated by H. pylori - We verified, after co-culture of different H. pylori
strains with gastric cell lines, that strains that are more virulent stimulate cell invasion and motility, and that H.
pylori-induced cell invasion is independent of PI3K activation and is mediated by c-Met tyrosine
phosphorylation. Signaling pathways mediated by IL-6R and its effector C/EBP-beta transcription factor – We
verified that C/EBP-beta overexpression occurs in various stages of gastric cancer development (intestinal
metaplasia, dysplasia and gastric carcinoma) and there is an overlap of expression of C/EBP-beta and COX-2.
We showed, by in vitro assays, that activating and inactivacting C/EBP-beta isoforms (LAP and LIP) lead to a
tight regulation of COX2 expression. Signaling pathways mediated by P-cadherin- We found P-cadherin
expression is regulated by the methylation status of its promoter or ER treatment. We confirmed that the ER
inhibition leads to P-cadherin overexpression. Additionally, we verified that breast cancer cells stably
expressing P-cadherin (by retroviral transduction) are invasive in collagen and Matrigel matrixes and we
18
identified that this functional effect requires the juxtamembrane domain of its cytoplasmic tail. Known targetspecific therapeutic drugs in breast cancer. We established the most reliable screening method to identify
patients with breast carcinomas with HER2/neu amplification that should be treated with Herceptin. We
excluded the potential of Imatinib to treat breast cancer patients due to the lack of c-KIT and PDGFR activating
mutations in breast carcinoma. We verified that EGFR overexpression occurs in metaplastic breast carcinomas
supporting the potential use of EGFR inhibitors (IRESSA) in the treatment of these particular aggressive
carcinomas.
Specific aims of the group for 2005
•
Genetic testing of individuals at risk for gastric carcinoma.
1) Identifying, in collaboration with a specialised centre in S. Paulo- Brasil, new E-cadherin positive HDGCfamilies showing aggregation of cleft-lip;
2) Determining if germline CDH1 large intragenic deletions occur in HDGC families;
3) Determining the existence of a genotype-phenotype correlation associated to germline truncated E-cadherin
protein escaping Nonsense Mediated Decay (NMD).
4) Establishing a cooperative research project with a genetic testing SME, to help us translating the outcome of
the identification of genetic susceptibility factors studies into clinically relevant interventions that will help
decrease the incidence of gastric cancer.
Pathological features and molecular markers occurring in the setting of familial cancer
1) Identifying the second hit mechanisms in tumours from CDH1 positive family members and its relationship
with the disease presentation.
2) Validating the presence of a basal phenotype in a series of breast carcinoma harboring germline mutations
of BRCA genes in order to support these pathological/molecular markers as reliable, fast, and low cost strategy
for the identification of individuals with germline mutations in BRCA genes.
3) Identifying chromosomal alterations crucial for myoepithelial/basal differentiation of the breast by CGH.
Signaling pathways mediated by genetic and environmental factors in tumor development in order to
find new molecular targets for therapeutic intervention.
1) Defining the different expression and protein activation profiles of the human cell lines stably expressing Ecadherin missense mutations to generate new information on the role of E-cadherin deregulation in cell
invasion. Moreover, this study will lead to the identification of alternative genetic events underlying HDGCfamily negative for E-cadherin germline mutations.
2) Determining the cellular effects (proliferation, apoptosis, invasion, motility and wound repair capacity)
mediated by H. pylori virulence factors. These studies will contribute to elucidate the role of H. pylori virulence
factors relevant to carcinogenesis and will lead to the identification the molecular mechanisms underlying those
responses.
3) Clarifying the molecular mechanisms underlying the relationship between polymorphisms associated with
gastric cancer risk and the level of expression of their respective genes.
4) Identifying changes in the expression status of the IL-1R and IL-6/GP130 receptor signal transduction
pathways along the process of gastric carcinogenesis.
6) Determining if inhibition of P-cadherin expression (by RNAi or inhibitory functional antibodies) in ER-negative
breast cancer cells induce ER expression, thus allowing to treat patients, namely BRCA carriers, with hormone
therapy that has being used in patients with ER-positive tumour. 7) Determining the cellular effects (proliferation
and apoptosis) mediated by inhibition of KRAS and BRAF or its downstream targets in the MAPkinase pathway,
using distinct MSI colorectal cell lines with KRAS or BRAF activation treated with pharmacological agents.
Selected references of the group (2003-2005):
1) Carneiro F, Huntsman DG, Smyrk TC, Owen DA, Seruca R, Pharoah P, Caldas C, Sobrinho-Simoes M. Model
of the early development of diffuse gastric cancer in E-cadherin mutation carriers and its implications for patient
screening. J Pathol 203: 681-687, 2004.
We proposed a model of the early development of diffuse gastric cancer in E-cadherin mutation carriers
and described the limitations of standard endoscopic screening in the clinical screening for at-risk
individuals.
2) Machado JC, Figueiredo C, Canedo P, Pharoah P, Carvalho R, Nabais S, Alves CC, Campos ML, van Doorn
LJ, Caldas C, Seruca R, Carneiro F, Sobrinho-Simões M. A pro-inflammatory genetic profile increases the risk
of chronic atrophic gastritis and gastric carcinoma. Gastroenterology 125: 364-371, 2003.
19
We identified a specific genetic profile (of inflammation-related genetic polymorphisms) associated
with risk of development of Helicobacter pylori-associated gastric carcinoma.
3) Oliveira C, Suriano G, Ferreira P, Canedo P, Kaurah P, Mateus AR, Ferreira A, Ferreira AC, Oliveira MJ,
Figueiredo C, Carneiro F, Keller G, Huntsman D, Machado JC, Seruca R. Genetic Screening for Familial
Gastric Cancer. Hereditary Cancer Clinical Practice 2: 51-64, 2004. (Review)
We reviewed all the genetic mutations and risk polymorphism´s associated to familial gastric cancer,
including hereditary diffuse gastric carcinoma.
4) Oliveira C, Westra JL, Arango D, Ollikainen M, Domingo E, Ferreira A, Velho S, Niessen R, Lagerstedt K,
Alhopuro P, Laiho P, Veiga I, Teixeira MR, Ligtenberg M, Kleibeuker JH, Sijmons RH, Plukker JT, Imai K, Lage
P, Hamelin R, Albuquerque C, Schwartz S Jr, Lindblom A, Peltomaki P, Yamamoto H, Aaltonen LA, Seruca R,
Hofstra RM. Distinct patterns of KRAS mutations in colorectal carcinomas according to germline mismatch
repair defects and hMLH1 methylation status. Hum Mol Genet 13: 2303-2311, 2004.
We defined the frequency and type of KRAS and BRAF mutations in HNPCC and in MSI sporadic
colorectal carcinoma
5) Paredes J, Stove C, Stove V, Milanezi F, Van Marck V, Derycke L, Mareel M, Bracke M, Schmitt F. Pcadherin is upregulated by ICI 182,780 in breast cancer cells and promotes invasion via its juxtamembrane
domain. Cancer Res; 64:8309-8317, 2004.
We verified that ER signaling regulates P-cadherin expression and P-cadherin induces breast cancer
cell invasion, being this effect mediated by its juxtamembrane domain
6) Suriano G, Oliveira C, Ferreira P, Machado JC, Bordin MC, De Wever O, Bruyneel EA, Moguilevsky N,
Grehan N, Porter TR, Richards FM, Hruban RH, Roviello F, Huntsman D, Mareel M, Carneiro F, Caldas C,
Seruca R. Identification of CDH1 germline missense mutations associated with functional inactivation of the Ecadherin protein in young gastric cancer probands. Hum Mol Genet 12: 575-582, 2003. .
We described for the first time germline mutations in early-onset diffuse carcinoma without family
history and established an in vitro model to functionally assess the pathogenic role of CDH1 missense
mutations.
7) Suriano G, Oliveira MJ, Huntsman D, Mateus AR, Ferreira P, Casares F, Oliveira C, Carneiro F, Machado
JC, Mareel M, Seruca R. E-cadherin germline missense mutations and cell phenotype: evidences for the
independence of cell invasion on the motile capabilities of the cells. Hum Mol Gen 12: 3007-3016, 2003.
We showed that E-cadherin mediated invasion is not dependent of an increase in cell motility and
distinct signaling pathways mediate the two cellular effects.
8) Frebourg T, Oliveira C, Hochain P, Karam R, Manouvrier S, Graziadio C, Vekemans M, Hartmann A, BaertDesurmont S, Alexandre C, Lejeune S, Dumoulin, Marroni C, Martin C, Castedo S, Lovett M, Winston J,
Machado JC, Attié T, Jabs EW, Cai JL, Triboulet JP, Scotte M, Le Pessot F, Hedouin A, Carneiro F, Blayau M,
Seruca R. Cleft lip/palate and CDH1/E-cadherin mutations in families with hereditary diffuse gastric cancer. J
Med Genet (in press)
We defined a new syndrome with clustering of diffuse gastric cancer and midline malformations with a
CHD1 germline mutation.
20
Carcinogenesis
Coordinator: Leonor David, MD, PhD
Principal investigators:
Fátima Gartner, PhD; Celso Reis, PhD; Raquel Almeida, PhD; Filipe Silva, PhD.
PhD students : Patrícia Mesquita, Jacinta Serpa, Nuno Marcos, Carla Carrilho, MD.
MSc students : Paula Paulo.
Graduate students : José António Almeida, Sandra Pinho.
Objectives/Goals of the research activity - The main objective of the group is to identify alterations of mucins
and mucin glycosylation, associated with gastric carcinoma and precancerous lesions, that may be relevant for
the development of diagnostic and therapeutic strategies. We are also engaged in understanding the molecular
mechanisms involved in the development of such alterations, including the identification of genetic
polymorphisms relevant to identify populations with increased susceptibility to environmental aggressions.
Major achievements during 2004 – Following previous studies of the group on the influence of mucin and
mucin-glycosylation on the susceptibility for Helicobacter pylori infection and gastric diseases thereof, the group
identified two new inactivating mutations of the fucosyltransferase FUTII gene and showed that they are
inactivating in vitro and responsible for a proportion of non-secretors (Paper 1). The group identified
sialyltransferase gene ST6GalNAc-I as the major synthase for the cancer associated antigen Sialyl-Tn,
previously demonstrated by the group as a neo-antigen in pre-neoplastic lesions of the gastric mucosa and as a
structure relevant for invasive and metastatic capacity of carcinomas (Paper 2). The group demonstrated that
the T (Thomsen-Friedenreich) antigen expression is associated, in vivo and in vitro, with large alleles of MUC1
mucin (large VNTR domain) (Paper 3). We have, for the first time, demonstrated a link between protein size
variation and glycosylation, which may be very relevant to clarify previous epidemiological evidence from the
group showing associations of MUC1 VNTR with risk for development of intestinal metaplasia and gastric
carcinoma. We made progress in the characterization of transcriptional regulation of genes associated with the
development of intestinal metaplasia, namely of the intestinal homebox gene CDX2 – we demonstrated an
auto-regulatory CDX2/CDX2 loop in gastric cells and identified OCT-1 as a candidate transcriptional regulator
(unpublished observations). This extends previous work from the group on transcriptional regulation of intestinal
transdifferentiation, using mucins as target genes.
The group was involved during 2004 in a large study of follow-up of a cohort of healthy volunteers from
Estaleiros Navais de Viana do Castelo, which represented a lot of effort for the group: collection of
blood-samples for genetic studies, saliva for secretor status, endoscopies and biopsies, food
questionnaires (all set up in the shipyard). This will allow to settle the relevance of gene-environment
interaction parameters identified in previous studies and also to explore new candidate targets.
The group was also involved in setting up, together with a biotechnology company, a kit for the
diagnosis of intestinal metaplasia based on the identification of alterations of the mucin expression
profile using monoclonal antibodies previously produced by the group.
Selected publications:
1. Serpa J, Mendes N, Reis CA, Santos-Silva F, Almeida R, Le Pendu J, David L: Two new FUT2
(fucosyltransferase 2 gene) missense polymorphisms, 739G-->A and 839T-->C, are partly responsible for nonsecretor status in a Caucasian population from Northern Portugal.Biochem J 383:469-474, 2004
2. Marcos NT, Pinho S, Grandela C, Cruz A, Samyn-Petit B, Harduin-Lepers A, Almeida R, Silva F, Morais V,
Costa J, Kihlberg J, Clausen H, Reis CA: Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis
of the cancer-associated sialyl-Tn antigen. Cancer Res 64:7050-7057, 2004
3. Santos-Silva F, Fonseca A, Caffrey T, Carvalho F, Mesquita P, Reis C, Almeida R, David L, Hollingsworth M:
Thomsen-Friedenreich antigen expression in gastric carcinomas is associated with MUC1 mucin VNTR
polymorphism. Glycobiology, 2005 (in press)
21
Carcinogenesis
In 2005 the group aims to clarify the mechanisms of intestinal transdifferentiation of the gastric mucosa and to
define the Lewis and Secretor status, as well as MUC1 gene polymorphisms involved in Helicobacter pylori
infection (HP). The group will also build transfected gastric cell line models to study the role of Sialyl-Tn and
MUC2/Silalyl-Tn in HP adhesion and cancer cell behaviour.
To clarify the mechanisms of intestinal transdifferentiation (intestinal metaplasia – IM) of the gastric
mucosa. A)The group will explore further mechanisms of transactivation of the intestinal mucin MUC2, by
testing the hypothesis of methylation dependent transcription mediated by AP2. We identified two CpG sites
unmethylated in gastric lesions expressing MUC2, corresponding to putative AP2 binding sites. Luciferase
assays, using unmethylated and methylated constructs of MUC2 promoter will be used. B) We will study the
auto-regulatory CDX2/CDX2 loop by mapping candidate binding sites using ChIp. C) We will test the
hypothesis of transactivation of CDX2 by OCT-1, based on evidence in mouse colonic cells and on colocalization of both proteins in IM. D) We will test the hypothesis of transcriptional regulation of CDX2 by
SMADs, based on association of SMAD4 mutations with syndromes characterized by alterations in intestinal
differentiation/architecture and on preliminary data suggesting that SMAD4 regulates CDX2 promoter in vitro.
To build a model system to clarify the biologic role of mucin genes polymorphisms. The group
developed a model of gastric cancer cells transfected with MUC1 constructs with different VNRT lengths. We
will silence endogenous MUC1 expression using a vector system for inducible expression of short interfering
RNAs. We will explore the relevance of MUC1 expression and polymorphisms for the phenotype of gastric
cancer cells, motility, invasiveness and metastatic potential. We will also explore the relevance of MUC1
expression and polymorphisms for the adhesion of HP in the gastric cell model.
To clarify the role of Lewis and Secretor genotypes/phenotypes in gastritis and intestinal metaplasia. A)
The group will explore the possibility that FUT3 promoter methylation influences gene expression, enzyme
activity and Lewisa/sialyl Lewisa antigen expression. The hypothesis is based on preliminary data showing that
the demethylating agent 5-aza-2' deoxycytidine induces Lewisa antigen expression in gastric cell lines, and
might explain increased expression of this antigen in gastritis and IM. B) We will clarify the relevance of two
new mutations of the FUT2 gene in secretor status and in development of lesions of the gastric mucosa
induced by HP. We will test a large series of samples collected during 2004 in Estaleiros Navais de Viana do
Castelo.
To clarify the biologic role of sialylated antigens in intestinal metaplasia and gastric carcinoma. A) We
will use a gastric cell line stably transfected with ST6GalNAc-I to study if Sialyl-Tn inhibits interaction with HP.
This hypothesis is based on previous observations showing high Sialyl-Tn expression in IM, where HP is "autoeradicated". B) We will use recombinant sialyltransferases to design a MUC2 mucin glycopeptide containing
Sialyl-Tn to study its putative anti-adhesive properties for HP. This compound mimicks the naturally occurring
mucins in IM and will be tested in in vitro assays. C) Finally, we will test if Sialyl-Tn antigen facilitates the
process of gastric carcinoma invasion and metastisation. This hypothesis is based on data showing
associations between expression of Sialyl-Tn and aggressiveness and poor prognosis of the cases. We will use
in vitro assays to test cell adhesion, motility and invasion. We will also develop an in vivo model in nude mice.
The group will try to bring to a common frame the projects and activities on animal pathology leaded by Fátima
Gartner. Animal models are particularly interesting since glycosylation has common features in human and
animal models that can be very useful for comparative purposes.
The group will lounch a project of collaboration with Mozambique aiming to understand the so called "African
enigma", i.e. to try to clarify the reasons underlying the low incidence of gastric carcinoma and intestinal
metaplasia in a country with a high frequency of gastritis by Helicobacter pylori. In parallel to previous projects
of the group, the status of the gastric mucosa evaluated by biopsies, the genetic polymorphisms implicated in
cancer risk and dietary risk factors, will be evaluated.
The group will make efforts to launch the diagnostic kit for intestinal metaplasia in the market.
22
Genetics, Evolution and Pathology
Coordinator: Jorge Rocha, PhD
Pos-Doc student: Susana Seixas
Junior Research Students: Margarida Coelho and Rita Quental
Undergraduate student: Susana Pereira.
