Association of family risk and lifestyle/comorbidities in ovarian cancer patients

Summary Objectives: to analyze factors that might indicate familial predisposition for ovarian cancer in patients diagnosed with this disease. Methods: in a prospective single center cohort study at the Institute of Cancer of the State of São Paulo (ICESP), 51 women diagnosed with ovarian cancer were included. Familial predisposition for ovarian cancer was defined as having a higher than 10% chance of having a BRCA1/2 mutation according to the Manchester scoring system, a validated method to assess the likelihood of mutation detection. Each patient was interviewed with a standardized questionnaire on established risk factors for ovarian cancer and other factors that might influence the risk to develop ovarian cancer. Logistic regression analyses were performed to estimate the impact of the evaluated factors on the likelihood of mutation detection, by calculating odds ratios and 95% confidence intervals. Results: seventeen out of 51 patients had a family history of breast and/or ovarian cancer, four patients had a history of breast or endometrial cancer, 11 were diagnosed before the age of 50, and 12 presented a risk of familial predisposition to ovarian cancer higher than 10%. Patients with comorbidities, such as hypertension, diabetes, hormonal disorders, dyslipidemia and psychiatric conditions, presented a lower chance of having a familial predisposition for ovarian cancer (OR: 0.22; 95% CI: 0.06-0.88; p=0.03). Conclusion: in this study, having comorbidities was associated with a lower risk of having a familial predisposition for ovarian cancer. Other factors associated with the risk of ovarian cancer did not have an impact on this predisposition

Somatic mutations in breast and serous ovarian cancer young patients:a systematic review and meta-analysis

Objective: our aim was to evaluate whether somatic mutations in five genes were associated with an early age at presentation of breast cancer (BC) or serous ovarian cancer (SOC). Methods: COSMIC database was searched for the five most frequent somatic mutations in BC and SOC. A systematic review of PubMed was performed. Young age for BC and SOC patients was set at Results: twenty six (1,980 patients, 111 younger) and 16 studies (598, 41 younger), were analyzed for BC and SOC, respectively. In BC, PIK3CA wild type tumor was associated with early onset, not confirmed in binary regression with estrogen receptor (ER) status. In HER2-negative tumors, there was increased frequency of PIK3CA somatic mutation in older age groups; in ER-positive tumors, there was a trend towards an increased frequency of PIK3CA somatic mutation in older age groups. TP53 somatic mutation was described in 20% of tumors from both younger and older patients; PTEN, CDH1 and GATA3 somatic mutation was investigated only in 16 patients and PTEN mutation was detected in one of them. In SOC, TP53 somatic mutation was rather common, detected in more than 50% of tumors, however, more frequently in older patients. Conclusion: frequency of somatic mutations in specific genes was not associated with early-onset breast cancer. Although very common in patients with serous ovarian cancer diagnosed at all ages, TP53 mutation was more frequently detected in older women

