RESUMO
Breast cancer and ovarian cancer pose a significant risk for BRCA1 carriers, with limited risk-reduction strategies. While improved screening helps in the early detection of breast cancer, preventive measures remain elusive. Emerging evidence suggests a potential link between iodine levels and modulation of cancer risk, but comprehensive studies are scarce. We conducted a prospective study among 989 BRCA1 carriers to assess the association between blood iodine levels and breast and ovarian cancer risk. Using inductively coupled plasma mass spectrometry, we measured blood iodine levels and observed a negative association with breast cancer risk, with a significantly lower risk observed in quartile 4 (iodine > 38.0 µg/L) compared with quartile 1 (iodine < 30 µg/L) (HR = 0.49; 95%CI: 0.27-0.87; p = 0.01). Conversely, a suggestive increase in ovarian cancer risk was observed at higher iodine levels (HR = 1.91; 95%CI: 0.64-5.67; p = 0.25). No significant association was found between iodine levels and overall cancer risk. Our results suggest the potential of iodine to reduce breast cancer risk in BRCA1 carriers after prophylactic oophorectomy but require further validation and investigation of its effect on ovarian cancer risk and overall mortality. These findings highlight the need for personalized strategies to manage cancer risk in BRCA1 carriers.
Assuntos
Proteína BRCA1 , Neoplasias da Mama , Iodo , Neoplasias Ovarianas , Humanos , Feminino , Neoplasias da Mama/sangue , Neoplasias da Mama/genética , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/genética , Iodo/sangue , Pessoa de Meia-Idade , Adulto , Estudos Prospectivos , Proteína BRCA1/genética , Proteína BRCA1/sangue , Fatores de Risco , Heterozigoto , Biomarcadores Tumorais/sangue , IdosoRESUMO
Recently, our lab, part of a referral center in Italy, reported its experience regarding the execution of germline BRCA1/2 (gBRCA) testing during the first months of the coronavirus disease-2019 (COVID-19) pandemic, which highlights a substantial reduction (about 60%) compared with the first 2 months of the current year. This evidence appeared to be a lockdown effect due to extraordinary restriction measures to slow down the spread of SARS-CoV-2. In this study, we aimed to evaluate the overall effects of the ongoing pandemic on gBRCA testing in our institution and to understand how COVID-19 has influenced testing after the complete lockdown (March 8-May 5, 2020). Additionally, we compared this year's trend with trends of the last 3 years to better monitor gBRCA testing progress. This detailed analysis highlights two important findings: (1) gBRCA testing did not increase significantly after the lockdown period (May-October 2020) compared with the lockdown period (March-April 2020), emphasizing that even after the lockdown period testing remained low. (2) Comparing the total tests per year (January-October 2017, 2018, 2019, with 2020), the impact of COVID-19 on gBRCA testing is apparent, with similarities of trends registered in 2017. These evidences reveal a gBRCA testing delay for cancer patients and healthy patients at this moment, and the new era of gBRCA testing in the management of ovarian, breast, pancreas and prostate cancer patients has been seriously questioned due to the COVID-19 pandemic. As consequence, we underline that measures to guarantee oncogenetic testing (e.g., gBRCA testing) along with new diagnostic/clinic strategies are mandatory. For these reasons, several proposals are presented in this study.
Assuntos
Proteína BRCA1/sangue , Neoplasias da Mama/diagnóstico , COVID-19/epidemiologia , Neoplasias Ovarianas/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Pandemias , Neoplasias da Próstata/diagnóstico , Biomarcadores Tumorais/sangue , COVID-19/psicologia , Diagnóstico Tardio/ética , Detecção Precoce de Câncer/estatística & dados numéricos , Feminino , Política de Saúde , Humanos , Itália/epidemiologia , Masculino , Distanciamento Físico , Quarentena/psicologia , SARS-CoV-2/patogenicidadeRESUMO
Background and objective: BRCA1 and BRCA2 are associated with many cancer types in addition to hereditary breast and ovarian cancers. However, their relation to lung cancer remains to be explored. Materials and Methods: Observation studies were systematically reviewed to explore the association of BRCA1 or BRCA2 with lung cancer. PubMed, MEDLINE [EBSCOhost], and relevant articles published up to 7 January 2020 were searched. Odd ratio (OR), standardized morbidity rate (SMR), and cancer-specific standardized incidence ratios (SIRs) were pooled together as relative risk (RR) estimates (95% confidence interval [CI], 0.66-1.40). Results: Thirteen studies were included for analysis. Results showed that the RR of BRCA2 is 0.76 (95% CI, 0.48-1.19), the overall RR is 0.96 (95% CI, 0.66-1.40), and that of BRCA1 is 0.66 (95% CI, 0.41-1.05), indicating that it was not associated with lung cancer. Conclusion: With the limitation of the retrospective study design and severe heterogeneity, these results inform clinicians and relevant families that BRCA1 and BRCA2 mutation carriers have no increased risk of lung cancer.