Research interests: The long term goals of our research are the characterization of the major evolutionary
forces that shaped the current human genetic diversity and the study of the implications of population history
and population structure in the patterns of health and disease. To achieve these goals our research focus both
on the study of specific populations and on the analysis of the evolutionary history of particular genes. At the
population level, we are particularly interested in the study of human groups derived from recent historical
events where admixture is an important component of population structure. At the gene level, we are interested
in characterizing the evolutionary history of loci associated with hereditary disease or that might have been
important in adaptation to environmental changes.
Projects: 1-“Analysis of the organization and distribution of DNA sequence variation in the serpin gene
subcluster located on chromosome 14q32.1.” (Pos-Doc project FCTBPD/12532/2003); 2- “Anthropogenetics of
São Tomé e Príncipe: a case study on human microevolution” (FCT-project; POCTI/42510/ANT/2001); Lactose
intolerance: study of prevalence and analysis of compatibility between the Breath Hidrogen Test (BH2) and
molecular diagnosis (Portuguese Society of Gastroenterology grant); 3- Bio-cultural adaptation: human
evolutionary responses to major changes in subsistence economy (FCT-project; POCTI/BIA-BDE/56654/2004)
Research activity: Our research during 2004 may be conveniently divided into two major lines of activity centered
on the analysis of populations and the study of specific genes: a) the characterization of the genetic structure of the
population of São Tomé; and b) the study of genetic variation in the in a Serine Protein Inhibitor (SERPIN)
subcluster and in the locus associated with human variation in lactose tolerance.
The study in São Tomé was made in colaboration with the Population Genetics group and Cíntia Alves (UPS) and
aimed at the identification of population subclusters based solely on the genetic information, independently of any
geographic or ethnolinguistic sampling criteria. Using a battery of anonymous markers we found an unprecendeted
level of genetic subdivision within this small 1000km2 island and were able to define three major sub-groups that
capture the major structure of the population. We are presently trying to assess to what extend these subgroups
are due to in situ genetic differentiation or reflect different regional origins of the slaves that peopled the island. In
any case, the structure that we have now identified provides a true genetic map of São Tomé, which can be used to
interpret the distribution of genetic variation that may be relevant to human disease. For example, we have already
gathered evidence that there is substantial variation in the distribution of genetic variants affording resistance to
malaria across the different sub-groups.
The study is of variation in the in a Serine Protein Inhibitor (SERPIN) subcluster that includes loci coding for α1antitrypsin (PI) and corticosteroid binding globulin (CBG) is part of Susana Seixas pos-doc, in collaboration with
Prof. Anna Di Rienzo from the Department of Human Genetics of the University of Chicago, USA. After a
systematic analysis of DNA sequence variation across several portions of this subcluster in two ethnically diverse
populations from Europe and Africa, we are currently analyzing the data. We are convinced to have found a
consistent signal of natural selection associated with one particular α1-antitrypsin variant in the African sample,
and will try to search for further relevant mutations that may provide a biological basis for the action of selective
mechanisms. This work illustrates the concept that evolutionary studies may provide relevant insights into the
understanding of the functional importance and medical relevance of human genetic variation. During 2004 we
have submitted to FCT a project based on Susana’s pos-doc plan, which is presently under evaluation.
Finally, the work on lactose tolerance illustrates our interest in loci that potentially play a role in the adaptation of
humans to environmental change. Lactose intolerance, the primitive condition in all mammals, limits the use of
fresh milk in adults due to the decline in lactase activity after the weaning phase. The prevalence of this trait in
humans is highly variable and is negatively correlated with milk drinking habits of different human populations. A
major hypothesis- the Gene Culture Coevolution Hypothesis- explains the distribution of lactose tolerance as the
result of the recent selection pressure caused by the advantages of milk drinking, but until recently this hypothesis
had been mostly based on geographic correlations. We have used microsatellite markers to assess the haplotype
variability associated with a candidate mutation for lactose tolerance in an heterogeneous set of African and
23
Southern European population samples. We used coalescent simulations to provide formal genetic evidence that
the mutation affording tolerance to lactose underwent a rapid increase in frequency due to selection around 12 000
years ago. Our results support the hypothesis that lactose tolerance reached its present distribution due to the
selective advantage of lifelong unrestricted use of milk in human societies that practiced dairyng after the Neolithic.
This work, which has been part of the Masters thesis of Margarida Coelho, has been submitted for publication and
is currently under review. The work is also part of a project that aims at understanding the way in which cultural and
environmental changes caused by modifications in subsistence patterns might have provided selective pressures
leading to the emergence of human genetic adaptation. The project, which was submitted to FCT in 2003 and has
been approved for financial support, is a collaboration with the groups of Profs Davide Pettener (University of
Bologne) and Giovanni Destro-Bisol (University of Rome1).
Genetics, Evolution and Pathology
In 2005 we plan to undertake the following activities:
a) To proceed with the research anchored in our previous projects, including the submission for publication of
major results from the analysis of genetic structure in São Tomé and on the characterization of genetic
diversity in the SERPIN subcluster.
b) To start the research activities of the project entitled “Bio-cultural adaptation: human evolutionary
responses to major changes in subsistence economy (FCT-project; POCTI/BIA-BDE/56654/2004)
c)
To submit PhD-plans for our Junior Research Students anchored on the projects already submitted and in
our major research goals
d) To deepen our collaboration with the Pedagogic University of Mozambique through the participation in a
project on Human Biological Diversity in Mozambique in collaboration with Dr. Sarah Blangero from the
Southwest Foundation for Biomedical Research (USA). This project will provide an opportunity to
collaborate in a multidisciplinary team with interests in biomedical research and to involve more our team
in the study of African populations, together with the groups of of Profs Davide Pettener (University of
Bologne) and Giovanni Destro-Bisol (University of Rome1).
e) To develop collaborations with other groups within IPATIMUP on the topic of the evaluation of genetic
susceptibility to cancer in admixed populations
24
Molecular Targets in Cancer
Coordinator: Ginesa García - Rostán, MD, PhD
Graduate students:
Severina Moreira (from 1st November 2003 till 30th August 2004)
Angela Margarida Amorin Costa (1st September 2004 till present)
Isabel Pereira Castro (1st October 2004 till present)
1.
Exordium to Aggressive Thyroid Tumour Histotypes : Poorly differentiated and Undifferentiated
Thyroid Carcinomas
Thyroid cancer is usually a disease with good prognosis, however, in up to 30% of the cases a drastic change
in tumour behaviour with progression from well-differentiated thyroid carcinoma, either papillary (PTC) or
follicular (FC) phenotype, to poorly differentiated (PDC) or undifferentiated carcinoma (UC) may occur.
Poorly differentiated carcinomas (PDC) have been identified as an additional thyroid
carcinoma histotype lying in an intermediate clinico-pathological position between well
differentiated and UC. Poorly differentiated carcinomas have a high propensity for local
recurrence and hematogenous spread and are associated with significant patient morbidity and
mortality. Frequently, PDCs are characterized by the presence of microscopic features of PTC or
FC indicating possible evolution from a well differentiated carcinoma. The molecular events that
predispose to such a malignant evolution remain unknown.
Undifferentiated carcinomas (UC) represent the most aggressive neoplasm, with evidence of
epithelial differentiation (keratin immunoreactivity), derived from the follicular epithelium. In a
recent review, it has been reported that as many as 50% of UCs show signs of pre-existing or
coexisting differentiated carcinomas or benign adenomas, supporting the possibility that UC may
arise as the consequence of a tumour progression pathway.
Most patients with UC die within 1 year after diagnosis and it is known that mortality rates associated with PDC
patients are also very high. Although, well differentiated thyroid cancers are effectively treated by the use of
radio-iodine, a huge proportion of PDC and all of the UC show an inability to take up radio-iodine as a result of
loss of expression of the iodine symporter. Currently, we lack of therapeutic tools to treat those uptake negative
thyroid cancers and there is evidence that this type of disease is increasing both in Europe and the US.
Development of novel therapeutic strategies that target growth pathways operating in more aggressive less
well-differentiated thyroid cancer are likely to be a priority for the effective treatment of thyroid cancer in the
future.
Molecular markers of more aggressive tumors would be, therefore, of great help in precisely defining
prognosis and therapeutical decision making.
2.
Molecular Genetics of Aggressive Thyroid Tumours
Accumulation of multiple independent mutations in oncogenes and tumour suppressor genes during the life of
an individual, appear to be required to conform the full malignant phenotype. Direct evidence for such
accumulation of mutant genes during thyroid carcinogenesis has been obtained from studies dealing with the
occurrence of mutations in proto-oncogenes, tumour suppressor genes and mismatch repair genes at
successive histopathological stages that mark the progression from normal follicular cells to a fully malignant
tumour type. The following multistep, genetic model of thyroid follicular cell tumorigenesis attempts to compile
all the somatic genetic events so far postulated to be involved in the development and progression of
sporadic thyroid neoplasms of follicular origin.
25
In bold appear those genes which are believed to play a more relevant role within particular steps of tumour
progression. The data are inferred from the different prevalence reported in the various tumour phenotypes.
So far only a reduced number of genes targeted by somatic alterations have been involved in the pathogenesis
and/or biological behaviour of aggressive thyroid tumors (PDC and UC):
•
Ras mutations are significantly associated with poorly differentiated and undifferentiated thyroid
tumors (55% and 52 % of the cases respectively). In fact, among PDC Ras activation constitutes
the most prevalent mutational event reported so far in the literature. Furthermore, Ras mutations
are associated with poor prognosis. Patients bearing Ras mutated tumors exhibit an increased
risk of distant metastasis at diagnosis or during the follow-up and poor survival, independently of
tumour differentiation and stage. [J. Clin Oncol 21(17): 3226-35, 2003, by G. García-Rostán, H.
Zhao, RL. Camp, M. Pollan, A. Herrero, J. Pardo, R. Wu, ML. Carcangiu, J. Costa, and G.
Tallini].
•
P53 is inactivated in most undifferentiated thyroid carcinomas and almost 50% of PDC.
[Prevalence reported in various series published in the literature and also from results not
shown by G. García-Rostán]
•
β-catenin is clearly dysregulated in aggressive thyroid tumours. Both, CTNNB1 exon 3
mutations and nuclear β-catenin localization are restricted to PDC (25% and 22% respectively)
and to UC (65% and 48% respectively). With respect to patient outcome, down-regulation of
membrane β-catenin expression, aberrant nuclear expression and exon 3 mutations are
associated with poor prognosis independent of conventional prognostic indicators for thyroid
26
cancer but not of tumour differentiation. [Am. J. Pathol, 158: 987-996, 2001, by G. GarcíaRostán, et al. and Cancer Research 59: 1811-1815, 1999, by G. García – Rostán et al.]
•
RET/PTC rearrangements are not common in PDC (13%). Despite being more frequent
among PDCs showing cytological or architectural features of papillary carcinoma, the
rearrangement do not represents a pivotal event in the progression from well-differentiated
papillary carcinoma to PDC. [J. Clin. Endocrinol. Metabol, 87: 370-79, 2002, by M. Santoro,
M. Papotti, G. Chiappetta, G. García-Rostán, M. Volante, Ch. Johnson, RL. Camp, F.
Pentimalli, C. Monaco, A. Herrero, ML. Carcangiu, A. Fusco, and G. Tallini].
•
Concerning the role of BRAF mutations in thyroid tumor dedifferentiation and progression no
un-doubtful relationship between BRAF mutation sites (activation loop, G-loop, or AKT sites)
and specific tumor histotypes with different degree of differentiation and/or biologic behaviour
has been consistently proved. Though activating mutations have been reported in
undifferentiated thyroid cancer cell lines and a small number of primary poorly differentiated
and undifferentiated thyroid carcinomas, the overall prevalence of oncogenic BRAF mutations
among aggressive thyroid tumor histotypes, their influence on papillary carcinoma
dedifferentiation, progression and/or metastatic potential, as well as their impact on patient´s
clinical course and survival remains unclear if not unknown. Further work is, thus, required in
order to quantify more precisely the strength of the association between BRAF activation and
aggressive thyroid tumor histotypes. In December 2003 we have started a project entitled
“Genomic and post-genomic in depth molecular characterization of the RAS/RAF/ERK signal
transduction pathway in aggressive thyroid tumors”. The major goals of the proposal are :
1.
Address the role of oncogenic BRAF mutations in the pathobiology of thyroid papillary
carcinomas and evaluate their prognostic impact
2.
Provide an in-depth molecular characterization of the RAS/RAF/ERK signalling
transduction pathway in the stepwise, time dependent progression of papillary
carcinoma towards the aggressive poorly differentiated and undifferentiated thyroid
carcinoma
Two of the most relevant milestones that we expect to achieve during the life of this project could be
summarized in :
•
Establish a set of genes that are modulated by RAS and BRAF in thyroid tumorigenesis using
microarray based expression profiling in different models (primary thyroid carcinomas, thyroid
cancer cell lines, rat thyroid epithelial cell lines and primary short term cultures).
•
Define the pharmacological profile of novel selective pharmaceutical compounds targeting
different molecules within the RAS/RAF/ERK signalling cascade and simultaneously gain insight
into the complex molecular mechanisms modulating drug sensitivity and resistance in patients
with prognostically unfavourable thyroid tumors
In summary, there are still many open questions regarding the pathobiology of aggressive thyroid tumors.
Despite over the last few years there has been an improvement in our knowledge of the molecular alterations
that underlie in PDC and UC, additional work needs to be done in order to uncover new genes and proteins
relevant for :
•
•
the initiation of aggressive thyroid tumors
the diagnosis of thyroid tumors histotypes with different degree of differentiation or
biologic behaviour (well-differentiated, poorly differentiated and undifferentiated thyroid
carcinomas). Particularly, the diagnosis of poorly differentiated thyroid carcinomas as a group of
neoplasms featuring a significant morbidity and mortality, for which there is not with the exception
of insular carcinoma, generally accepted morphological diagnostic criteria
27
•
•
•
the prognosis of thyroid carcinomas, anticipating the clinicians which papillary carcinomas or
follicular carcinomas will eventually progress into less differentiated neoplasms and which
tumours exhibit a greater risk of metastases
the prediction of therapy outcome
the development of novel, selective pharmaceutical compounds for treatment of thyroid
cancer patients harbouring primary tumours or metastasis resistant to I131
Recently developed post-genomic technologies will definitely help us in obtaining global molecular profiles
which should provide us with the ability of addressing fundamental questions such as whether we can detect
aggressive tumours as early as possible, whether a cancer is prone to acquire an angiogenic potential and to
metastasize,
which are the different prognostic outcomes of tumours otherwise morphologically
indistinguishable, whether we can predict different responses to therapy and circumvent resistance.
The analysis of the fluctuations of expression of a large pool of genes in cancer, might also provide a fresh
outlook on the most challenging problem of all in modern cancer research, i.e. the development of new
therapeutic approaches based on the molecular understanding of cancer
3.
Research activities in 2004
In year 2004 my situation as senior research scientist at the IPATIMUP improved a little bit in some aspects but
it is still far beyond the conditions I was told that I will have once in the IPATIMUP. In fact, two years and a half
after arriving to the institution I do not have even my own 4 meters of bench and the only laboratory help that I
have is two BICs that started to work with me in September and October 2004. One of them, is a substitution of
a previous BIC that was assigned to me 10 months after my arrival (November 2003). Besides the fact that
BICs do not usually have too much laboratory experience, the possibility of loosing them because they may
enter in a pre-doctoral program, as happened with the BIC that was working with me from November 2003 to
August 2004 makes the situation of the group difficult. On the other hand, is hard to submit grants when your
team is composed just of BICs. In 2004 I wanted to submit within the third call for proposals of the 6th
Framework Program a project on genomics and proteomics in thyroid cancer, based on an Expression of
Interest (EoI) that I sent in 2002 and a previous EU proposal (2001) in which I was the coordinator, but it was
not possible because I do not have the necessary team to do that type of project. I would need at least one
post-doc and a pre-doc or a technician plus some collaborations from other groups at the IPATIMUP. I am open
to collaborate with other groups at the IPATIMUP but I can not write an EU proposal in which 90% of the team
belongs to other groups. It is quite frustrating to reply the mails you get from groups or biotechs that have seen
your Expression of Interest on the web and are interested in participating in your proposal. Besides training,
supervise the work of the two BICs and do myself at extra-hours, weekends or vacations a great number of the
planned tasks, I need to deal myself with all the issues related with the daily ongoing lab work (order reagents,
look for the cheapest prices, fight with providers that never deliver the reagent or the equipment on the
estimated time. e.g. two months waiting for ethanol, or MDE………). On top of that, since January 2005 I am
surviving with a budget of 1100 euros per month.
Despite the situation described above it is difficult and sometimes I would say that very painful the RESEARCH
ACTIVITIES DEVELOPED ON MY GROUP AT 2004 INCLUDE :
Research grants submitted in 2004:
1.
Research proposal written for professor Nesland at the Radium Hospital in Norway. The project was
considered “excellent” by professor Nesland. Though he said he would be able to collaborate in laboratory
expenses I did not get the money yet.