Somatic and germline mutations in young patients with breast cancer

Aproximadamente 4% das pacientes com câncer de mama têm o diagnóstico abaixo dos 36 anos. Mutação germinativa nos genes BRCA1 ou BRCA2 pode ser um dos fatores de predisposição e a identificação de pacientes carreadoras pode permitir o direcionamento do tratamento e o aconselhamento familiar para medidas de prevenção e detecção precoce. Entretanto, a maioria das pacientes apresenta câncer esporádico e fatores predisponentes são menos evidentes. Para estas pacientes, a identificação de mutações somáticas pode permitir a melhor compreensão da biologia da doença e o desenvolvimento de tratamentos dirigidos a alvos moleculares. Assim, nossos objetivos foram avaliar em pacientes jovens com câncer de mama: história familiar, hábitos de vida, mutação germinativa nos genes BRCA1 e BRCA2 e mutações somáticas no tumor. Oitenta e três pacientes diagnosticadas com câncer de mama entre 18-35 anos foram entrevistadas usando um questionário. O DNA foi extraído do sangue periférico e toda região codificadora dos genes BRCA1/2 foi sequenciada pelo método de sequenciamento de Sanger e a pesquisa de grandes deleções e inserções foi realizada por Amplificação Dependente da Ligação de Múltiplas Sondas (MLPA). Os resultados foram analisados utilizando-se os programas Mutation Surveyor v.3.20 e Chromas version 2.13 e as mutações foram caracterizadas utilizando-se os bancos de dados BIC, LOVD, LOVD-IARC, UMD e ClinVar. O sequenciamento completo do exoma foi realizado em amostras de sangue e tumor fresco congelado de oito pacientes (receptor hormonal positivo, HER2 negativo e BRCA1/2 tipo selvagem) usando Nextera Rapid Capture Enrichment e sequenciadas no Illumina HiSeq 1000. Para detecção de alterações somáticas provenientes do sequenciamento completo do exoma, foi utilizado o programa SomaticSniper (v1.0.2). Foi também analisada a presença de mutação somática no gene PIK3CA em amostras tumorais em blocos de parafina de pacientes jovens e pacientes mais idosas com câncer de mama. Além disso, foi realizada uma meta análise para avaliar se mutação somática nos cinco genes mais frequentemente mutados em câncer de mama estão associados à idade precoce. A idade mediana das 83 pacientes ao diagnóstico foi 32 anos (22-35 anos); idade mediana da menarca foi 13 anos (8-19 anos); 71,1% já passaram por pelo menos uma gestação a termo com idade mediana de 22 anos (14-34 anos); 80,7% nunca fumaram; 74,7% não ingerem bebida alcoólica regularmente; a média para índice de massa corpórea foi 25,26 (10,78-41,57). A maioria das pacientes teve diagnóstico de carcinoma ductal invasivo (89,3%), grau histológico III (49,4%), subtipo luminal (65,9%) e com expressão de Ki67 >14% (89,2%). Aproximadamente 52% das pacientes relataram história familiar para câncer de mama, ovário ou próstata. Mutações deletérias nos genes BRCA1/2 foram detectadas em 13 pacientes (BRCA1, n= 4 e BRCA2, n= 9). Uma mutação nova, que gera um códon de terminação (c.483T>A; p.Cys161Ter), foi detectada no exon 6 do gene BRCA2. O sequenciamento do exoma foi realizado em amostras de oito pacientes carreadoras de BRCA1/2 tipo selvagem e tumor subtipo luminal e revelou uma média de 39 alterações somáticas por tumor (variando entre 19-74). Foram observadas alterações com potencial driver em 53 genes, como PIK3CA, PRKD1, PRKAR1A e PIK3AP1, que codificam proteínas tirosina quinase ou proteínas envolvidas na regulação de proteínas quinases; e no gene POLD1, que codifica a subunidade catalítica da polimerase delta, com papel na replicação e reparo do DNA. Em uma meta análise observamos que a mutação somática no gene PIK3CA é relativamente frequente em pacientes jovens e idosas. Concluindo, mais de 15% das pacientes jovens são carreadoras de mutação deletéria nos genes BRCA1 ou BRCA2. Em pacientes com câncer esporádico foram encontradas mutações somáticas em genes que codificam proteínas tirosina quinase ou