Our research is focused in Barrett’s esophagus (BE), the premalignant condition for esophageal Adenocarcinoma (EA), which is characterized by the columnar metaplastic replacement of the normal squamous esophageal lining.
Our main area of research is the cellular differentiation of Barrett’s epithelium, its control and its relationship with malignant progression to EA. Presently, we are looking for the control of differentiation and proliferation by the characterization of the centrosome profile of BE and EA and looking for new prognostic biomarkers of BE.
1) Characterization of the differentiation profile of Barrett’s metaplastic epithelium and its relationship with cancer
Regarding BE cellular differentiation we have demonstrated that besides goblet cells, there are other metaplastic elements that exhibit intestinal differentiation and that these elements are also related to neoplasia. We have also demonstrated the presence of other phenotypes in BE, namely the gastric phenotype, and its association with malignant progression.
Concerning the biological meaning of the metaplastic segments with no intestinal metaplasia (IM) we have demonstrated that IM is a late event in the development of BE and that patients with no IM and with no recognized risk of progression may develop IM thus becoming risk patients. We are now interested in improving our understanding of the genetic control of the development and modulation of gastric and intestinal phenotypes in BE, namely, by the homeobox genes SOX2 and CDX2.
Considering the premalignant nature of BE metaplastic epithelium, we have demonstrated that BE is not only the precursor of most adenocarcinomas of the esophagus but also of most adenocarcinomas of the cardia.
2) The cause and consequence of centrosome abnormalities in Barrett’s tumorigenesis using paraffin-embedded specimens and cell lines
(in collaboration with Mónica Bettencourt-Dias – Cell Cycle Regulation Lab, IGC)
BE malignant progression involves major changes in cellular processes controlled by the centrosome, the primary microtubule-organizing centre in animal cells. Several studies have shown that cells from many cancers have abnormal centrosomes that are either correlated with tumour malignancy or considered an early event during tumorigenesis. However, a causative link between centrosome abnormalities and cancer remains elusive and its role on BE malignant transformation is still unknown.
In this study, we set out to investigate how centrosome defects contribute to tumorigenesis. We assessed the centrosome profile along the biopathogenic process of BE and associated neoplasia using paraffin-embedded dysplasia-free BE biopsies and esophagectomy specimens (from IPOLFG archives) as well as a panel of thirteen established cell lines derived from all steps of BE progression (two of the five adenocarcinoma-derived cell lines used were established in our lab and validated by the group of Winand Dinjens (Rotterdam) in a multicentre collaborative study).
Using double immunofluorescence labelling of centrosomes and centrioles we found that both numerical and size centriole defects progressively accumulate in BE tumorigenesis. Our findings suggest that centrosome abnormalities are related to Barrett’s progression and precede the onset of invasion. Correlation between these changes and the well-known genetic profile of the cell lines provides new clues into the permissive genetic background for such abnormalities (like p53 loss) and may impact on BE diagnosis, prognosis and treatment.
3) New prognostic markers of Barrett’s esophagus
(in collaboration with José Pereira Leal – Computational Genomics Lab, IGC)
The goal of this task was to perform a bioinformatics analysis of gene expression data sets in BE and EA and identify genes and pathways that defined malignant progression. The central goal of this task, is thus to identify clinically useful markers of cancer progression, and in particular BE and EA.
It was conducted an extensive survey of the literature and compiled a dataset of transcriptional alterations in the progression of BE to EA. Because of the nature of the data, standard pipelines of analysis were not suitable, which led us to develop a new bioinformatics framework for meta-analysis of gene expression data. With this, it was possible to identify a set of 19 genes that under the most conservative criteria can distinguish a BE that is likely to progress from a BE that will not progress to EA. These markers were validated by real time RT-qPCR on RNA from time-series cohort of IPOLFG patients that progressed from BE to EA. RT-qPCR on this group of patients identified CYR61 and WWTR1 over-expression years before the development of adenocarcinoma as compared to patients who have never faced malignant progression. These two markers were also validated by Immunohistochemistry.
The results suggest that alterations with prognostic value can be detected very early. This study allowed the identification of WWTR1 and CYR61 as having a potential role in early risk stratification of Barrett’s esophagus neoplastic progression and thus with prospects to impact patient management.
ONGOING RESEARCH PROJECTS
“CAUSES AND CONSEQUENCES OF CENTROSOME AND PLOIDY ABNORMALITIES IN HUMAN CANCER USING BARRETT'S ESOPHAGUS AS A MODEL”
Cancer and Centrosome Consortium:
Mónica Bettencourt-Dias, Instituto Gulbenkian de Ciência (Group Leader)
David Pellman, Harvard Medical School (PI)
Massimo Loda, Harvard Medical School (PI)
Paula Chaves, Instituto Português de Oncologia de Lisboa Francisco Gentil (PI)
José Pereira-Leal, Instituto Gulbenkian de Ciência (PI)
Funding: Harvard Medical School Portugal Program, 2009-2013
“ESÓFAGO DE BARRETT – CONTRIBUTOS PARA A DEFINIÇÃO DO CONCEITO E PARA O CONHECIMENTO DO RISCO DE PROGRESSÃO NEOPLÁSICA NUMA POPULAÇÃO PORTUGUESA”
Projecto de tese de doutoramento apresentado à FCS UBI
Dias Pereira, Instituto Português de Oncologia de Lisboa Francisco Gentil (PI).