2.
Research proposal written for the Norwegian Research Council. Waiting results
3.
Research proposal sent to the first call open by the FCT since I arrived to the IPATIMUP. The project
entitled “INACTIVATION OF MITOCHONDRIAL COMPLEX II TUMOR SUPPRESSOR GENES IN SPORADIC AND FAMILIAR
PARAGANGLIOMAS AND PHEOCROMOCYTOMAS” was approved last December. Is a one year project and the
budget is 20.000 euros. Reference Number : POCTI/SAU-OBS/61945/2004
28
4.
Research proposal sent to the first call open by the FCT since I arrived to the IPATIMUP. The project
entitled “GENOMIC AND POST-GENOMIC IN DEPTH MOLECULAR CHARACTERIZATION OF THE RAS/RAF/ERK AND
THE PI3K/AKT SIGNALLING TRANSDUCTION PATHWAYS IN AGGRESSIVE THYROID TUMORS” was approved this
week. Is a three years proposal and the budget is 93.100 euros. Reference Number: POCTI/SAUMMO/59607/2004.
Papers submitted for publication :
1.
Frequent mutation of PIK3CA in Anaplastic Thyroid Cancer. Ginesa García–Rostán, Angela M. Costa,
Isabel Pereira–Castro, Giuliana Salvatore, Severina Moreira, Radhames Hernandez, Agustin Herrero,
Alfredo Fusco, Jose Cameselle–Teijeiro, Massimo Santoro. Cancer Research
2.
Mitogenic effects of the up-regulation of MiniChromosome Maintenance proteins (MCM) in
anaplastic thyroid carcinoma. Teresa Guida, Giuliana Salvatore, Pinuccia Faviana, Riccardo Giannini,
Ginesa Garcia-Rostan, Livia Provitera, Fulvio Basolo, Alfredo Fusco, Francesca Carlomagno and Massimo
Santoro. Journal of Clinical Endocrinology and Metabolism.
3. CHK1 impairment and microsatellite instability in sporadic paragangliomas. Kathrin Riemann,
Simone Braun, Karl Sotlar, Susan Kupka, Carsten M Pusch, Carlos Suarez, Agustin Herrero Zapatero,
Jorge Lima, Markus Pfister Ginesa García-Rostán, Nikolaus Blin. Oncogene
4. High frequency of germline SDHB and SDHD mutations in sporadic head and neck paragangliomas
from northern Spain. Jorge Lima, Talia Feijao, Isabel Pereira-Castro, Angela Costa, Valdemar Maximo,
Agustin Herrero, Carlos Suarez, Manuel Sobrinho-Simoes, Ginesa Maria Garcia-Rostan. Journal Medical
Genetics. This work was developed mainly by Jorge Lima and Talia Feijao from the cell biology group and
myself.
5. Hürthle (oncocytic) cell tumours of thyroid: Etiopathogenesis, diagnosis and clinical significance.
Manuel Sobrinho-Simões, Valdemar Máximo, Inês Vieira de Castro, Elsa Fonseca, Paula Soares, Ginesa
Garcia-Rostan, Manuel Cardoso de Oliveira. International Journal of Surgical Pathology, 2005; 13 : 29-35.
In addition to the papers listed above in the first 6-7 months of 2005 we expect to be able to deal with 4
more papers as a result of the work developed in 2004 and the beginning of 2005. One of those papers will
be focused on the results of the genomic approach of the BRAF proposal described above. After summer
2005 we expect to be able to tackle the post-genomic step of that proposal.
Abstracts submitted to International Congress :
1.
96th Annual Meeting of the AACR. April 16-20, 2005, Anaheim, CA. “HIGH FREQUENCY OF GERMLINE
SDHB AND SDHD MUTATIONS IN SPORADIC HEAD AND NECK PARAGANGLIOMAS FROM NORTHERN SPAIN”. Jorge
Lima, Talia Feijao, Isabel Pereira-Castro, Angela Costa, Valdemar Maximo, Agustin Herrero, Carlos
Suarez, Manuel Sobrinho-Simoes, Ginesa Maria Garcia-Rostan. Accepted for Platform Presentation.
2.
7th Congress of the European Skull Base Society. 13th Congress of the German Society of Skull
Base Surgery. May 18-21, Fulda, Germany. “GERMLINE SDHB AND SDHD MUTATIONS IN APPARENTLY
SPORADIC HEAD AND NECK PARAGANGLIOMAS FROM SPAIN”. Jorge Lima, Talia Feijão, Isabel Castro, Angela
Costa, Valdemar Máximo, Agustín Herrero, Manuel Sobrinho-Simões, Ginesa García-Rostán, Carlos
Suárez.. Accepted for Poster presentation.
Local – National Meetings :
29
1.
2004 Porto Cancer Meeting. “ONCOGENIC BRAF MUTATIONS IN EXON-15 REPRESENT AN ALTERNATIVE
GENETIC EVENT TO RAS ACTIVATION IN THYROID TUMOR PROGRESSION/DE-DIFFERENTIATION”. Severina Moreira,
Agustin Herrero, Jose Cameselle, Ginesa Garcia Rostan. Poster presentation
I sincerely think that having the necessary help the results presented by my group in this annual report could be
better. Though I know that I should not do comparisons sometimes one can not avoid to do that type of
exercise. Comparing with some groups in the IPATIMUP in which there are 18 or 23 people with 6 or 8 PhD
probably the results are not bad. I apologize for the comparison but sometimes I feel a little abandon or as a
second category group leader
4. Relevant publications in the past few years are :
•
G. García-Rostán, H. Zhao, RL. Camp, M. Pollan, A. Herrero, J. Pardo, R. Wu, ML. Carcangiu, J. Costa,
G. Tallini ”RAS MUTATIONS ARE ASSOCIATED WITH AGGRESSIVE BEHAVIOR AND POOR
PROGNOSIS IN THYROID CANCER”. Journal of Clinical Oncology. 2003. Sep 1, 21(17): 3226-35.
Impact Factor JCR 2003: 10.86. Total Citations : 5
•
G. Moreno-Bueno, D. Hardisson, C. Sanchez, D. Sarrio, R. Cassia, G. García-Rostán, J. Prat, M.Guo, J.
Herman, X. Matias-Guiu, M. Esteller J. Palacios. ”ABNORMALITIES OF THE APC/β-catenin PATHWAY IN
ENDOMETRIAL CANCER”. Oncogene. 2002, 21, 7981-90. Impact Factor JCR 2003: 6.49. Total
Citations : 20
•
G. Chiappetta, P. Hui, L. Golden, BK. Kinder, G. García-Rostán, DA. Dillon, M. Santoro, A. Fusco, J.
Rosai, G. Tallini. “RET/PTC ONCOGENE ACTIVATION AND ASSESSMENT OF CLONALITY IN
THYROID NODULES WITH INCOMPLETE MORPHOLOGIC EVIDENCE OF PAPILLARY CARCINOMA :
A SEARCH FOR THE EARLY PRECURSORS OF PAPILLARY CANCER”. American Journal of
Pathology. Cellular and Molecular Biology of Disease. 2002, 160(6):2157-67. Impact Factor JCR
2003: 6.94 Total Citations : 15
•
M. Santoro, M. Papotti, G. Chiappetta, G. García-Rostán, M. Volante, Ch. Johnson, RL. Camp, F.
Pentimalli, C. Monaco, A. Herrero, ML. Carcangiu, A. Fusco, G. Tallini. “RET ACTIVATION AND
CLINICOPATHOLOGIC FEATURES IN POORLY DIFFERENTIATED THYROID TUMORS ”. Journal
Clinical Endocrinology and Metabolism. 2002, 87 : 370-379. Impact Factor JCR 2003: 5.87. Total
Citations : 14
•
G. García-Rostán, R. Camp, A. Herrero, ML. Carcangiu, DL. Rimm, G. Tallini. “β-CATENIN
DYSREGULATION IN THYROID NEOPLASMS : DOWNREGULATION, ABERRANT NUCLEAR
EXPRESSION AND CTNNB1 EXON 3 MUTATIONS ARE MARKERS FOR AGGRESSIVE TUMOR
PHENOTYPES AND POOR PROGNOSIS”. The American Journal of Pathology. Cellular and
Molecular Biology of Disease. 2001, 158 (3) : 987- 996. Impact Factor JCR 2003: 6.94. Total citiations
: 34
•
G. Tallini, G. García-Rostán, A. Herrero, D. Zelterman, G. Viale, S. Bosari, ML. Carcangiu.
“DOWNREGULATION OF p27 AND Ki67/ Mib 1 LABELING INDEX SUPPORT THE
CLASSIFICATION OF THYROID CARCINOMA INTO PROGNOSTICALLY RELEVANT CATEGORIES”.
The American Journal of Surgical Pathology. 1999, 23(6) : 678-685. Impact Factor JCR 2003 :
4.53. Total citations : 30
•
G. Tallini, A. Hsueh, S. Liu, G. García-Rostán, MR. Speicher, DC. Ward. “FREQUENT CHROMOSOMAL
DNA UNBALANCE IN THYROID ONCOCYTIC (HüRTHLE CELL) NEOPLASMS DETECTED BY
COMPARATIVE GENOMIC HYBRIDIZATION”. Laboratory Investigation. 1999, 79(5) : 547-555. Impact
Factor JCR 2003 : 4.41. Total citations : 25
•
G. García-Rostán, G. Tallini, A. Herrero, TG. D’Aquila, ML. Carcangiu, DL. Rimm. “FREQUENT
MUTATION AND NUCLEAR LOCALIZATION OF β-CATENIN IN ANAPLASTIC THYROID CARCINOMA”.
Cancer Research. 1999, 59April 15: 1811-1815. Impact Factor JCR 2003: 8.64. Total citations: 111
30
Population Genetics Group
Coordinator: António Amorim, PhD
Principal investigators: Maria João Prata, PhD; Leonor Gusmão, PhD; Luísa Pereira, PhD;
Post-Doc students: Luísa Azevedo, PhD
PhD students: Sandra Beleza, Sandra Martins, Alexandra Lopes, Ana Goios, Filipe Pereira, Elisabete Oliveira
MSc students: Maria Assunção Senra, Elisabete Martins
Graduate students: José Meirinhos, Sofia Quental, Iva Gomes, Helena Pereira, Raquel Matos, Pedro Soares
Historical 2004
The group aims at understanding the origin and evolution of (mainly) human genetic diversity and their
consequences and applications, both normal and pathological.
This requires the development of descriptive and analytical formal tools and techniques adequate to specific
genomic segments. The main results published last year in this area were
a) The construction of genetic databases for various human populations (Portugal, Spain, Basque Country,
Tunisia, Brazil, São Tomé e Príncipe, Cabinda, and Mozambique);
b) The establishment of coding rules for electronic databases (with an application to the canine mitochondrial
genome)
c) The development of predictive methods of sampling saturation evaluation in the context of population
genetic databases
As for the progresses in the unravelling of mechanisms of origin and evolution of genetic diversity, including
human demographic history, the most relevant contributions were
- the study of the gene pair PCDHX/Y (protocadherins), resulting from a hominid specific X/Y translocation,
in which it was possible to compare the evolutionary pathway of the same genomic sequence in distinct
chromosomes with different modes of transmission and expression (?), so that we were able to date the Y form
detected in Portugal to a time period coinciding with the end of the Last Glacial Maximum
- the detection of repetitive motifs in mtDNA and their connection with rearrangements of the molecule, the
development of a searching software downloadable from the IPATIMUP website, suitable for circular genomes,
and the characterisation of mtDNA lineages in patients with myopathies involving single deletions
- the correlation between the mutation rates heterogeneity in the mtDNA control region with secondary
structures as revealed by phylogenetic comparisons or through the free-energy minimisation method
- the inference of demographic expansion from Iberia to the rest of Europe after the Last Glacial Maximum
through the subtyping of the mtDNA haplogroup H
- the demonstration of cis-acting sequence elements determining the ornithine transcarbamylase (OTC)
locus hypermutability
- the unravelling of the molecular heterogeneity of the deficiency alleles at the thiopurine methyltransferase
(TPMT), their ethnic/geographic distribution spectrum and respective dates of origin
The development of applications is also worth of mention. Molecular diagnostics were significantly improved (as
for the detection of deletions at the OTC gene), the absence of association between mtDNA haplogroups and
oligozoospermia was demonstrated, and the use of uniparental markers to the characterisation and certification
of domestic animal species was carried further, in the context of the project: GENCERT (Male and female
lineages of domestic animals of economic interest: characterisation by non-invasive techniques), in a
consortium MCT-ADI (Agência de Inovação SA) with CaniSemen, SA.
1. Alves C, Gusmão L, Damasceno A, Soares B, Amorim A . Contribution for an African 1,6
autosomic STR database (AmpF/STR Identifiler and Powerplex 16 System) and a report on
genotypic variations, Forensic Sci Int 139: 201-205, 2004
2. Alves S, Rocha J, Amorim A, Prata MJ . Tracing the origin of the most common thiopurine 3,1
methyltransferase (TPMT) variants: preliminary data from the patterns of haplotypic association with
two CA repeats. Ann Hum Genet 68:313-23, 2004
3. Azevedo L, Climent C, Vilarinho L, Calafell F, Amorim A. Evidence for mutational cis-acting 6,3
factors affectingmutagenesis in the ornithine transcarbamylase gene. Hum Mutat 24:273, 2004.
4. Beleza S, Alves C, Reis F, Amorim A, Carracedo A, Gusmão L . 17 STR data (AmpF/STR 1,6
Identifiler and Powerplex 16 System) from Cabinda (Angola). Forensic Sci Int 141: 193-196, 2004
31
5. De Souza Goes AC, De Carvalho EF, Gomes I, DA Silva DA, Gil EH, Amorim A, Gusmão L . 2,0
Population and mutation analysis of 17 Y-STR loci from Rio de Janeiro (Brazil). Int J Legal Med.
2004 Nov 24; [Epub ahead of print]
6. García O, Martín P, Gusmão L, Albarrán C, Alonso S, De La Rua C, Flores C, Izagirre N, Peñas 1,6
R, Pérez JA, Uriarte I, Yurrebaso I, Alonso A . A Basque Country autochthonous population study of
11 Y-chromosome STR loci. Forensic Sci Int. 145:65-68, 2004
7. Lopes AM, Calafell F, Amorim A. Microsatellite Variation and Evolutionary History of PCDHX/Y 6,0
Gene Pair Within the Xq21.3/Yp11.2 Hominid-Specific Homology Block. Mol Biol Evol. 21: 20922101, 2004
8. Lopez AM, Alvarez S, Gusmao L, Alves C, Mesa MS, Albentosa A, Arribas G, Lopez R, Barrio 1,6
PA, Amorim A, Arroyo-Pardo E . Population data for 16 Y-chromosome STRs in four populations
from Pyrenees (Spain). Forensic Sci Int 140:125-9, 2004
9. Martín P, García-Hirschfeld J, García O, Gusmão L, García P, Albarrán C, Sancho M, Alonso A 1,6
. A Spanish population study of 17 Y-chromosome STR loci. Forensic Sci Int. 139:231-235, 2004
10. Pereira L, Cunha C, Amorim A . . Predicting sampling saturation of mtDNA haplotypes: an 2,0
application to an enlarged Portuguese database. Int J Legal Med 118:132-6, 2004
11. Pereira L, Van Asch B, Amorim A . Standardisation of nomenclature for dog mtDNA D-loop: a 1,6
prerequisite for launching a Canis familiaris database. Forensic Sci Int 141: 99-108. 2004
12. Schneider PM, Bender K, Mayr WR, Parson W, Hoste B, Decorte R, Cordonnier J, Vanek D, 1,6
Morling N, Karjalainen M, Carlotti CMP, Sabatier M, Hohoff C, Schmitter H, Pflug W, Wenzel R,
Patzelt D, Lessig R, Dobrowolski P, O'donnell G, Garafano L, Dobosz M, Knijff P, Mevag B,
Pawlowski R, Gusmão L, Vide MC, Alonso A, García O, Sanz P, Kihlgreen A, Bär W, Meier V,
Teyssier A, Coquoz R, Brandt C, Germann U, Gill P, Hallett J, Greenhalgh M . STR analysis of
artificially degraded DNA–results of a collaborative European exercise. Forensic Sci Int. 139:123134, 2004
13. Trovoada MJ, Pereira L, Gusmão L, Abade A, Amorim A, Prata MJ. Pattern of mtDNA Variation 3,1
in Three Populations from Sao Tome e Principe. Ann Hum Genet. 68: 40-54, 2004
Book Chapter
Gusmão L, Alves C (2004). Y chromosome STR typing. In Methods Mol. Biol. 297: 67-82. Humana Press.