proteínas envolvidas em reparo de DNA. Em consonância com o observado em tumores de pacientes mais idosas, mutação somática no gene PIK3CA é relativamente frequente em tumores de pacientes jovensApproximately 4% of the breast cancer patients have their diagnosis under the age of 36 years and germline mutations in BRCA1 or BRCA2 genes is one of the predisposing factors. Identification of patients who are mutation carriers may contribute for the adoption of targeted therapies and for genetic counseling of their family members for early detection or preventive measures. However, most patients present with sporadic cancer where predisposing factors are less understood. In the latter group of patients, the identification of somatic mutations may contribute to a better understanding of the biology of the disease and to the development of molecular targeted therapies. Therefore, our goals were to characterize early onset breast cancer patients for family history, lifestyle, germline mutations in BRCA1 and BRCA2 genes and somatic mutations. Eighty-three breast cancer patients aged 18-35 years were interviewed using a questionnaire. DNA was extracted from peripheral blood and all coding regions of BRCA1/2 genes were screened by Sanger sequencing and large deletions and insertions were examined by Multiplex Ligation-Dependent Probe Amplification (MLPA). Results were analyzed through Mutation Surveyor v.3.20 and Chromas version 2.13 and searched in BIC Database, LOVD, LOVD-IARC, UMD and ClinVar. Whole exome sequencing was performed in blood and fresh-frozen tumor samples from eight patients (hormone receptor positive, HER2 negative and BRCA1/2 wild type) using Nextera Rapid Capture Enrichment in an Illumina HiSeq 1000. To detect somatic SNVs from whole exome sequencing data, SomaticSniper (v1.0.2) was used. Somatic mutation in PIK3CA gene was then searched in Formaldehyde Fixed-Paraffin Embedded samples from another group of young and older breast cancer patients. In addition, a metaanalysis was performed to evaluate whether somatic mutations in the five genes most frequently mutated in breast cancer patients were associated with an early age onset. Median age of 83 patients at diagnosis was 32 years (22-35y), median age of menarche was 13 years (8-19y); 71.1% patients had at least one born child, median age of first pregnancy was 22 years (14-34y); 80.7% were never smokers; 74.7% reported no regular alcoholic drinking; median body mass index was 25.26 (10.78-41.57). Most patients presented with invasive ductal carcinoma (89.3%), histological grade III (49.4%), luminal subtype (65.9%) and Ki67 expression > 14% (89.2). Approximately 52% of the patients reported a positive family history for breast, ovarian or prostate cancer. Deleterious mutations in BRCA1/2 genes ware detected in 13 patients (BRCA1, n= 4 and BRCA2, n= 9). One novel mutation was detected in BRCA1 gene: a stop codon in exon 6 (c.483T > A; p.Cys161Ter). Exome sequencing was evaluated in eight luminal tumors fromBRCA1/2 wild type patients and a median of 39 somatic alterations/tumor was detected, varying from 17 to 74. Potential driver alterations were detected in 53 genes, such as PIK3CA, PRKD1, PRKAR1A and PIK3AP1, that encode tyrosine kinase proteins or proteins involved in tyrosine kinases regulation; and POLD1 gene, that encodes the catalytic subunit of DNA polymerase delta which plays a critical role in DNA replication and repair. A Meta-analysis showed that somatic mutation in PIK3CA gene was frequently detected in both young and older patients. In conclusion, more than 15% of the young breast cancer patients are BRCA1 or BRCA2 mutation carriers. In patients with sporadic cancer somatic mutations were found in genes that encode tyrosine kinase proteins or are involved in the DNA repair. In agreement with what was observed in tumors from older patients, somatic mutation in PIK3CA gene is relatively frequent in tumors from young patient