Assuntos
Proteína BRCA1/análise , Proteína BRCA2/análise , Neoplasias Pulmonares/sangue , Adulto , Idoso , Proteína BRCA1/sangue , Proteína BRCA2/sangue , Feminino , Predisposição Genética para Doença , Humanos , Incidência , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Razão de Chances , Estudos RetrospectivosRESUMO
OBJECTIVES: Breast cancer susceptibility gene 1/2 (BRCA1/2) is a promising tumor marker in many types of cancer. However, the methylation frequency of BRCA1/2 gene with occurrence risk and survival benefit of patients with breast carcinoma remains controversy. The aim of the present study was to assess the relationship between BRCA1/2 gene promoter methylation and the occurrence and prognosis in breast carcinoma based on a meta-analysis, meanwhile, this article explored the differential expression levels of BRCA1/2 gene promoter methylation in peripheral blood and tumor tissues of breast cancer patients. METHODS: Electronic databases (PubMed, Medline, Cochrane Library, and CNKI) were searched up to June 2019. The number of BRCA1/2 promoter methylation-positive and -negative patients in breast carcinoma patients were measured, and hazard ratio (HR) with 95% confidence interval (CI) for the association between BRCA1/2 gene promoter methylation and the prognosis of breast carcinoma patients. Primary end points were presence of breast cancer, overall survival (OS), disease-free survival (DFS). Statistical analysis was performed with STATA 12.0. RESULTS AND CONCLUSIONS: Fifty-eight articles including 19,084 individuals met full eligibility criteria. We observed that the frequency of BRCA1 gene promoter methylation was higher in breast cancer tissues compared with normal tissues, and the prognostic analysis suggested that BRCA1 gene promoter methylation was significantly associated with poor overall survival and poor disease-free survival. This study also verified that there was no statistically significant difference in the methylation frequency of BRCA1 gene promoter between peripheral blood and tumor tissues in breast cancer patients, which suggests that the detection of BRCA1 promoter methylation in peripheral blood may be a non-invasive and rapid way to monitor the occurrence breast cancer.
Assuntos
Proteína BRCA1/análise , Proteína BRCA2/análise , Neoplasias da Mama/genética , Metilação de DNA/genética , Prognóstico , Proteína BRCA1/sangue , Proteína BRCA2/sangue , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/sangue , Neoplasias da Mama/fisiopatologia , Feminino , Humanos , Pessoa de Meia-Idade , Modelos de Riscos ProporcionaisRESUMO
Correct classification of genomic variants causing potentially aberrant splicing is of utmost importance for patient management, especially in clinically actionable genes such as BRCA1/2. In this article, we report molecular evaluation of the BRCA1 c.439T>C (rs794727800, p.Leu147=) variant based on RNA of a patient suffering with high-grade serous ovarian cancer syndrome, to add new evidence to the only in silico data available for this variant. High Resolution Melting Analysis (HRMA) was used for the first time to investigate the spliceogenicity of a BRCA1 variant. HRMA with Sanger sequencing provided evidence that the c.439C allele does not cause aberrant splicing of the BRCA1 exon 7. In addition, HRMA with Sanger highlighted a different expression of the naturally occurring BRCA1 r.442_444del (c.442_444delCAG, p.Gln148del, at DNA level) isoform between blood and tumor, in this patient. HRMA is an alternative molecular approach to analyze spliceogenic properties of the c.439T>C variant and potentially for all those BRCA1/2 variants affecting splicing sites. These new evidences allowed to classify definitively the c.439T>C variant as benign. Furthermore, the different BRCA1 r.442_444del expression opens the discussion to consider a wider classification criteria for the splicing variants, including molecular evaluation at tissue level, which is an aspect currently scarcely considered in BRCA1/2 variant classification recommendations.