ISI indexed Publications in Procceedings
Alves C, Amorim A, Pereira L, Gusmão L (2004) African STR data based on a newly developed tetraplex
fluorescent system (CD4, F13A01, FES and MBPB) Progress in Forensic Genetics 10: 133-135. Elsevier,
Amsterdam
Amorim A, Alves C, Pereira L, Gusmão L (2004) Genotyping inconsistencies and null alleles using
AmpFLSTR® Identifiler® and Powerplex® 16 kit. Progress in Forensic Genetics 10: 176-178. Elsevier,
Amsterdam
Beleza S, Quintans B, Salas A, Amorim A, Carracedo A, Gusmão L (2004) Microgeographic substructure of
NW Iberian Y chromosome STR haplotypes. Progress in Forensic Genetics 10: 296-298. Elsevier, Amsterdam
Beleza S, Salas A, Amorim A, Gusmão L, Carracedo A (2004) Microgeographic substructure of Northern
Portuguese mitochondrial DNA lineages: the female perspective of this region history. Progress in Forensic
Genetics 10: 386-388. Elsevier, Amsterdam
Gusmão L, Sánchez-Diz P, Alves C, Brión M, Beleza S, Pereira L, Blanco A, Prata MJ, Carracedo A, Amorim A
(2004) GATA C4 allele 17 as a marker for sub-Saharan origin of Y-chromosome lineages. Progress in Forensic
Genetics 10: 281-283. Elsevier, Amsterdam
Pereira L, Amorim A (2004) How much more should the Y-STR Haplotype Reference Database increase to
reach a pragmatic saturation level? Progress in Forensic Genetics 10: 88-90. Elsevier, Amsterdam
32
Pereira L, Amorim A (2004) Is selection at mtDNA really a major concern? Progress in Forensic Genetics 10:
109-111. Elsevier, Amsterdam
Pereira L, Richards M, Alonso A, Albarrán C, Garcia O, Macaulay V, Amorim A (2004) Subdividing mtDNA
haplogroup H based on coding-region polymorphisms––a study in Iberia. Progress in Forensic Genetics 10:
416-418. Elsevier, Amsterdam
Souto L, Gusmão L, Amorim A, Ferreira E, Côrte-Real F, Vieira DN, da Cruz e Silva DF (2004) POP-Gene
TIMOR: first forensic DNA marker study of East-Timor people. Progress in Forensic Genetics 10: 201-203.
Elsevier, Amsterdam
Trovoada MJ, Pereira L, Gusmão L, Abade A, Amorim A, Prata MJ (2004) Insights from pattern of mtDNA
variation into the genetic history of São Tomé e Príncipe. Progress in Forensic Genetics 10: 377-379. Elsevier,
Amsterdam
ISI indexed ABSTRACTS
Goios A, Pereira L, Nogueira C, Pereira C, Vilarinho L, Amorim A (2004) MtDNA in myopathies: absence of a
preferential haplotypic background for deletions and a search for inversions. Biochimica Biophysica ActaBioenergetics 1657: 25-26 Suppl. 1
Martins S, Gaspar C, Silveira I, Calafell F, Rouleau G, Coutinho P, Amorim A, Sequeiros J (2004) Portuguese
and non-Portuguese origins of Machado-Joseph disease (MJD): a worldwide haplotype study. Am J Hum
Genet 75: 230 Suppl. 1
Pereira L, Goios A, Nogueira C, Pereira C, Vilarinho L, Amorim A (2004) Directly repeated motifs occur more
often than expected in mtDNA. Biochimica Biophysica Acta-Bioenergetics 1657: 89 Suppl. 1
Invited ORAL PRESENTATIONS
Alves C: Y-STR mutations - Results from the 2003/2004 GEP-ISFG Collaborative Exercise. Promega's 6th
European Working Group Meeting, Barcelona, Espanha, 13-14/10/2004
Amorim A.Genética Forense. VI Jornadas de Biologia Aplicada, U Minho, Braga, 8-9/11/2004
Amorim A. Arqueogenética. II Fórum da Química. FCT/UNL. Monte da Caparica, 6-8/10/2004
Amorim A. Genética Forense. Curso de Mestrado em Medicina Legal. Coimbra, 18/10/2004
Amorim A. Population genetics and forensic sciences. University El Manar III. Tunis, Tunisia. 9/12/2004.
Calafell F, Lao O, Azevedo L, Sabater M, Soria JM, Amorim A, Bertranpetit J. The natural history of human
diseases. 8th Annual Meeting SPGH, 17-19 Nov 2004
Gusmão L: Y-STR’s allele frequency distributions within haplogroups. Workshop #21: Y-STR Analysis on
Forensic Casework. American Academy of Forensic Sciences 56th Meeting. Dallas, USA, 17/02/2004.
Gusmão L: Y-STRs allele distributions inside defined haplogroups. Meeting: New Technologies for human DNA
studies. International Academy of Legal Medicine. Strasbourg, France, 12-14/05/ 2004.
Gusmão L: Utilidad de los Marcadores Genéticos Moleculares en las pruebas de paternidad y en la
identificación human. Centro de Investigações Científicas y Tecnológicas (CICTE). Universidad de Cartagena.
Cartagena, Colômbia, 07/07/2004.
Gusmão L: Estrategias para la tipificación y secuenciación de marcadores genéticos. Instituto de
Investigaciones Inmunológicas. Universidad de Cartagena. Cartagena, Colômbia, 09/072004.
Gusmão L: Abordagens estatísticas e populacionais do cromossomo Y. I Jornada Forense Applied Biosystems:
Cromossomo Y-aplicações e abordagens estatísticas. São Paulo, Brazil, 07/11/2004.
Gusmão L: Marcadores Genéticos do Cromossoma Y em estudos de genética populacional e Investigação de
Parentescos. Laboratório de Diagnostico por DNA da Universidade do Estado do Rio de Janeiro, Brasil,
09/11/2004.
Gusmão L, Sánchez-Diz P, Martín P, Zarrabeitia MT, Whittle MR, Carvalho M, Bozzo WR, Farfán MJ, Prieto L,
Góes ACS, Palacio O, Pinheiro MF, Alonso C, Builes JJ, Borjas-Fajardo L, Di Lonardo AM, Corach D, VidalRioja L, Vieira da Silva CI, Carvalho EF, Alonso A, Carracedo A, Amorim A (2004) GEP-ISFG collaborative
studies on Y chromosome STR loci: methods, population data and mutation rates. IV International Forensic YUser Workshop. Haploid DNA markers in Forensic Genetics. November 18-20, Berlin, Germany.
Pereira, L. Aplicação da biotecnologia à ciência forense. Encontros de Biotecnologia. Angra do Heroismo. 1415/05/2004.
Pereira, L., Roewer, L., Amorim, A. Expectancy of Y-STR haplotype diversity parameters under different
population scenarios. IV International Forensic Y-User Workshop: “Haploid DNA markers in forensic genetics”.
Berlin, Germany. 18-20/11/2004.
Pereira, L. High-resolution mtDNA evidence for the late-glacial resettlement of Europe from an Iberian refugium.
University El Manar III. Tunis, Tunisia. 9/12/2004.
33
ORAL PRESENTATIONS
Goios, A., Meirinhos, J., Amorim, A., Pereira, L. Motivos repetidos no genoma mitocondrial: ocorrem mais
frequentemente que o esperado e são evolutivamente conservados. 8ª Reunião Anual da Sociedade
Portuguesa de Genética Humana, 18-19/11, Porto, Portugal
Pereira, L., Richards, M., Goios, A., Macaulay, V., Amorim, A. Repovoamento da Europa a partir de um refúgio
Ibérico (após o Máximo Glaciar) evidenciado por uma fina resolução do haplogrupo mitocondrial H.. 8ª Reunião
Anual da Sociedade Portuguesa de Genética Humana, 18-19/11,Porto, Portugal
Azevedo L, Soares P, Vilarinho L, Amorim A. Deficiência em ornitina transcarbamilase: diagnóstico molecular e
identificação de constitutições haplotípicas de risco. 8ª reunião SPGH, 17-19 Nov 2004
POSTERS presented to scientific meetings
Alves, C., Amorim, A., Gusmão, L., Pereira, L. (2004) Um novo alelo no locus D2S1338. IX Jornadas de
Genética Forense, GEP-ISFG. Manaus. Brasil. 02-04/06.
Alves S, Oliveira E, Quental S, Ferreira F, Costa V, Norton L, Sodré-Borges B, Amorim A, Prata MJ (2004)
Polimorfismos em genes da via metabólica dos folatos e susceptibilidade à leucemia linfoblástica aguda
infantil. 8ª Reunião Anual da Sociedade Portuguesa de Genética Humana. 17-19 Novembro 2004, Porto
Arroyo-Pardo E, Gusmão L, López A, Mesa MS, Amorim A (2004) Y-STR differentiation and substructuring in
Pyrenean populations. IV International Forensic Y-User Workshop. Haploid DNA markers in Forensic Genetics.
November 18-20, Berlin, Germany.
Beleza S, Amorim A, Carracedo A, Arroyo-Pardo E, Gusmão L (2004) Análise de STRs do cromossomoma Y
em Cabinda. IX Jornadas de Genética Forense. Reunião do GEP-ISFG (Grupo Espanhol e Português da
International Society for Forensic Genetics). Manaus 2-4/06, Brazil.
Sandra Beleza , Amorim Amorim , Angel Carracedo, Leonor Gusmão (2004), "17 Y-chromosome STR Loci
Variation in Cabinda (Angola) , "International Symposium on Human Identification", 15th edition, Phoenix,
Arizona, USA
Candon, A., Endicot, P., Torres, C., Rodrigues, C., Goios, A., Pereira, L., Amorim, A. (2004) Estudo do impacto
genético da islamização nas séries antropológicas do Rossio do Carmo, Mértola, Portugal (sécs. V-XIII).
Resultados preliminares. IV Congresso de Arqueologia Peninsular. Faro. Portugal. 14-19/09.
Goios, A., Morales, A., Rodrigues, C., Torres, C., Amorim, A., Pereira, L. (2004) Estudo da diversidade
genética feminina em Mértola (sudeste de Portugal) – os fenómenos de Islamização e/ou Mediterranização. IX
Jornadas de Genética Forense, GEP-ISFG. Manaus. Brasil. 02-04/06.
Goios, A., Morales, A., Rodrigues, C., Torres, C., Amorim, A., Pereira, L. (2004) O impacto genético da
islamização em Mértola. Resultados preliminares sobre a diversidade do DNA mitocondrial (mtDNA) an
população actual. IV Congresso de Arqueologia Peninsular. Faro. Portugal. 14-19/09.
Gomes I, Beleza S, Lopes A, Alves C, Amorim A, Gusmão L (2004) Distribuição haplotípica de 16 STRs do
cromossoma Y an população portuguesa. IX Jornadas de Genética Forense. Reunião do GEP-ISFG (Grupo
Espanhol e Português da International Society for Forensic Genetics). Manaus 2-4/06, Brazil.
Gomes I, Beleza S, Lopes A, Alves C, Amorim A, Gusmão L (2004) The Human Y chromosome: Distribution of
16 microsatellite markers in the Portuguese population. 15th International Symposium on Human Identification
Congress, Phoenix 4 -7/10, EUA.
Lopes A, Ross N, Crow TJ, Amorim A (2004), "Insights on X inactivation from X-Y transposed genes", 69th
Cold Spring Harbor Symposium on Quantitative Biology: Epigenetics", Nova Iorque, EUA.
Oliveira E, Alves S, Ferreira F, Costa V, Norton L, Sodré-Borges B, Amorim A, Prata MJ (2004) Folate
metabolic pathway and susceptibility to childhood acute lymphoblastic leukaemia in North Portugal. The role of
C677T and A1298C MTHFR Polymorphisms. CHILDREN with LEUKAEMIA -International Scientific
Conference, 6-10 September 2004 , London
Oliveira E, Alves S, Ferreira F, Norton L, Costa V, Amorim A, Prata MJ (2005) Implicações an terapia da
leucemia linfoblástica aguda infantil do polimorfismo genético da tiopurina S-metiltransferase. XLI Conferências
de Genética Médica. 3-4 Fevereiro 2005, Porto
Oliveira E, Alves S, Ferreira F, Costa V, Norton L, Sodré-Borges B, Amorim A, Prata MJ (2004) Polimorfismos
genéticos da MTHFR e susceptibilidade à leucemia linfoblástica aguda infantil em Portugal. 8ª Reunião Anual
da Sociedade Portuguesa de Genética Humana. 17-19 Novembro 2004, Porto
Pereira F, Pereira L, van Asch B, Amorim A (2004) Mitochondrial DNA diversity of Portuguese goat breeds.
Portugaliae Genetica, 7th edition, Porto, Portugal.
Pereira F, Pereira L, van Asch B, Amorim A (2004), “South Portugal sheep breeds: mtDNA genetic analysis”,
Portugaliae Genetica, 7th edition, Porto, Portugal.
34
Pereira F, Soares P, Amorim A, Pereira L (2004), "Análise das estruturas secundárias da região de controlo do
DNA mitocondrial" 8.ª Reunião Anual da Sociedade Portuguesa de Genética Humana, Porto, Portugal.
Quental S, Alves S, Quental S, Ferreira F, Costa V, Norton L, Amorim A, Prata MJ (2004) Polimorfismo
genético da metionina sintetase reductase e susceptibilidade à leucemia linfoblástica aguda infantil em
Portugal. 8ª Reunião Anual da Sociedade Portuguesa de Genética Humana. 17-19 Novembro 2004, Porto
Souto L, Ferreira E, Gusmão L, Amorim A, Côrte-Real F, Vieira DN (2004) Luís Souto Aplicação de
Marcadores Polimórficos do Cromossoma Y com Interesse Médico-legal no estudo da população de TimorLeste. IV Congresso de Investigação em Medicina: Pedagogia e Ciência. Faculdade de Medicina de Coimbra
17-18/11, Portugal.
Souza Góes AC, Fagundes de Carvalho E, Gomes I, Aparecida da Silva D, Fonseca Gil EH, Amorim A,
Gusmão L (2004) Diversidade haplotípica de 16 STRs do cromossoma Y an população do Rio de Janeiro
(Brasil). IX Jornadas de Genética Forense. Reunião do GEP-ISFG (Grupo Espanhol e Português da
International Society for Forensic Genetics). Manaus 2-4/06, Brazil.
van Asch B, Pereira L, Pereira F, Santa-Rita P, Lima M, Amorim A (2004) Mitochondrial DNA lineages of four
Portuguese autochthonous dog breeds: diversity and phylogeography. Portugaliae Genetica, 7th edition, Porto,
Portugal.
Prizes
Young Researcher SPGH (Sociedade Portuguesa de Genética Humana) 2004 – Honorable Mention as 1st.
author of the poster: “Análise das estruturas secundárias da região de controlo do DNA mitocondrial” (Filipe
Pereira, Pedro Soares, António Amorim, Luísa Pereira) presented at the 8th. Reunião Anual da Sociedade
Portuguesa de Genética Humana.
Best Poster XLI Conferências de Genética Instituto Genética Médica (2004) - Oliveira E; Alves S; Quental S,
Ferreira F; Norton L; Costa V; Amorim A; Prata MJ.;Implicações an terapia da leucemia linfoblástica aguda
infantil do polimorfismo genético da tiopurina S-metiltransferase
PhDs
- Azevedo L. (FCUP, supervisor: A Amorim; 02/02/2004)
- Trovoada MJ (FCTUC, co-supervisor MJ Prata, 14/12/2004)
- Beleza S. Phylogenetic and demographic history of two human populations revealed by the analysis of two
non-recombining segments of the genome: Y-chromosome and mitochondrial DNA (U Santiago de
Compostela; supervisor: L Gusmão); submitted.
- Pereira F “Development of uniparentally transmitted genetic markers for the characterization of male and
female gene pools of Portuguese small ruminants autochthonous breeds” Faculty of Sciences, University
of Porto, IPATIMUP, and Department of Genetics, Smurfit Institute, Trinity College, Dublin 2, Ireland.
Fundação para a Ciência e Tecnologia (SFRH/BD/19585/2004). Since October 2004
- Oliveira E “Study of Genetic Polimorphisms in Pediatric Acute Lymphoblastic Leukemia - Pharmacogenetic
Role in the Treatment and Relationship with susceptibility to the leukemogenic process” Faculty of
Sciences, University of Porto, IPATIMUP, and School of Medicine , Washington University in St. Louis.
Fundação para a Ciência e Tecnologia (SFRH/BD/17124/2004). Since November 2004.
- Goios A “Dynamics of the mitochondrial genome – the moving boundary between normal and pathogenic
diversity” Faculty of Sciences, University of Porto, IPATIMUP and Department of Statistics, University of
Glasgow. Fundação para a Ciência e Tecnologia (SFRH/BD/16518/2004). Since November 2004.
- Pedro Soares obtained a Marie Curie Early Stage Training in Advanced Genetics Analysis Grant, for his
PhD at Leeds Univ.