Detection of Markers for Germ Cells in Rats during the Embryonic Phase: Differences and Similarities with Mice

Germ cells are the only cells capable of transmitting genetic information from generation to generation. Germ cell development has been widely studied in different species. Among mammals, the mouse is the model used in the majority of the studies on Primordial Germ Cell (PGC) differentiation, sex-determination and genetics. The results obtained in the present study show that the rat is also a very important model for studies about germ cell development. For this study, PGC, gonocyte and Sertoli cell markers were used such as: GCNA1, MVH, SSEA1 and OCT4 to germ cells and GATA4 to Sertoli cells. The choice of theses markers was based in the studies performed with mice, once there is not enough available information about the expression of germ cell markers in the rat during the embryonic phase. The presence these markers was investigated through immunofluorescence and immunohistochemistry. The results showed that SSEA1, GCNA1 and MVH proteins show very distinct labeling pattern in mice and rats. SSEA1 and GCNA1 proteins were undetectable in the PGC. However MVH was detected in migrating PGC and in those that had already colonized the gonad. A similar labeling pattern was also observed in humans, suggesting some degree of similarity between these two species. The transcription factor OCT4 showed similar labeling to that described in mice. However, we observed that OCT4 is not detected in germ cells from dos 19dpc on. The results obtained in this study are of extreme relevance, since important differences between the rats and mice germ cells were shown. These results show the importance of including the rat as an experimental model in the studies about germ cells development.As células germinativas são as únicas células capazes de transmitir a informação genética de geração em geração. O desenvolvimento dessas células tem sido largamente estudado em diferentes espécies. Dentre os mamíferos, o camundongo é o modelo usado na maioria dos estudos sobre diferenciação, determinação sexual e genética das Células Germinativas Primordiais (CGP). Os dados obtidos no presente estudo sugerem que o rato também é um importante modelo para os estudos sobre o desenvolvimento das células germinativas. Para este estudo, foram utilizados alguns marcadores de CGP, gonócitos e de células de Sertoli, tais como: GCNA1, MVH, SSEA1 e OCT4 para as células germinativas e GATA4 para as células de Sertoli. A escolha desses marcadores se deu com base nos estudos realizados em camundongo, uma vez que não há informação suficiente sobre a expressão de marcadores de células germinativas em ratos durante a fase embrionária. A presença desses marcadores foi investigada através de imunofluorescência e imunohistoquímica. Os resultados mostraram que as proteínas SSEA1, GCNA e MVH apresentam padrão de marcação bastante diferente daquele observado em camundongos, sugerindo que talvez estas proteínas tenham função diferente no rato. As proteínas SSEA1 e GCNA1 não foram detectadas nas CGP, enquanto a proteína MVH foi detectada tanto nas CGP em migração quanto naquelas que já colonizaram as gônadas. Este mesmo padrão de marcação foi observado em humanos, sugerindo semelhanças entre essas duas espécies. Nas idades estudadas, o fator de transcrição OCT4 apresentou marcação semelhante àquela descrita em camundongos. Entretanto, o OCT4 não é mais detectado nas células germinativas a partir dos 19dpc. Os dados obtidos neste estudo sugerem a existência de importantes diferenças na expressão de marcadores das células germinativas entre ratos e camundongos, demonstrando a importância de se incluir o rato como modelo experimental nos estudos sobre desenvolvimento das células germinativas.TEDEBV UNIFESP: Teses e dissertaçõe

Detection of Four Germ Cell Markers in Rats during Testis Morphogenesis: Differences and Similarities with Mice

Germ cells are the only cells capable of transmitting genetic information from generation to generation. Germ cell development has been widely studied in different species. Among mammals, the mouse is the model used in the majority of studies on germ cell differentiation, sex determination and genetics. in the present study, we suggest that the rat is also a very important model for the investigation of the mechanisms of germ cell development. To study rat germ cell development and compare it with that of mouse, the germ cell markers germ cell nuclear antigen 1 (GCNA1), OCT4, mouse vasa homologue (MVH) and specific surface embryonic antigen 1 (SSEA1) were immunolabeled at different phases of embryonic and postnatal development. SSEA1 and GCNA1 were not detected in rat primordial germ cells and fetal gonocytes. GCNA1 was detected postnatally and was present only in leptotene, zygotene and early pachytene spermatocytes. On the other hand, in mice, these markers were detected in germ cells as soon as 11.5 days postcoitum (dpc). MVH was detected in migrating rat primordial germ cells as well as in those that have already colonized the gonads, whereas in mice, MVH is detected only in germ cells that have reached the gonads. in rats, OCT4-positive germ cells were detected from 13 to 17 dpc, but not at 19 dpc or in postnatal testes. This is in contrast with mice that show OCT4 labeling in both embryonic and adult testes. These data suggest that primordial germ cell development in rats and mice shows considerable differences and that the rat may also be an important model to study the embryonic development of germ cells. Copyright (C) 2011 S. Karger AG, BaselFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fed Univ São Paulo UNIFESP, Dept Morphol & Genet, Dev Biol Lab, BR-04023900 São Paulo, BrazilFed Univ São Paulo UNIFESP, Dept Morphol & Genet, Dev Biol Lab, BR-04023900 São Paulo, BrazilFAPESP: 2009/07170-3, 2009Web of Scienc

Additional file 1: of Germline mutations in BRCA1 and BRCA2 in epithelial ovarian cancer patients in Brazil

Supplementary Methods. Table S1. Clinical and pathological characteristics, BRCA sequencing and MLPA results. Table S2. BRCA1 gene variants. Table S3. BRCA2 gene variants. (ZIP 73.7 kb