Paula Chaves, Instituto Português de Oncologia de Lisboa Francisco Gentil.
PRIZES AND AWARDS
PRÉMIO PARA A 2ª MELHOR COMUNICAÇÃO oral na área da Ciência Básica, XXV Congresso Nacional de Gastrenterologia e Endoscopia Digestiva – 2005: FENÓTIPO CENTROSSÓMICO DO ADENOCARCINOMA DO ESÓFAGO E DA METAPLASIA DE BARRETT ADJACENTE
C Laranjeira, P Chaves, S Dias, AD Pereira, F Negreiro, AG Oliveira, CN Leitão, J Soares.
PRÉMIO PARA A 2ª MELHOR COMUNICAÇÃO oral na área clínica, Congresso Nacional de Gastrenterologia e Endoscopia Digestiva – 2005: ESÓFAGO REVESTIDO POR EPITÉLIO COLUNAR SEM METAPLASIA INTESTINAL: CARACTERÍSTICAS E SIGNIFICADO.
AD Pereira, P Chaves, A Suspiro, M Crespo, J Soares, CN Leitão.
PRÉMIO PARA A MELHOR COMUNICAÇÃO oral do IX Congresso Nacional de Anatomia Patológica – 2004: CARACTERIZAÇÃO MORFOLÓGICA DA JUNÇÃO ESCAMOSO- COLUNAR (LINHA Z) NO FETO.
RA Cabrera, L Ayala, C Rodrigues, M Crespo, P Borralho, AD Pereira, P Chaves, J Soares.
PRÉMIO PARA O MELHOR POSTER do XXI Congresso Nacional de Gastrenterologia e Endoscopia Digestiva – 2001: Alterações citogenéticas e da imunoexpressão da proteína p53: eventos precoces no esófago de Barrett
C Ribeiro, AD Pereira, P Chaves, A Gírio, A Suspiro, L Roque, CN Leitão, J Soares.
MELHOR TRABALHO DE CIÊNCIA BÁSICA, OESO – 6th World Congresss, 2000:
The enterocytic differentiation of Barrett’s epithelium with and without associated cancer.
P Chaves, C Cruz, P Cardoso, A. Suspiro, AD Pereira, JC Mendes Almeida, CN Leitão, J Soares.
PRÉMIO NACIONAL DE GASTRENTEROLOGIA, 1999: Esófago de Barrett: Resultados de um programa prospectivo de vigilância.
AD Pereira, A. Suspiro, P. Chaves, J C Mendes Almeida, CN Leitão.
PRÉMIO PARA O MELHOR VÍDEO do XV Congresso Nacional de Gastrenterologia e Endoscopia Digestiva – 1995: ESÓFAGO DE BARRETT – O DIAGNÓSTICO ENDOSCÓPICO.
AD Pereira, P. Chaves, A. Suspiro, FC Mira.
- Rosa I, Fidalgo P, Filipe B, Albuquerque C, Fonseca R, Chaves P, Pereira AD. Sporadic colorectal cancer: Studying ways to an end. United European Gastroenterol J. 2016 Apr;4(2):288-96. doi: 10.1177/2050640615599329. Epub 2015 Aug 6.PMID:27087959
- Dias Pereira A, Chaves P. (2012) COLUMNAR-LINED OESOPHAGUS WITHOUT INTESTINAL METAPLASIA: RESULTS FROM A COHORT WITH A MEAN FOLLOW-UP OF 7 YEARS. Aliment Pharmacol Ther 36(3):282-9.
- Chaves P, Dias Pereira A. (2010) THE BIOLOGICAL MEANING OF INTESTINAL METAPLASIA OF THE GASTROESOPHAGEAL JUNCTION. Int J Surg Pathol. 18:43-47.
- Boonstra JJ, van Marion R, Beer DG, Lin L, Chaves P, Ribeiro C, Pereira AD, Roque L, Darnton SJ, Altorki NK, Schrump DS, Klimstra DS, Tang LH, Eshleman JR, Alvarez H, Shimada Y, van Dekken H, Tilanus HW, Dinjens WN.(2010) VERIFICATION AND UNMASKING OF WIDELY USED HUMAN ESOPHAGEAL ADENOCARCINOMA CELL LINES. J Natl Cancer Inst. 102(4):271-4.