- Iva Gomes started her PhD in the Univ. Santiago de Compostela
Scientific Seminars
- (Starting to) SNP at admixture. Vincent Macaulay (Dept. Statistics, U. Glasgow) 26/05/2004
- mtDNA and the origins of Orang Asli. Martin Richards (Schools of Biology and Computing, U. Leeds)
26/05/2004
Visiting researchers at IPATIMUP
Lotfi Cherni (U Tunis, Tunisia) 19/07-06/08
Eduardo Arroyo-Pardo (U Complutense, Madrid) 18/08-18/09
35
Elizeu Fagundes de Carvalho (Laboratório de Diagnostico por DNA da Universidade do Estado do Rio de
Janeiro), 21-26/04 e 22-24/11
Andréa Carla de Souza Góes (Laboratório de Diagnostico por DNA da Universidade do Estado do Rio de
Janeiro), 14/04-01/05
Visits / Courses Abroad
A Lopes (Dep. Psychiatry, U. Oxford, 7/11-21/12)
A Amorim, L Pereira (Univ Tunis, 06-12/12/2004; included in the Project GRICES (Proc 4.1.5 Tunisia)
S Martins (45th Annual Short Course in Medical and Experimental Mammalian Genetics, The Jackson Lab, Bar
Harbor, EUA, 18-30/07)
C Alves (Instituto Nacional de Toxicologia e Ciências Forenses de Madrid, Julho 2004)
Organization of Scientific Meetings
Portugaliae Genetica 7th. Edition HUMANS AND OTHER DOMESTICATES
Annual Meetion of SPGH (Sociedade Portuguesa de Genética Humana)
National Projects
approved during 2004
Estudo da heterogeneidade fenotípica patológica usando a proteína ornitina transcarbamilase (OTC) como
modelo. PI: G Suriano (PI), with L Azevedo
Estudo evolutivo e epidemiológico de focos de anemias hereditárias em Portugal – POCTI/ANT/57037/2004.
PI: MJ Prata
Etnias e genética an fronteira da rota de migração Bantu: o caso dos Karamajong. PI: L Gusmão
submitted, not evaluated yet
Origins and dispersals of Machado-Joseph Disease (MJD) mutations. A molecular and evolutionary approach
to the heterogeneity of a late onset hereditary ataxia. POCTI/SAU-MMO/58854/2004. PI: A Amorim
“Infertilidade humana, sucesso reprodutivo e transmissão do DNA mitocondrial” POCTI/MMO/59474/2004. PI:
L Pereira
Análise estrutural e molecular das regiões pseudoautossómicas humanas. POCTI/BIA-PRO/56584/2004. PI: L
Azevedo
International Projects
EC Submitted (PI: A Amorim): Selection and development of an assay of polymorphisms as a standard
genomic control in association studies. (Acronym: IND-TAG)
International Jurys
A Amorim: PhD jury U Pompeu Fabra, Barcelona (Stéphanie Plaza, 02/04/2004)
A Amorim: 26/03/2004 PhD jury, U Santiago de Compostela (Paula Sánchez-Diz, 26/03/2004)
Others
Coordination of GABBA PhD Program (A Amorim)
Organisation of the course "Técnicas Laboratoriais Moleculares", attended (and payed) by Carla Alice Afonso
and Paula Alexandra Ramos Lopes dos Santos, employees from Segalab, SA, 3-7 de Maio de 2004.
2005 Plan
The most important research developments we aim in the area relating to population genetics and databases
are: (a) the effects of the Bantu expansion in reshaping the African genetic landscape; (b) the assessment of
the demographic exchanges among Europeans and between Europe and North Africa; (c) the genetic affinities
of the populations of the islands of East Timor, Azores, and of the Gulf of Guinea; (d) forensic and diagnostic
applications and quality control.
As for the study of domestic species we intend (a) to join the international consortium aiming at the mouse
genome sequencing and to characterise the mitochondrial lineages of the lab strains; (b) to complete the study
of Portuguese dog, sheep and goat autochthonous breeds.
Concerning the evolutionary studies we plan (a) to finish the research of the PCDHX/Y gene pair with the study
of methylation levels of promoter regions in each of the heterosomes and (b) to clarify the molecular
mechanism responsible by the presence of an OTC pathogenic substitution in humans as wild type in
chimpanzees
36
The lab mouse model will be used for the research of mtDNA mutagenesis, which will be also studied in
humans in the context their pathogenic consequences and fertility impact.
Using the model of the MJD (Machado-Joseph disease) locus we will study (in normal and affected
populations) the mutation mechanisms underlying the DNA repetitive tract expansion.
During this year we will organise the scientific meetings:
8th. Portugaliae Genetica
21st. Congress of the International Society for Forensic Genetics.
(in collaboration with Portuguese National Institute of Legal Medicine (INML)
2005
Alonso A, Alves C, Suarez-Mier MP, Albarran C, Pereira L, Fernandez DE Simon L, Martin P, Garcia O,
Gusmão L, Sancho M, Amorim A. Mitochondrial DNA haplotyping revealed the presence of mixed up benign
and neoplastic tissue sections from two individuals on the same prostatic biopsy slide. J Clin Pathol 58(1): 83-6,
2005
Alves C, Gusmão L, Lopez-Parra AM, Mesa MS, Amorim A, Arroyo-Pardo E. STR allelic frequencies for an
African population sample (Equatorial Guinea) using AmpFlSTR Identifiler and Powerplex 16 kits. Forensic Sci
Int. 148:239-42, 2005
Arroyo-Pardo E, Gusmão L, López-Parra AM, Baeza C, Mesa MS, Amorim A. Genetic variability of 16 Ychromosome STRs in a sample from Equatorial Guinea (Central Africa). Forensic Sci Int. (in press).
Cherni L, Loueslati Yaacoubi B, Pereira L, Alves C, Khodjet El Kill H, Ben Ammar EL, Gaaied A, Amorim A.
Data for 15 autosomal STR markers (Powerplex 16 System) from two Tunisian populations: Kesra (Berber) and
Zriba (Arab). Forensic Sci Int 147(1):101-6, 2005
Diederiche M, Martín P, Amorim A, Corte-Real F, Gusmão L (2005) A case of double alleles at three Y-STR
loci: forensic implications. Int J Legal Med. (in press).
Montiel R, Bettencourt C, Silva C, Santos C, Prata MJ, Lima M. Analysis of Y-chromosome Variability and its
Comparison with mtDNA Variability Reveals Different Demographic Histories Between Islands in the Azores
Archipelago (Portugal). Ann Hum Genet 69:1–10, 2005
Pereira L, Richards M, Goios A, Alonso A, Albarran C, Garcia O, Behar Dm, Golge M, Hatina J, Al-Gazali L,
Bradley Dg, Macaulay V, Amorim A . High-resolution mtDNA evidence for the late-glacial resettlement of
Europe from an Iberian refugium. Genome Res. 15: 19-24, 2005
Salas A, Prieto L, Montesino M, Albarrán C, Arroyo E, Paredes-Herrera MR, Di Lonardo AM, Doutremepuich C,
Fernández-Fernández L, González De La Veja A, Alves C, López CM, López-Soto M, Lorente JA, Picornell A,
Espinheira RM, Hernández A, Palácio AM, Espinoza M, Yunis JJ, Pérez-Lezaun A, Pestano JJ, Carril JC,
Corach D, Vide MC, Álvarez-Iglesias V, Pinheiro MF, Whittle MR, Brehm A, Gómez J : Mitochondrial DNA
error prophylaxis: assessing the causes of errors in the GEP’02–03 proficiency testing trial. Forensic Sci Int,
148(2-3): 191-198, 2005.
Souto L, Alves C, Gusmão L, Ferreira E, Amorim A, Côrte-Real F, Vieira DN (2005) Population analysis of 15
STRs on a sample from East-Timor. Forensic Sci Int. (in press).
Beleza S., Gusmão L., Amorim A., Carracedo A., Salas A. The genetic legacy of western Bantu migrations.
Hum Genet (in press)
Cherni, L., Loueslati. B.Y., Pereira, L., Ennafaâ, H., Amorim, A., El Gaaied, A.B.A. (2005) Female gene pools of
Berber and Arab neighbouring communities in Central Tunisia – the micro-structure of mtDNA variation in North
Africa. Hum. Biol. 77:61-70.
Amorim, A., Pereira, L. (2005) Pros and cons in the use of SNPs in forensic kinship investigation: a
comparative analysis with STRs. Forensic Sci. Int. (in press).
Pereira, L., Gonçalves, J., Goios, A., Rocha, T., Amorim, A. Human mtDNA haplogroups and reduced male
fertility: real association or hidden population sub-structuring. Int. J. Andrology (in press).
Pereira, L., Cunha, C., Amorim, A. (2005) The African female heritage in Iberia – a reassessment of the present
distribution of mtDNA lineages. Hum. Biol. (in press).
Cherni, L., Pereira, L., Goios, A., Yacoubi-Loueslati, B., Khodjet El Khil, H., Gomes, I., Gusmão, L., Alves, C.,
Slama, A., Amorim, A., Benammar El Gaaied, A. Y-chromosomal STR haplotypes in three ethnic groups and
one cosmopolitan population from Tunisia. Forensic Sci. Int. (in press).
Souto L, Gusmão L, Ferreira E, Amorim A, Côrte-Real F, Vieira DN (2005) Y-chromosome STR haplotypes in
East Timor: Forensic evaluation and population data. Forensic Sci Int. (in press).
37
Tumour Evolution and Development
Coordinator: Luís Teixeira, PhD
Graduate students:
Ângela Costa, BSc – BIC
Elisabete Ricardo – Undergraduate student (through September 2004)
Lara Henriques – Undergraduate student (through October 2004)
Marta Novais, BSc – BIC
Objectives/Goals of the research activity
Our main long term goal is to understand the molecular mechanisms underlying tumor evolution. Rather than
focusing on na organ or tissue-specific type of tumor, we will pursue this goal by using different models to try to
answer specific fundamental questions within that broad goal. At this early stage, this was translated into
focusing on problems with which we had previous experience and felt we have the technical and financial
means to tackle: a) the regulation of TCF4, which is a key player in intestinal tumorigenesis; b) the role played
by 53 mutations in chromosomal instability in tumors.
Background and major achievements during 2004
2004 was a transition year for the lab. On the one hand, there was still a lot of “set-up” effort, with new
equipment and techniques being slowly added, and various projects entering exploratory or early
implementation stages. On the other hand, we made significant progress in the execution of one our FCTfinanced projects, “Identification of TCF4-interacting proteins”, which is now approaching completion.
Our interest in TCF4 stems from the PI’s previous experience in colorectal cancer, a tumor type initiated, in a
large majority of cases, by inactivation of (both copies of) the tumor suppressor gene APC. This inactivation
results in an abnormal increase in the cellular levels of a dimeric transcription factor formed by beta-catenin and
members of the TCF-LEF transcription factor family, in particular TCF4. TCF4 has also been shown to be a key
player in the differentiation of mouse gut epithelial cells, a role consistent with the fact that beta-catenin is an
effector of the "Wnt-Wingless" signaling pathway, whose importance in a number of developmental processes
in multiple animal species is well established.
Our strategy to study the regulation of TCF4 involves the use of a yeast two-hybrid system. During 2004 we
completed the screen of a human fetal brain cDNA library. Due to the large number of potential candidates
obtained, we were forced to resort to a secondary screen, after which that number was reduced to 179. The
yeast phase of the project was completed with the combination of insert-sequencing and individual re-testing of
cDNAs, yielding 4 novel potential TCF4-interacting proteins (including two uncharacterized proteins, known only
from cDNA databases). The corresponding cDNAs have been transferred to mammalian expression vectors
(adapted by us) and are now being tested by co-immunoprecipitation and a mammalian two-hybrid system.
Partial results have been reported to FCT and presented at the XIV National Meeting of Biochemistry. Two
undergraduate “theses” were produced as a result of this project; one on the identification of potential TCF4
interactors, another on human separin interactors. Additionally, we have been working on improvements of the
two-hybrid system that should facilitate both the elimination of false-positives in yeast and testing of bonafide
candidates in mammalian cells, thus improving the system’s efficiency.
38
4. ACTIVIDADES DA UNIDADE EDUCAÇÃO CONTÍNUA E DIFUSÃO CIENTÍFICA
39
A. Acreditação do IPATIMUP como entidade formadora
O IPATIMUP submeteu, com sucesso, uma candidatura ao Conselho Científico-Pedagógico da Formação
Contínua. É doravante uma entidade formadora acreditada.
B. Projectos e Programas
1-Programa “Ciência Viva em Férias”
O Programa “Ciência Viva em Férias” facultou a 14 alunos (da região Norte, Centro e Sul do País) um estágio
de duas semanas, que decorreu entre os meses de Junho a Setembro, 2004.
2- Programa “Viver uma Escola Diferente”
Protocolo com o Pelouro da Educação da Câmara Municipal do Porto. Programa “Viver uma Escola Diferente”.
Escolas (4) do primeiro ciclo do ensino básico. Janeiro 2004.
3- Projecto “Promoção e Internacionalização das Ciências da Vida no Norte de Portugal”
Continuação do Projecto “Promoção e Internacionalização das Ciências da Vida no Norte de Portugal”, que
teve inicio a 1 de Outubro de 2003. Tendo como entidade financiadora o CCDR-N (Comissão de Coordenação
e Desenvolvimento Regional -Norte) – Programa Norte. E Parceria Científica com o IBMC e o INEB.
4- Programa “Autolaboratório – Da Célula ao ADN”
Iniciativa de promoção do ensino experimental das ciências na sala de aula, incluída no projecto 3. Visita a 36
escolas da região Norte do País, em carro próprio adaptado em laboratório ambulante.
5- Projecto “A Magia da Ciência”
Preparação do projecto “A magia da Ciência” submetido ao concurso promovido pela FCT – “Atribuição de
Financiamento para projectos de divulgação da cultura científica e tecnológica” - Programa POCTI do III
Quadro comunitário de apoio - Medida 3.1.
C- Conferências, Colóquios e Palestras
1- Conferencia do Equinócio – “Saber e Cidadania”
VIII conferencia do Equinócio intitulada “Saber e Cidadania”, realizada a 18 de Outubro de 2004. Com a
coordenação da Doutora Maria de Sousa e o conferencista Dr. Mário Soares.
2- 1º Ciclo De Colóquios Sobre A Medicina Preventiva Do Cancro
Ciclo de 7 Colóquios organizado pela Fundação Calouste Gulbenkian e o IPATIMUP com a colaboração de
médicos e cientistas das Faculdades de Medicina e de Ciências da Nutrição e Alimentação das Universidades
do Porto, Coimbra e Lisboa, assim como dos Centros do Instituto Português de Oncologia do Porto e Lisboa e
dos Hospitais Universitários de Coimbra, Lisboa e Porto. Tema: Prevenção do Cancro, na lógica da
compreensão e transmissão dos conhecimentos actuais sobre a génese, desenvolvimento e potencial
prevenção das doenças cancerosas. População-alvo: Professores de Biologia e Físico-Química do Ensino
Secundário (400-700 de todo o País).
Colóquios realizados em Lisboa no Auditório da Fundação Gulbenkian de 1 de Março a 3 de Maio de 2004.
3- 2º Ciclo De Colóquios Sobre A Medicina Preventiva Do Cancro
Ciclo de 7 Colóquios organizado pela Fundação Calouste Gulbenkian e o IPATIMUP, com a apoio da
Fundação Serralves, e com a colaboração de médicos e cientistas das Faculdades de Medicina e de Ciências
da Nutrição e Alimentação das Universidades do Porto, Coimbra e Lisboa, assim como dos Centros do
Instituto Português de Oncologia do Porto e Lisboa e dos Hospitais Universitários de Coimbra, Lisboa e Porto.
Repetição do Ciclo 2. na cidade do Porto, no Auditório do Museu de Serralves de 19 de Novembro a 6 de
Dezembro de 2004
4- Colóquio “Porto de Ciência”
Participação no Porto de Ciência, com os Profs. Doutores Manuel Sobrinho- Simões e Rui Mota Cardoso. A 15
de Janeiro de 2004 no café Guarany.
40
5- Palestra realizada na Escola Secundaria Monte da Ola – Viana do Castelo pelos Drs. Vítor Gradissimo e
Ana Maria Ferreira intitulada “Cancro gástrico e a alimentação”. 1/4/2004.
6- Palestra realizada na Escola Secundária Condes Resende – Canelas pela Doutora Paula Soares e Drº Luís
Cirnes com o tema “Cancro do pulmão”, inserida na semana cultural. 23/04/2004.
7- Palestra realizada na Escola primária Perpétuo0 Socorro – Porto pela Doutora Paula Soares e Drº Luís
Cirnes intitulada “O alfabeto da genética”. 27/4/2004.
8- Palestra realizada na Escola Secundária de Canelas – Vila Nova de Gaia, pela Prof. Doutora Leonor David
com o tema “Genética e o cancro”, inserida no dia Nacional da Ciência. 24/11/2004
D- Exposições
1- Exposição dos trabalhos realizados ao longo do ano lectivo 2003-2004, pelas escolas do primeiro ciclo do
ensino básico. Inseridas no protocolo com o Pelouro da Educação da Câmara Municipal do Porto. Programa
“Viver uma Escola Diferente”. 3 a 5 de Junho 2004.
2- Participação no dia da Universidade do Porto, que decorreu no Pavilhão Rosa Mota – Porto. Conjunto de
experiências sobre os temas célula e ADN. Contou com a colaboração de: Ana Ferreira, Rita Mateus, Cátia
Moutinho, Gonçalo Regalo, Victor Gradíssimo, Ana Costa, Tiago Botelho, Luís Cirnes e Sílvia Carvalho. Tendo
também a colaboração de Luísa Ayres. Nos dias 18 a 21 de Março de 2004
3- Exposição do EpistemoZóide de Fabíola Valença no IPATIMUP a 2/6/2004
D- Outros
1- No dia Nacional da Cultura Cientifica, o IPATIMUP esteve de portas abertas para receber os alunos que
frequentaram o “Ciência Viva em Férias”. Intitulado -“ Trás um amigo também”. 24/11/2004
2- Apoio ao Laboratório de Ciência no Museu dos Transportes e Comunicações relativamente à exposição
permanente aí existente.
Porto, 14 de Março de 2005
Rui Mota Cardoso
41
5.
SERVIÇO À COMUNIDADE
5a. UPS
5b. UPSi
5c. UPSs
42
5a) UNIDADE DE PRESTAÇÃO DE SERVIÇOS (UPS)
Introdução
De forma a cumprir um dos objectivos do CIBO/IPATIMUP para 2004: “continuar a proceder à preparação para
o processo de acreditação da UPS via Colégio Americano de Patologistas (CAP), graças ao apoio do
Programa Operacional de Saúde (Saúde XXI)”, uma das prioridades neste ano foi implementar correctamente
o Sistema de Gestão pela Qualidade (SGQ) que definiu no transcorrer de 2003. Procuramos envolver todos os
colaboradores da Unidade na definição e execução dos procedimentos reguladores e actividades relacionadas
a higiene e segurança no trabalho. A partir de Abril de 2004 implementamos um novo software operacional
(SISLAB) que garante melhorias significativas nas actividades de controlo de qualidade, sendo desactivado o
programa anteriormente utilizado. Apesar da diminuição do número total de exames realizados, facto devido a
maior concentração em exames que tenham maior relação custo-benefício, não houve diminuição da
facturação total. A Unidade mesmo com a enorme concentração de esforços de preparação da acreditação
continuou a prestar serviços gratuitos de consultadoria para vários países (ver item # 5), bem como manteve o
nível de publicações com material obtido na sua rotina. Como vem sucedendo há vários anos, continuámos a
actuar como um centro de formação profissional pós-graduado, tendo recebido em 2004, 7 patologistas e 2
técnicos de diferentes países.
1.
Recrutamento de pessoal:
•
•
•
Em Julho de 2004, rescindimos contrato com a administrativa Cristina Brás.
Em Julho de 2004 contratamos, em regime de estágio, a técnica Regina Pinto.
Em Setembro de 2004 contratamos, em regime de contrato de estágio, a administrativa Cecília Seabra,
pelo período de 6 meses.
Em Outubro de 2004, contratamos, em regime de recibos verdes, a médica patologista Teresa Iscar
Gálan.
•
2. Aquisição de Equipamento:
Com a finalidade de equipar o laboratório de rotina diagnóstica, no âmbito do Programa de Acreditação da
Unidade via CAP com o Apoio do Programa Saúde XXI, foram adquiridos os seguintes equipamentos:
Aparelho/Empresa – Modelo
Preço s/ IVA
IVA (19%)
Programa do CAP*
2524,82 €
s/ IVA
Sistema Arquivo Istoglass + Istobloc**
1813,20 €
344,51 €
Combinado Siemens**
922,69 €
175,31 €
Armário duplo c/ ventilação forçada**
740,00 €
140,60 €
(*) O Projecto Saúde XXI comparticipa com 75% do total.
(**) Adquirido integralmente com recursos da Unidade.
3. Estágios, visitas de curta duração e treino de internos:
43
•
Estágios
Nome
Período
Tipo de Estágio
Teresa Iscar Gálan, Interna de Anatomia
02/01/2004 a
Anatomia Patológica,
Patológica, Hospital de La Princesa, Madrid,
31/03/2004
Citopatologia e
Espanha
Patologia Molecular
Marcus Milhone, Interno de Anatomia 29/10/03 a 31/10/04 Patologia da mama e
Patológica Faculdade de Medicina da
da tireóide
Universidade Federal do Ceará, Fortaleza,
Ceará
Regina Pinto, estudante da Licenciatura de 01/01/04 a 30/06/04 Técnicas laboratoriais
Anatomia
Patológica,
Citológica
e
Tanatológica, Instituto Politécnico de Saúde do
Norte, Paredes, Portugal
Jesus Alberto Veiga Barreira, Interno de 01/02/03 a 17/04/03 Patologia cirúrgica e
Patologia, Complexo Hospitalario Juan
citopatologia
Canalejo, La Coruña, Espanha
Rozany Mucha Dufloth, Especialista em 01/07/04 a 31/08/04
Patologia da Mama
Anatomia Patológica, Universidade Estadual
de Campinas, Brasil
Ricardo Camillo de Almeida, Interno de
06/09/2004 a
Patologia cirúrgica e
Anatomia Patológica, Faculdade de Medicina
1/11/2004
citopatologia
da Universidade de São Paulo, SP, Brasil
José Ajax Nogueira Queiroz, Especialista,
01/12/2004 a
Citometria de Fluxo
Faculdade de Medicina da Universidade
20/01/2005
Federal do Ceará, Fortaleza, Ceará
Paulo Roberto Carvalho de Almeida, 25/10/04 a Fevereiro
Patologia gástrica
Especialista, Faculdade de Medicina da
de 2005
Universidade Federal do Ceará, Fortaleza,
Ceará
Pedro Palha, estudante da Licenciatura de 6/10/04 a Fevereiro Técnicas laboratoriais
Anatomia
Patológica,
Citológica
e
de 2005
Tanatológica, Instituto Politécnico de Saúde do
Norte, Paredes, Portugal
•
Situação Actual
Concluído
Concluído
Concluído
Concluído
Concluído
Concluído
Concluído
Em andamento
Em andamento
Visitas de Curta Duração
Professor Marcello Franco, Universidade Federal de São Paulo, SP, Brasil
Professor Francisco Valdeci Ferreira, Universidade Federal do Ceará, CE, Brasil
Prof. Adhemar Longatto, Instituto Adolfo Lutz, São Paulo, SP, Brasil
4. Publicações com material da U.P.S.
Soares R, Schmitt FC. Angiogenesis in brain tumors. Revista Neurologia 38: 3-8, 2004.
Gama A, Paredes J, Albergaria A, Gartner F, Schmitt F. P-cadherin expression in canine mammary tissues.
Journal of Comparative Pathology 130: 13-20, 2004.
Reis-Filho JS, Ricardo S, Gartner F, Schmitt FC. Bilateral gonadoblastomas in a dog with mixed gonadal
dysgenesis. Journal of Comparative Pathology 130: 229-233, 2004.
Mazeto GMFS, Oliveira MLCS, Padovani CR, Montenegro MR, Aragon FF, Schmitt FC. Thyroid cell
proliferation in Graves’disease. Use of MIB-1 monoclonal antibody. Acta Cytologica 48: 57-63, 2004.
Alves VAF, Bibbo M, Schmitt FC, Milanezi F, Longatto-Filho A. Comparison of manual and automated methods
of liquid-based cytology. Acta Cytologica 48: 187-193, 2004.
44
Medeiros R, Soares R, Vasconcelos A, Schmitt F, Lopes C. Glutathione S-transferase genotype GSTM1 as a
predictor of elevated angiogenic phenotype in patients with early onset breast cancer. Angiogenesis 7: 53-58,
2004.
Carvalho S, Silva AO, Milanezi F, Ricardo S, Leitão D, Amendoeira I, Schmitt FC. C-KIT and PDGFRA in
breast phyllodes tumours: overexpression without mutations ? Journal of Clinical Pathology 57: 1075-1079,
2004.
Costa C, Soares R, Schmitt FC. Angiogenesis: now and then. Acta Pathologica, Microbiológica et
Immunologica Scandinavica 112: 400-412, 2004.
5.
Exames realizados na U.P.S.
Nº total de exames: 12.613
Captura Híbrida: 29
Citologias: 8.379 (6.456 - Projecto de Santo Tirso)
Citologias Aspirativas: 1.633
Histológicos: 1.282
Histoquímicos: 93
Imuno-histoquímicos: 185
PCR por HPV: 5
Hibridização in situ: (Projecto ROCHE): 343
Genética Molecular e Citogenética
Diagnóstico de Sindrome de Prader-Willi: 80
Diagnóstico de Síndrome de Angelman: 38
Diagnóstico de Síndrome de X-frágil: 3
Diagnóstico de Doença de Crohn: 10
Estudo molecular de delecção 1p: 9
Estudo molecular de translocações: 10
Fenotipagem de -1 antitripsina: 146
Pesquisa de Amplificação de N-myc: 11
Pesquisa de Instabilidade de Microssatélites: 83
Pesquisa de Mutação no RET: 24
Pesquisa de Mutações do gene P53: 4
Pesquisa de Mutações nos genes BRCA1 e BRCA2: 12
Pesquisa de Mutações do gene OTC: 19
Pesquisa de Mutações do gene E-caderina: 7
Pesquisa de Mutações UBE3A: 14
Pesquisa de Mutações do gene SMAD4: 2
Citometria
Citometria de Fluxo: 25
Citometria de Imagem: 0
Casos em consulta*: 167
•
Dr.
Dr.
Dr.
Os casos em consulta foram oriundos das seguintes instituições:
A. Verhest - Institut Jules Bordet-Bruxelles-Bélgica
Abu Dhabi - Emirados Árabes Unidos
Agostinho Sanches - Hospital Senhora de Oliveira – Guimarães - Portugal
45
Dr.
Drª
Drª
Dr.
Dr.
Dr.ª
Dr.
Drª
Dr.
Dr.
Dr.
Dr.
Dr.ª
Dr.
Dr.
Dr.
Dr.ª
Dr.
Dr.
Dr.ª
Dr.
Drª
Dr.ª
Dr.
Dr.
Dr.
Dr.ª
Dr.
Dr.
Dr.
Drª
Dr.
Dr.
Albino Oliveira - Lab. Anatomia Patológica Dr. Albino Oliveira - Portugal
Ana Capdevila Puerta - A Corunã - Espanha
Ana Catarino - IPOFG-CROL, S.A. - Portugal
Aurel Perren - UniversitätsSpital Zürich - Suiça
Beatriz Homburg - CEDAP – Joinville – SC - Brasil
Beth Stenwig - University Hospital-The Norwegian Radium Hospital – Oslo - Noruega
Cardoso de Oliveira - Ordem do Carmo – Porto - Portugal
Carla Carrilho - Hospital Central de Maputo-Moçambique
Carlos Alberto Ribeiro - Belo Horizonte – MG - BRASIL
Carlos Alvarez Alvarez - Centro Médico Povisa – Coruña - Espanha
Carlos Prada Puentes - The Penine Acute Hospitalds NHS Trust - UK
Chantel Donne - França
Christine Dellau Vieira - Hospital Distrital Santarém, S.A. - Portugal
Christophe Duc - Institut Central dês Hôpitaux Valaisans - Suiça
Christophe Girardet - Institut Central des Hôpitaux Valaisans – Sion - Suiça
Consuelo Antunes B. Lins - Recife – PE - Brasil PE - Brasil
Daisy Lima - Centro Integrado de Colposcopia e Citopatologia – Recife – PE - Brasil
Eduardo Studart - Laboratório Silvany Studart - Hospital Português – Bahia - Brasil
Élbio C. de Paula - GOIÂNIA - GOIAS BRASIL
Eliana Chaves Salomão - Uberlândia – MG - Brasil
Emilio Marcelo Pereira - Laboratório Salomão e Zoppi – SP - Brasil
Fátima Magalhães - Unidade Local de Saúde de Matosinhos - Portugal
Francesa Felipo - Hospital Rojo Villanova - Espanha
Fred Ellinger - SP - Brasil
Gustavo Barbosa - SP - Brasil
H. Van Dijck - Laboratorium Voor Pathologische Ontleedkunde - Bélgica
Ierecê Aymoré - CLAP - Claudio Lemos Anat. Patológica – RJ - Brasil
Jannicke M. Berland - Stavanger - Noruega
João Carlos Coelho Filho - Fundação José Silveira – Bahia - Brasil
José Anselmo Coimbra Lopes - Lab. Jac. Lopes – Maranhão - Brasil
José Cameselle Teijeiro - Hospital Clínico Universitário - Santiago de Compostela - Espanha
José Dinis - Porto - Portugal
Júlio Defaveri - UNESP – Univ. Estadual Paulista “Júlio Mesquita” – SP - Brasil
Dr.ª
Dr.ª
Dr.
Dr.
Dr.
Dr.
Prof.
Dr.
Dr.ª
Drª
Dr.ª
Dr.ª
Krystyna Kotanska-Groholt - The Norwegian Radium Hospital - Norway
Linda Giannikaki - University Hospital of Crete - Crete - Grécia
Luciano Neder Serafini - H.C.F.M.R.P.-USP – Ribeirão Preto - Brasil
M. Akif Demir - Celal Bayar University - School of Medicine – Manisa – Turquia
Maecelo Alvarenga - Instituto de Patologia de Campinas, Ltda – SP - Brasil
Manuel Rodriguez Justo - University College London Hospitals – London - UK
Marcello Franco - São Paulo - Brasil
Drª
Dr.ª
Dr.ª
Drª
Drª
Dr.ª
Dr.ª
Dr.
Maria João Andrade - Hospital S. Teotónio – Viseu - Portugal
Maria José Brito - Hospital Garcia da Orta, SA., Almada - Portugal
Maria José Lorenzo Patiño - Complexo Hospitalario Juan Canalejo - Espanha
Maria Teresa Dias de Carvalho - Hospital Distrital de Viseu - Portugal
Marianne Heimann - Institut de Pathologie de Génétique - Bélgica
Marie Ludvikova - Medical Faculty Hospital – República Checa
Mary Toner - University Teaching Hospit. Trinity College Dublin - Irlanda
Müller Höcker - Pathologishes Institut der LMU – München - Alemanha
Margarida Teixeira - Hospital S. Marcos Braga - Portugal
Margarita González Cuesta - A Coruña - Espanha
Maria Cláudia Vaz Diniz - Campina Grande – Paraíba - Brasil
Maria Carmo Carvalho de Abreu e Lima - Recife - Brasil
46
Dr.
Dr.
Dr.ª
Dr.
Dr.ª
Dr.
Dr.
Dr.
Prof.
Dr.
Dr.
Dr.
Dr.
Dr.ª
Drª
Dr.
Dr.
Drª
Dr.ª
Dr.ª
Drª
Dr.
Dr.
Dr.
Dr.
Müller-Hermelink - Inst. Universität Würzburg - Alemanha
Nicolas de Saint Aubain - Institut Jules Bordet - Bélgica
Patricia Sabino Matos - FCM/ Unicamp – Campinas - Brasil
Paul Komminoth - Institut Für Pathologie – Baden - Suiça
Paula Guerra - S.M.A.S. - Portugal
Paulo Figueiredo - I.P.O. – Coimbra - Portugal
Paulo Roberto Grimaldi Oliveira - Paraiso – SP - Brasil
Pedro Oliveira - I.P.O. – Lisboa - Portugal
R. Heimann - Brussels - Bélgica
Robert Lemoine - INAP – Neuchatel - Suiça
Rogério de Almeida Ribeiro - Brasília - Brasil
Ronald de Krijger - Erasmus MC - Holanda
Rosângela Deliza - Campinhas - Brasil
Salete Silva - Hospital Fernando Fonseca - Portugal
Sérgio Tavolaro Pereira - Santos – SP - Brasil
Serpil Dizbay Sak - Ankara University Faculty of Medicine - Sihhiye-Ankara-Turquia
Sofia Loureiro dos Santos - Hospital Garcia da Orta - Portugal
Sónia Lima Marcena - Centro de Saúde Dr Jose Augusto Barreto - Aracaju - Brasil
Susanne B-Wildhagen - Stavanger - Noruega
Teresa Ribas Ariño - Hospital de León - Espanha
Tissier-Rible - Paris - França
Tom Clarke - Royal Devon and Exeter Healthcare - UK
Valdeci Ferreira - Fortaleza – Ceará - Brasil
Violante Assis - Faculdade de Medicina da Universidade Federal – MG - Brasil
6.
Controle de Qualidade
•
•
Data de Instituição
19-01-1998
Membros
• Prof. Fernando Schmitt
• Drª Fernanda Milanezi
• D. Susana Silva
•
•
Nº de Casos Revistos em 2004 – 302 casos de patologia cirúrgica e citopatologia (não ginecológica)
Nº de Casos Revistos em 2004 de citologias ginecológicas: 1404
•
Principais Conclusões no Final do 7º Ano:
Em relação aos anos anteriores houve uma modificação completa do Sistema de Controle de Qualidade com a
implementação do sistema informático Sislab a partir de Abril de 2004. Este programa permite um controle
significativamente melhor das actividades de rotina e fornece dados detalhados de vários aspectos da
produção dos exames. Uma das primeiras consequências foi o da criação de um sistema de controle de
qualidade individualizado para a citologia ginecológica de acordo com o requerido pela CAP, razão pela qual
passaremos a partir deste ano a fornecer os dados em separado.