- Caygill CP, Zaninotto G, Rugge M, Bergman JJ, Caygill CP, Chaves P, Pereira AD, Dolina J, Fuchs KH, Gatenby P, Lerut A, Murray L, Regula J, Rugge M, Watson A, Wittmann T, Zaninotto G, Rizzetto C, Portale G.(2008) BARRETT’S REGISTRIES IN EUROPE: REPORT OF AN INTERNATIONAL WORKSHOP. Eur J Cancer Prev. 17(5):426-9. 2008.
- Pereira AD, Suspiro A, Chaves P.(2007) CANCER RISK IN BARRETT’S ESOPHAGUS. Eur J Gastroenterol Hepatol. 19(11): 915-918.
- Chaves P, Crespo M, Ribeiro C, Laranjeira C, Pereira AD, Suspiro A, Cardoso P, Leitão CN, Soares J. (2007) CHROMOSOMAL ANALYSIS OF BARRETT'S CELLS: DEMONSTRATION OF INSTABILITY AND DETECTION OF THE METAPLASTIC LINEAGE INVOLVED. Mod Pathol 20:788 – 796.
- Chaves P, Cruz C, Pereira AD, Suspiro A, Mendes Almeida JN, Leitão CN, Soares J. (2005) GASTRIC AND INTESTINAL DIFFERENTIATION IN BARRETT’S METAPLASIA AND ASSOCIATED ADENOCARCINOMA. Dis Esophagus 18:383 – 387.
- Chaves P, Cruz C, Cardoso P, Suspiro A, Pereira AD, Mendes Almeida JC, Leitão CN, Soares J. (2003) ENTEROCYTIC COLUMNAR NON–GOBLET CELLS OF BARRETT’S ESOPHAGUS - AN IMMUNOHISTOCHEMICAL DEMONSTRATION OF ASSOCIATION WITH MALIGNANT EVOLUTION. J Exp Clin Cancer Res 22:273 – 278.
- Suspiro A, Pereira AD, Afonso A, Chaves P, Soares J, Leitão CN. (2003) LOSS OF HETEROZYGOSITY ON CHROMOSES 17P AND 9P ARE FREQUENT EVENTS IN BARRETT’S METAPLASIA NOT ASSOCIATED WITH DYSPLASIA OR ADENOCARCINOMA. Am J Gastroenterol 98:728 – 734.
- Chaves P, Pereira AD, Cruz C, Suspiro A, Mendes Almeida JC, Leitão CN, Soares J. (2002) RECCURRENT COLUMNAR-LINED ESOPHAGEAL SEGMENTS - STUDY OF THE PHENOTYPIC CHARACTERISTICS USING INTESTINAL MARKERS. Dis Esophagus 15:282 – 6.
- Cabrera RA, Chaves P, Crespo M, Pereira AD, Mendes Almeida JC, Leitão CN, Soares J.(2002) ADENOCARCINOMA OF THE ESOPHAGOGASTRIC JUNCTION: COULD THE CHARACTERISTICS OF ADJACENT INTESTINAL METAPLASIA HELP ON THE UNDERSTANDING OF BIOPATHOGENESIS? Dis Esophagus 15:287 – 9.
- Pereira AD, Chaves P, Suspiro A, Mendes Almeida JC, Soares J, Leitão CN. (1999) ESÓFAGO DE BARRETT: RESULTADO DE UM PROGRAMA PROSPECTIVO DE VIGILÂNCIA. GE – J Port Gastrenterol 6:164- 172.
- Chaves P, Cardoso P, Mendes Almeida JC, Pereira AD, Leitão CN, Soares J. (1999) NON-GOBLET CELL POPULATION OF BARRETT’S ESOPHAGUS: AN IMMUNOHISTOCHEMICAL DEMONSTRATION OF INTESTINAL DIFFERENTIATION. Human Pathol 30:1291 – 1295.
- Pereira AD, Suspiro A, Chaves P, Saraiva A, Glória L, Mendes Almeida JC, Leitão CN, Mira FC. (1998) SHORT SEGMENTS OF BARRETT’S EPITHELIUM AND INTESTINAL METAPLASIA IN NORMAL APPEARING OESOPHAGOGASTRIC JUNCTIONS: THE SAME OR TWO DIFFERENT ENTITIES? Gut 42:659 – 662.
- Mendes Almeida JC, Chaves P, Pereira AD, Altorki NK. (1997) IS BARRETT'S ESOPHAGUS THE PRECURSOR OF MOST ADENOCARCINOMAS OF THE ESOPHAGUS AND CARDIA? A BIOCHEMICAL STUDY. Ann Surg 226:725 – 733, 1997.
Mónica Bettencourt-Dias, PhD
Cell Cycle Regulation Laboratory
Instituto Gulbenkian de Ciência, Lisboa, Portugal.
José Leal, PhD
Computational Genomics Laboratory
Instituto Gulbenkian de Ciência, Lisboa, Portugal.
David Pellman, MD, PhD
Dana-Farber Cancer Institute, Harvard Medical School, Boston, USA.
Leonor David, MD, PhD
Differentiation and Cancer
IPATIMUP, Porto, Portugal