Na análise do controle de qualidade de patologia cirúrgica e citopatologia não ginecológica (em 2004, de
Janeiro a Março inclui citologias ginecológicas, já que o Sislab passou a funcionar a partir de Abril), os
principais resultados foram os seguintes:
1.
2.
3.
A concordância diagnostica total entre o diagnóstico original e o revisto pela Comissão de Controlo de
Qualidade manteve-se nos mesmos níveis alcançados em 2003, com ligeira melhoria (99,6%).
A partir deste ano passamos a analisar o 1º circuito dos exames (macroscopia e colheita de biopsia
aspirativa). Encontramos discordâncias em oito dos 302 casos analisados (3,5%).
Em relação a qualidade do corte histológico, 96,2% dos casos foram considerados bons ou satisfatórios.
47
4.
5.
6.
Em relação a qualidade da coloração, 96,6% dos casos foram considerados excelentes, bons ou
satisfatórios.
O número de exames não codificados caiu para zero (2,2% em 2003), uma das metas atingidas com a
implementação do novo sistema informático e do sistema de codificação SNOMED. Houve erro de
codificação em 0,9% dos casos.
O cálculo do “turn-around-time” passou a ser controlado pelo sistema informático desde o registo do
material na UPS até sua expedição, não sendo possível comparar com os dados anteriores que diziam
respeito a realização dos exames e elaboração dos relatórios. Nos exames mais frequentes estes tempos
foram de 2,8 dias para biopsias aspirativas com colheita, 4,9 dias para histológicos de biopsias, 9,5 dias
para histológicos de peças, 7,2 dias para biopsias hepáticas com imunohistoquímica. Elaboramos uma
meta para redução significativa nos tempos relativos aos exames histológicos em 2005.
Na análise do controle de qualidade de citologia ginecológica (Abril a Dezembro de 2004), os principais
resultados foram os seguintes:
7.
De acordo com os critérios estabelecidos no Procedimento Regulamentador P.R.MED 01-02 Análise
microscópica de citologia ginecológica, dentre os casos observados pelo citotécnico foram revistos 1404
casos, sendo o diagnóstico concordante em 93,3%. Dos 6,7% dos casos discordantes, em 81,9% dos
casos o citopatologista reduziu a gravidade da alteração assinalada pelo citotécnico, enquanto em 18,1%
o diagnóstico final foi mais grave do que o previamente assinalado pelo citotécnico. Tais números nos
próximos anos poderão dar ideia do tipo de formação a que deve ser submetido o citotécnico.
8. Em relação a qualidade da coloração, 17,2% dos casos foram considerados excelentes/bons, 76,3%
satisfatórios, 6,5% regulares e 4% não satisfatórios.
9. O número total de citologias consideradas não satisfatórias para a análise foi de 4,1% (2,7% em 2003), o
que denota cada vez mais a necessidade da implementação da citologia em meio-líquido no rastreio de
Santo Tirso, conforme chamamos a atenção desde 2000.
10. Ascus foi diagnosticado em 2,5% com um relação Ascus/Lesão de 3,2. Houve uma melhora significativa
deste parâmetro em relação a 2003 (6,4% e 7,9) e o atingimento dos valores recomendados pela CAP (Nº
de Ascus inferior a 4,5% e relação Ascus/Lesão de 2,0).
11. De acordo com o referido no ano passado continuamos no projecto para Acreditação da UPS pelo Colégio
Americano de Patologistas. Dentro do âmbito deste processo em 2003 iniciamos nossa participação nos
programas de controlo de qualidade externo da CAP, relativos a patologia cirúrgica, imuno-histoquímica,
citologia ginecológica e citologia não-ginecológica. Também participamos no controlo de qualidade
externo das técnicas de imuno-histoquímica do UK-NEQAS e do Programa de Qualidade da Sociedade
Brasileira de Patologia (PIQ). Todas estas actividades estão registadas de acordo com o Procedimento
Regulamentador P.R.MED.05-01- Programas Externos de Educação e Avaliação Contínua.
48
5b) UNIDADE DE PRESTAÇÃO DE SERVIÇOS (UPSi)
Nº de exames requeridos:
Pedidos cancelados:
Em curso (ainda por efectuar ou por falta de 1 ou mais colheitas):
175
0
19
Tipos de exame:
Caracterizações/Identificações genéticas:
2
Paternidades
Com 1 pretenso pai, sem análise da mãe:
Com 2 filhos:
Com 1 pretenso pai e com análise da mãe:
Com 2 filhos:
Com 2 pretensos pais e com análise da mãe:
26
6
126
4
4
Outros parentescos:
Pareceres
5
2
Locais de requisição:
Local
Porto
Vila Nova de Gaia
Lisboa
Sta. Mª Feira
Santo Tirso
Penafiel
Bragança
Aveiro
Macedo de Cavaleiros
Vila do Conde
Ponta Delgada
Ponte da Barca
Arouca
Baião
Póvoa de Varzim
Ribeira Brava (Madeira)
Braga
Esposende
Paredes de Coura
Total
Tribunais
50
23
0
9
3
6
3
0
3
5
0
0
1
0
2
0
0
1
0
106 (61%)
Particulares
16
0
42
0
0
0
0
4
0
0
1
1
0
1
0
1
2
0
1
69 (39%)
Total
66
23
42
9
3
6
3
4
3
5
1
1
1
1
2
1
2
1
1
175
Tipos de requerentes
Tribunais
Averiguações Oficiosas de Paternidade
Outras
Particulares
103
3
69
Nº exames por requerente
Clientes
Total
49
Tribunais:
Trib. Família e Menores do Porto
Trib. Família e Menores de Vila Nova de Gaia
Trib. Santa Maria da Feira
Trib. Santo Tirso
Trib. Penafiel
Trib. Bragança
Trib. Macedo de Cavaleiros
Trib. Vila do Conde
Trib. Arouca
Trib. Póvoa de Varzim
Trib. Esposende
50
23
9
3
6
3
3
5
1
2
1
Particulares
Clínica Dr. Joaquim Chaves - Lisboa
GDPN - Porto
Dr. Alberto Ferreira Neves - Aveiro
Outros
41
15
4
9
Total
175
50
5c) UPSs – Unidade de Prestação de Serviços de Susceptibilidade Genética
Director: José Carlos Machado
A UPSS foi criada com dois objectivos fundamentais:
1- Dotar o IPATIMUP de capacidade acrescida, na qualidade de prestador de serviços, na área dos
testes de diagnóstico genético e susceptibilidade genética;
2- Promover a transferência de tecnologia e “know-how” resultantes da actividade científica do
IPATIMUP para o estabelecimento de novos testes de diagnóstico genético e susceptibilidade
genética.
A UPSS foi criada em 2004 e a sua actividade imediata visou criar condições mínimas para o arranque da
unidade. A lista seguinte é um sumário daquilo que foi alcançado até agora:
•
A UPSS iniciou a sua actividade como prestador de serviços oferecendo os seguintes testes:
o
Genotipagem de Helicobacter pylori e polimorfismos pró-inflamatórios associados com risco
de carcinoma gástrico;
o
•
Detecção e tipagem do vírus do papiloma humano.
A UPSS obteve o seu espaço físico próprio no IPATIMUP. Este foi um passo crucial que
providenciará à UPSS uma maior autonomia, melhorará a sua capacidade de gestão e reduzirá o
potencial de conflito com a actividade de investigação a decorrer no IPATIMUP.
•
A UPSS iniciou o processo de incorporação de testes de diagnóstico genético e susceptibilidade
genética já existentes no IPATIMUP. Esperamos que esta iniciativa permita a qualquer Investigador
do IPATIMUP interessado em realizar este tipo de testes por rotina, o acesso a um espaço,
equipamento e estrutura organizativa comuns.
•
A UPSS, em representação do IPATIMUP, estabeleceu um acordo com a empresa “GENETEST –
Prestação de serviços de testes de diagnóstico genético, S.A.”. Esta empresa foi co-fundada por
vários investigadores do IPATIMUP e prevemos que constitua um parceiro fundamental para
melhorar a nossa capacidade de comercializar serviços e realizar actividade de investigação e
desenvolvimento em consórcio.
•
A UPSS, em representação do IPATIMUP e em consórcio com a GENETEST, submeteu um projecto
de investigação à Agência de Inovação intitulado “Teste de susceptibilidade genética para
determinação de risco de desenvolvimento de Doença de Crohn na população Portuguesa”. Este
projecto foi aprovado e encontra-se em fase final de homologação.
Em 2005 o nosso principal objectivo será consolidar a actividade da UPSS. As tarefas específicas a que nos
propomos incluem:
•
A organização de um “reception front desk”;
51
•
O estabelecimento de uma base de dados para gestão de testes;
•
A contratação de um técnico de laboratório;
•
O desenvolvimento da parceria com a GENETEST.
52
6. DOUTORAMENTOS
53
Teses de doutoramento
Mafalda Pinto, Microbiologista, doutorou-se em Biologia Humana na Faculdade de Medicina da Universidade
do Porto. Biologia Humana/Oncobiologia. Janeiro 2004 (Orientador no IPATIMUP: Professora Raquel Seruca).
Título da Tese: “The mutator phenotype in sporadic gastric carcinomas: causes and consequences.”
Trabalhos
I. MSI-L gastric carcinomas have some of the molecular features of MSI-H carcinomas but not their
clinicopathological profile. Mafalda Pinto, Carla Oliveira, José Carlos Machado, Luís Cirnes, João
Tavares, Fátima Carneiro, Richard Hamelin, Robert Hofstra, Raquel Seruca, Manuel SobrinhoSimões. Laboratory Investigation 2000, 80: 1915-1923.
II. Promoter methylation of TGF-beta receptor I and mutation of TGF-beta receptor II are frequent events
in MSI sporadic gastric carcinomas. Mafalda Pinto, Carla Oliveira, Luís Cirnes, José Carlos Machado,
Maria Ramires, Ana Nogueira, Fátima Carneiro, Raquel Seruca. Journal of Pathology 2003, 200:3238.
III. Concurrent hypermethylation of gene promoters is associated with MSI-H phenotype and diploidy in
gastric carcinomas. Beatriz Carvalho, Mafalda Pinto, Luís Cirnes, Carla Oliveira, José Carlos
Machado, Gianpaolo Suriano, Richard Hamelin, Fátima Carneiro, Raquel Seruca. European Journal
of Cancer 2003, 39(9):1222-1227.
IV. MBD4 mutations are rare in gastric carcinomas with microsatellite instability. Mafalda Pinto, Ying Wu,
Rob Mensink, Luís Crines, Edwin Verlind, Gianpaolo Suriano, Robert Hofstra, Raquel Seruca. Cancer
Genetics and Cytogenetics 2003, 145: 103-107.
V. Frequent Ki-ras mutations in gastric tumours of the MSI phenotype. Caroline Brennetot, Mafalda
Pinto, Carla Oliveira, Simó Schwartz Jr, Raquel Seruca, Alex Duval and Richard Hamelin.
Gastroenterology 2003, 125: 1282-1283.
VI. BRAF mutations characterize colon but not gastric cancer with mismatch repair deficiency. Carla
Oliveira, Mafalda Pinto, Alex Duval, Caroline Brennetot, Enric Domingo, Eloi Espin, Manel Armengol,
Hiroyuki Yamamoto, Richard Hamelin, Raquel Seruca and Simó Schwartz Jr. Oncogene 2003, 22:
9192-9196.
Raquel Soares, bioquímica, mestre em Oncobiologia, doutorou-se em Biologia Humana na Faculdade de
Medicina da Universidade do Porto em Janeiro de 2004 (Orientador no IPATIMUP: Prof. Fernando Schmitt)
Título da Tese: Mechanisms of Hormone Regulation in Angiogenesis
Trabalhos:
I. Marinho A, Soares R, Ferro J, Lacerda M, Schmitt FC. Angiogenesis in breast cancer is related to age
but not to other prognostic parameters. Pathol Res Pract, 1997; 193: 267- 273.
II. Schmitt FC, Soares R. TGF and angiogenesis. Am J Surg Pathol, 1999; 23: 358-359
III. Schmitt FC, Soares R. Hormonal control of angiogenesis in breast cancer: TGF&#61537, a missed link?
The Breast, 1999; 8: 154.
IV. Soares R, Botelho M, Silva C, Ferro J, Wagner R, Schmitt FC. Expression of TGF and EGFR
in breast cancer and its relation to angiogenesis. The Breast J, 2000; 6: 171- 177.
V. Costa C, Soares R, Reis-Filho J, Leitão D, Amendoeira I, Schmitt FC. Cyclo-oxygenase-2 expression is
associated with angiogenesis and lymph node metastasis in human breast cancer. J Clin Pathol, 2002;
55: 429- 434.
VI. Soares R, Reis-Filho J, Gärtner F, Schmitt FC. VEGF, TGF and estrogen receptors: possible
cross-talks and interactions. Am J Pathol, 2002; 160: 381- 383.
VII. Soares R, Guo S, Russo J, Schmitt FC. Role of the estrogen antagonist ICI182,780 in vessel assembly
and apoptosis of endothelial cells. Ultrastruct Pathol, 2003; 27: 33- 39.
VIII. Soares R, Guo S, Gärtner F, Schmitt F, Russo J. 17-Estradiol-mediated vessel assembly and
stabilization in tumor angiogenesis requires TGFbeta and EGFR crosstalk. (in press in Angiogenesis)
IX. Soares R, Balogh G, Guo S, Gärtner F, Russo J, Schmitt F. Evidence for the Notch signaling pathway
on the role of estrogen in angiogenesis (Submitted paper).
54
Ana Preto, Farmacêutica, aluna do Programa GABBA, doutorou-se em Biologia Humana pela Faculdade de
Medicina da Universidade do Porto em Janeiro de 2004 (Orientador no IPATIMUP: Professor Manuel
Sobrinho-Simões).
Título da Tese: “Identification of potential mechanisms of tolerance to wild-type p53 in human carcinogenesis
using thyroid neoplasia as a prototype”.
Trabalhos :
I. Preto A, Reis Filho JS, Ricardo S, Soares P. “P63 expression in papillary and anaplastic carcinomas of
the thyroid gland: lack of an oncogenic role in tumorigenesis and progression”. Pathology Research and
Practice, 198: 449-454 (2002).
II. Reis Filho JS, Preto A, Soares P, Ricardo S, Cameselle-Teijeiro J, Sobrinho-Simões M. “p63 in Solid
cell nests: further evidence for a stem cell origin”. Modern Pathology, 16: 43-48 (2003).
III. Preto A, Singhrao S, Haughton M, Kipling D, Wynford-Thomas D and Jones CJ. “Telomere erosion
triggers growth arrest but not cell death in human cancer cells retaining wild-type p53: implication for
anti-telomerase therapy”. (Oncogene, in press).
IV. Preto A, Cameselle-Teijeiro J, Moldes-Boullosa J, Soares P, Cameselle-Teijeiro JF, Silva P, Reis-Filho
JS, Reyes-Santías RM, Alfonsín-Barreiro N, Forteza J, Sobrinho-Simões M. “Telomerase expression
and proliferative activity suggest a stem cell role for thyroid solid cell nests”. ( Modern Pathology, in
press).
Luísa Azevedo, bióloga, doutorou-se em Biologia na Faculdade de Ciências da Universidade do Porto em
Fevereiro de 2004 (Orientador no IPATIMUP: Prof. António Amorim)
Título da Tese: "A deficiência em ornitina transcarbamilase: diagnóstico, espectro mutacional e análise
haplotípica”.
Trabalhos
1. Azevedo, L., Climent, C., Vilarinho, L., Amorim, A. (2004). Evidence for mutational cis-acting factors affecting
mutagenesis in the ornithine transcarbamylase gene. Hum Mutat 24:273.
2. Azevedo, L, Stolnaja, L, Tietzeova, E, Hrebicek, M., Hruba, E., Vilarinho, L., Amorim, A., Dvorakova, L.
(2003). New polymorphic sites within ornithine transcarbamylase (OTC) gene: population genetics studies and
implications for diagnosis. Mol Genet Metab 78: 152-157.
3. Azevedo, L., Calafell, F., Vilarinho, L., Amorim, A. (2002). Haplotype analysis and phylogeny of ornithine
transcarbamylase polymorphisms. Ann. Hum. Genet. 66: 379-385.
4. Azevedo, L., Vilarinho, L., Teles, E.L., Amorim, A. (2002). Ornithine transcarbamylase deficiency: a novel
splice site mutation in a family with meiotic recombination and a new useful SNP for diagnosis. Mol Genet
Metab 76: 68-70.
5. Azevedo, L., Vilarinho, L., Teles, E.L., Martins, E., Cenni, B., Wermuth, B., Amorim, A. Mutational spectrum
and linkage disequilibrium in the ornithine transcarbamylase gene (in press at Ann Hum Genet).
6. Azevedo, L., Suriano, G., Amorim, A. Ornithine transcarbamylase sequence and function comparisons in
hominoids (submitted to Mol Biol Evol).
Joana Paredes, bióloga, doutorou-se em Biologia Humana na Faculdade de Medicina da Universidade do
Porto em Outubro de 2004 (Orientador no IPATIMUP: Prof. Fernando Schmitt)
Título da Tese: "P-caderina – sua Importância na Carcinogénese e Progressão do Cancro da Mama"
Trabalhos:
55
1. Paredes J, Milanezi F, Viegas L, Amendoeira I, and Schmitt F. 2002. P-cadherin expression is associated
with high-grade ductal carcinoma in situ of the breast. Virchows Arch 440:16-21.
2. Reis-Filho JS, Cancela Paredes J, Milanezi F, and Schmitt FC. 2002. Clinicopathologic implications of Ecadherin reactivity in patients with lobular carcinoma in situ of the breast. Cancer 94:2114-5.
3. Paredes J, Milanezi MF, Reis-Filho JS, Leitão D, Athanazio D, and Schmitt FC. 2002. Correlation between Pcadherin and estrogen receptor expression in breast cancer. J Bras Patol Med Lab 38:307-313.
4. Gama A, Paredes J, Milanezi MF, Reis-Filho JS, Gartner F, and Schmitt FC. 2002. P-cadherin expression in
canine lactating mammary gland. J Cell Biochem 86:420-1.
5. Paredes J, Milanezi F, Reis-Filho JS, Leitão D, Athanazio D, and Schmitt F. 2002. Aberrant P-cadherin
expression: is it associated with estrogen-independent growth in breast cancer? Pathol Res Pract 198:795-801.
6. Reis-Filho JS, Milanezi F, Paredes J, Silva P, Pereira EM, Maeda SA, de Carvalho LV, and Schmitt FC.
2003. Novel and classic myoepithelial/stem cell markers in metaplastic carcinomas of the breast. Appl
Immunohistochem Mol Morphol 11:1-8.
7. Gama A, Paredes J, Albergaria A, Gartner F, and Schmitt F. 2004. P-cadherin expression in canine
mammary tissues. J Comp Pathol 130:13-20.
8. Paredes J, Stove C, Stove V, Milanezi F, Van Marck V, Derycke L, Mareel M, Bracke M, and Schmitt F.
2004. P-Cadherin Is Up-Regulated by the Antiestrogen ICI 182,780 and Promotes Invasion of Human Breast
Cancer Cells. Cancer Res 64: 8309–8317.
9. Van Marck V, Stove C, Stove V, Paredes J, and Bracke M. 2004. P-cadherin promotes cell-cell adhesion and
counteracts invasion in human melanoma. Cancer Res (submitted).
10. Paredes J, Albergaria A, Oliveira J, Ricardo S, Leitão D, Serra V, Lacerda M, Cameselle-Teijeiro J,
Milanezi F, and Schmitt FC. 2005. Aberrant P-cadherin expression is associated with CDH3 promoter
methylation status and is an indicator of clinical outcome in invasive breast carcinomas. Clin Cancer Res
(submitted).
56
7. TRABALHOS PUBLICADOS OU ACEITES PARA PUBLICAÇÃO
57
7a. Artigos científicos
1.
Alves C, Gusmão L, Damasceno A, Soares B, Amorim A . Contribution for an African
autosomic STR database (AmpF/STR Identifiler and Powerplex 16 System) and a report on
genotypic variations, Forensic Sci Int 139: 201-205, 2004
1,6
2.
Alves S, Rocha J, Amorim A, Prata MJ . Tracing the origin of the most common thiopurine
methyltransferase (TPMT) variants: preliminary data from the patterns of haplotypic
association with two CA repeats. Ann Hum Genet 68:313-23, 2004
3,1
3.
Alves VAF, Bibbo M, Schmitt FC, Milanezi F, Longatto-Filho A. Comparison of manual and
automated methods of liquid-based cytology. Acta Cytol 48: 187-193, 2004.
1,0
4.
Araujo, R; Rodrigues, AG. Variability of germinative potential among pathogenic species of
Aspergillus. J Clin Microbiol , 42: 4335-4337, 2004
3,5
5.
Araujo, R; Rodrigues, AG; Pina-Vaz, C. A fast, practical and reproducible procedure for the
standardization of the cell density of an Aspergillus suspension J Med Microbiol 53: 783-786,
2004
2,0
6.
Azevedo L, Climent C, Vilarinho L, Calafell F, Amorim A. Evidence for mutational cis-acting
factors affectingmutagenesis in the ornithine transcarbamylase gene. Hum Mutat 24:273, 2004.
6,3
7.
Badhwar A, Berkovic SF, Dowling JP, Gonzalez M, Narayanan S, Brodtmann A, Berzen L,
Caviness J, Trekwalder C, Winkelmann J, Rivest J, Lambert M, Hernandez-Cossio O,
Carpenter S, Andermann F, Andermann E: Action myoclonus-renal failure syndrome:
characterization of a unique cerebro-renal disorder. Brain 127: 2173-2182, 2004.
8
8.
Beleza S, Alves C, Reis F, Amorim A, Carracedo A, Gusmão L . 17 STR data (AmpF/STR
Identifiler and Powerplex 16 System) from Cabinda (Angola). Forensic Sci Int 141: 193-196.
1,6
9.
Brooks-Wilson AR, Kaurah P, Suriano G, Leach S, Senz J, Grehan N, Butterfield YS, Jeyes J,
Schinas J, Bacani J, Kelsey M, Ferreira P, MacGillivray B, MacLeod P, Micek M, Ford J,
Foulkes W, Australie K, Greenberg C, LaPointe M, Gilpin C, Nikkel S, Gilchrist D, Hughes R,
Jackson CE, Monaghan KG, Oliveira MJ, Seruca R, Gallinger S, Caldas C, Huntsman D.
Germline E-cadherin mutations in hereditary diffuse gastric cancer: assessment of 42 new
families and review of genetic screening criteria. J Med Genet. 41:508-17, 2004.
6,4
10. Carneiro F, Huntsman D, Smyrk T, Owen DA, Seruca R, Pharoah P, Caldas C, Manuel
Sobrinho-Simões M: Model of early development of diffuse gastric cancer in E-cadherin
mutation carriers and its implications for patient screening. J Pathol 203: 681-687, 2004.
4,9
11. Carneiro F, Huntsman DG, Smyrk T, Owen D, Seruca R, Pharoah P, Caldas C, Sobrinho
Simões M: Pathology of inherited gastric cancer. Pathol Internat 54 (Suppl. 1): S285-S286,
2004.
1,2
12. Carneiro F, Oliveira C, Suriano G, Huntsman D, Caldas C, Seruca R: Hereditary diffuse gastric
cancer. Pathol Internat 54 (Suppl. 1): S193-S203, 2004.
1,2
13. Carneiro F: Qual o papel da biópsia hepática? J Port Gastroenter 11: 39-40, 2004.
-
14. Carrilho C, Alberto M, Buane L, David L: Keratins 8, 10, 13, and 17 are useful markers in the
3,7
58
diagnosis of human cervix carcinomas. Hum Pathol 35: 546-551, 2004.
15. Carvalho R, Milne AN, van Rees BP, Caspers E, Cirnes L, Figueiredo C, Offerhaus GJ,
Weterman MA. Early-onset gastric carcinomas display molecular characteristics distinct from
gastric carcinomas occurring at a later age. J Pathol 204:75-83, 2004.
4,9
16. Carvalho S, Silva AO, Milanezi F, Ricardo S, Leitão D, Amendoeira I, Schmitt FC. C-KIT and
PDGFRA in breast phyllodes tumours: overexpression without mutations? J Clin Pathol 57:
1075-1079, 2004.
3,0
17. Costa C, Soares R, Schmitt FC. Angiogenesis: now and then. APMIS 112: 400-412, 2004
0,9
18. Cruz J, Reis JS, Lopes JM. Malignant peripheral nerve sheath tumour-like primary cutaneous
malignant melanoma. J Clin Pathol 57: 218-220, 2004
3,0
19. De Souza Goes AC, De Carvalho EF, Gomes I, DA Silva DA, Gil EH, Amorim A, Gusmao L . 2,0
Population and mutation analysis of 17 Y-STR loci from Rio de Janeiro (Brazil). Int J Legal
Med. 2004 Nov 24; [Epub ahead of print]
20. Dias-Pereira P, Carvalheira J, Gartner F. Cell proliferation in feline normal, hyperplastic and
neoplastic mammary tissue – na immunohistochemical study. Vet J 168: 180-185, 2004.
1,3
21. Domingo E, Espín E, Armengol M, Oliveira C, Pinto M, Duval A, Brennetot C, Seruca R,
Hamelin R, Yamamoto H, Schwartz C Jr: Activated BRAF targets proximal colon tumours with
mismatch deficiency and hMLH1 inactivation. Genes Chromosome Cancer 39: 138-142, 2004.
4,2
22. Domingo E, Laiho P, Ollikainen M, Pinto M, Wang L, French AJ, Westra J, Frebourg T, Espin
E, Armengol M, Hamelin R, Yamamoto H, Hofstra RM, Seruca R, Lindblom A, Peltomaki P,
Thibodeau SN, Aaltonen LA, Schwartz S Jr. BRAF screening as a low-cost effective strategy
for simplifying HNPCC genetic testing. J Med Genet. 41:664-668, 2004
6,4
23. Faleiro-Rodrigues C, Lopes C. E-cadherin, CD44 and CD44v6 in squamous intraepithelial
lesions and invasive carcinomas of the uterine cervix: an immunohistochemical study.
Pathobiology 71:329-36, 2004
1,0
24. Fonseca AP, Extremina C, Fonseca AF, Sousa JC. Effect of subinhibitory concentration of
piperacillin/tazobactam on Pseudomonas aeruginosa. J Med Microbiol. 53: 903-10, 2004.
2,0
25. Gama A, Paredes J, Albergaria A, Gartner F, Schmitt F. P-cadherin expression in canine
mammary tissues. J Comp Pathol 130: 13-20, 2004.
1.3
26. García O, Martín P, Gusmão L, Albarrán C, Alonso S, De La Rua C, Flores C, Izagirre N,
Peñas R, Pérez JA, Uriarte I, Yurrebaso I, Alonso A . A Basque Country autochthonous
population study of 11 Y-chromosome STR loci. Forensic Sci Int. 145:65-68, 2004
1,6
27. Gaspar P, Seixas S, Rocha J. Genetic variation in a compound short tandem repeat/Alu
haplotype system at the SB19.3 locus: properties and interpretation. Hum Biol 76: 277-287,
2004
1,0
28. Keller G, Vogelsang H, Becker I, Plaschke S, Ott K, Suriano G, Mateus AR, Seruca R,
Biedermann K, Huntsman D, Doring C, Holinski-Feder E, Neutzling A, Siewert JR, Hofler H.
Germline mutations of the E-cadherin(CDH1) and TP53 genes, rather than of RUNX3 and
HPP1, contribute to genetic predisposition in German gastric cancer patients. J Med Genet.
41:e89, 2004.
6,4
59
29. Lakhani SR, Manek S, Penault-Llorca F, Flanagan A, Arnout L, Merrett S, McGuffog L, Steele
D, Devilee P, Klijn JG, Meijers-Heijboer H, Radice P, Pilotti S, Nevanlinna H, Butzow R, Sobol
H, Jacquemier J, Lyonet DS, Neuhausen SL, Weber B, Wagner T, Winqvist R, Bignon YJ,
Monti F, Schmitt F, Lenoir G, Seitz S, Hamman U, Pharoah P, Lane G, Ponder B, Bishop DT,
Easton DF. Pathology of ovarian cancers in BRCA1 and BRCA2 carriers. Clin Cancer Res 10:
2473-2481, 2004.
6,5
30. Lauwaet T, Oliveira MJ, Callewaert B, De Bruyne G, Mareel M, Leroy A.Proteinase inhibitors
TPCK and TLCK prevent Entamoeba histolytica induced disturbance of tight junctions and
microvilli in enteric cell layers in vitro. Int J Parasitol. 34:785-94, 2004
2,9
31. Lauwaet T, Oliveira MJ, De Bruyne G, Bruchhaus I, Duchene M, Mareel M, Leroy
A.Entamoeba histolytica trophozoites transfer lipophosphopeptidoglycans to enteric cell layers.
Int J Parasitol. 34: 549-56, 2004.
2,9
32. Lima J, Trovisco V, Soares P, Maximo V, Magalhaes J, Salvatore G, Santoro M, Bogdanova T,
Tronko M, Abrosimov A, Jeremiah S, Thomas G, Williams D, Sobrinho-Simoes M.. BRAF
mutations are not a major event in post-Chernobyl childhood thyroid carcinomas. J Clin
Endocrinol Metab 89:4267-4271, 2004.
5,9
33. Lima RT, Martins LM, Guimaraes JE, Sambade C, Vasconcelos MH: Specific downregulation
of bcl-2 and xIAP by RNAi enhances the effects of chemotherapeutic agents in MCF-7 human
breast cancer cells. Cancer Gene Ther 11:309-316, 2004.
3,7
34. Lloyd RV, Erickson LA, Casey MB, Lam KY, Lohse CM, Asa SL, Chan JKC, DeLellis RA,
Harach R, Kakudo K, LiVolsi VA, Rosai J, Sebo TJ, Sobrinho-Simoes M, Wenig BM, Lae ME.
Observer variation in the diagnosis of follicular variant of papillary thyroid carcinoma. Am J
Surg Pathol 28:1336-1340, 2004
4,5
35. Lopes AM, Calafell F, Amorim A . Microsatellite Variation and Evolutionary History of PCDHX/Y
Gene Pair Within the Xq21.3/Yp11.2 Hominid-Specific Homology Block. Mol Biol Evol. 21:
2092-2101, 2004
6,0
36. Lopez AM, Alvarez S, Gusmao L, Alves C, Mesa MS, Albentosa A, Arribas G, Lopez R, Barrio
PA, Amorim A, Arroyo-Pardo E . Population data for 16 Y-chromosome STRs in four
populations from Pyrenees (Spain). Forensic Sci Int 140:125-9, 2004
1,6
37. Magalhaes J, Sobrinho-Simoes M. Hyalinizing trabecular tumor of the thyroid displaying
neuroendocrine differentiation. Virchows Archiv 445: 98-98, 2004
2,4
38. Marcos NT, Pinho S, Grandela C, Cruz A, Samyn-Petit B, Harduin-Lepers A, Almeida R, Silva
F, Morais V, Costa J, Kihlberg J, Clausen H, Reis CA. Role of the human ST6GalNAc-I and
ST6GalNAc-II in the synthesis of the cancer- associated sialyl-Tn antigen. Cancer Res.
64:7050-7, 2004.
8,6
39. Martín P, García-Hirschfeld J, García O, Gusmão L, García P, Albarrán C, Sancho M, Alonso A
. A Spanish population study of 17 Y-chromosome STR loci. Forensic Sci Int. 139:231-235,
2004
1,6
40. Maximo V, Preto A, Crespo A, Soares P, Machado JC, Rocha AS, Sobrinho-Simoes M. Core I
gene is overexpressed in Hurthle and non-Hurthle cell microfollicular adenomas and follicular
carcinomas of the thyroid. BMC Cancer. 25; 4:12., 2004
1,7
41. Mazeto GMFS, Oliveira MLCS, Padovani CR, Montenegro MR, Aragon FF, Schmitt FC.
Thyroid cell proliferation in Graves’disease. Use of MIB-1 monoclonal antibody. Acta Cytol 48:
1,0
60
57-63, 2004.
42. Medeiros R, Soares R, Vasconcelos A, Schmitt F, Lopes C. Glutathione S-transferase
genotype GSTM1 as a predictor of elevated angiogenic phenotype in patients with early onset
breast cancer. Angiogenesis 7: 53-58, 2004.
-
43. Mesquita P, Almeida R, van Seuningen I, David L: Letter to the Editor -Coordinated expression 4,5
of MUC2 and CDX-2 in mucinous carcinomas of the lung can be explained by the role of CDX2 as transcriptional regulator of MUC2. Am J Surg Pathol 28: 1254-1255, 2004.
44. Nogueira AM, Marques T, Soares PC, David L, Reis CA, Serpa J, Queiroz DM, Rocha GA, Rocha AC: Lewis antigen expression in gastric mucosa of children: relationship with
Helicobacter pylori infection. Pediatr Gastroenterol Nutr 38: 85-91, 2004.
45. Oliveira C, de Bruin J, Nabais S, Ligtenberg M, Moutinho C, Nagengast FM, Seruca R, van
Krieken H, Carneiro F: Intragenic deletion of CDH1 as the inactivating mechanism of the wildtype allele in a HDGC tumour. Oncogene 23:2236-2240, 2004.
6,5
46. Oliveira C, Ferreira P, Nabais S, Campos L, Ferreira A, Cirnes L, Castro Alves C, Veiga I, 3,7
Fragoso M, Regateiro F, Moreira Dias L, Moreira H, Suriano G, Machado JC, Lopes C,
Castedo S, Carneiro F, Seruca R: E-Cadherin (CDH1) and TP53 rather than SMAD4 and
Caspase-10 germline mutations contribute to genetic predisposition in Portuguese gastric
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8b Capítulos de livros
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