Assuntos
Proteína BRCA1/genética , Desnaturação de Ácido Nucleico/genética , Polimorfismo de Nucleotídeo Único/genética , Splicing de RNA/genética , Idoso , Alelos , Proteína BRCA1/sangue , Sequência de Bases , DNA/genética , Feminino , HumanosRESUMO
OBJECTIVE: Ovarian cancer is one of the leading causes of cancer deaths in women. Ovarian cancer is diagnosed at the late stages and generally relapses within 12-14 months of cytoreductive surgery. This is attributed to lack of precise molecular detection methodologies to detect and track the disease. Epigenetic alteration such as aberrant promoter hypermethylation is an important early event that occurs during cancer development and progression. This study focuses on development of a minimally invasive methylation marker that could be used for detection and prognosis of ovarian cancer patients. METHODS: Aberrant promoter hypermethylation of RASSF1a and BRCA1 was assessed in circulating DNA of 72 EOC patients using methylation-specific PCR. The findings were correlated with various clinicopathological parameters. Statistical analysis was done using the Fisher exact test and chi-square test. RESULTS: The aberrant methylation patterns of RASSF1a and BRCA1 was identified to be present in the cancerous samples. A total of 31.9 % and 56.9% methylation was observed for RASSF1a and BRCA1 respectively. A striking 50% methylation of BRCA1 was identified in the benign sample cohort, which marks the significance of assessing the hypermethylation pattern to detect cancer at its early stages. Methylation of the two tumor suppressor genes was evident across various stages and grades of ovarian tumors suggesting that this could also help as a prognostic marker. CONCLUSION: The results of the current study hold significance since the hypermethylation patterns can be identified in the cell-free circulating tumor DNA from a small volume of blood plasma and is a simple and minimally-invasive method. Assessment of hypermethylation patterns of a panel of TSG along with the existing screening markers could aid in better diagnosis and management of the disease. It could also aid in designing specifically tailored treatment strategies to fight the disease.
Assuntos
Proteína BRCA1/genética , Biomarcadores Tumorais/genética , DNA Tumoral Circulante/genética , Metilação de DNA , Neoplasias Ovarianas/patologia , Regiões Promotoras Genéticas , Proteínas Supressoras de Tumor/genética , Adenocarcinoma de Células Claras/sangue , Adenocarcinoma de Células Claras/genética , Adenocarcinoma de Células Claras/patologia , Adenocarcinoma Mucinoso/sangue , Adenocarcinoma Mucinoso/genética , Adenocarcinoma Mucinoso/patologia , Proteína BRCA1/sangue , Biomarcadores Tumorais/sangue , Estudos de Casos e Controles , DNA Tumoral Circulante/sangue , Cistadenocarcinoma Seroso/sangue , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/patologia , Neoplasias do Endométrio/sangue , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Feminino , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/genética , Prognóstico , Proteínas Supressoras de Tumor/sangueRESUMO
Women carrying a BRCA mutation have an increased risk of developing breast and ovarian cancer. The most effective strategy to reduce this risk is the bilateral salpingo-oophorectomy, with or without additional risk-reducing mastectomy. Risk-reducing bilateral salpingo-oophorectomy (RRBSO) is recommended between age 35 and 40 and between age 40 and 45 years for women carriers of BRCA1 and BRCA2 mutations, respectively. Consequently, most BRCA mutation carriers undergo this procedure prior to a natural menopause and develop an anticipated lack of hormones. This condition has a detrimental impact on various systems, affecting both the quality of life and longevity; in particular, women carrying BRCA1 mutation, who are likely to have surgery earlier as compared to BRCA2. Hormonal replacement therapy (HRT) is the only effective strategy able to significantly compensate the hormonal deprivation and counteract menopausal symptoms, both in spontaneous and surgical menopause. Although recent evidence suggests that HRT does not diminish the protective effect of RRBSO in BRCA mutation carriers, concerns regarding the safety of estrogen and progesterone intake reduce the use in this setting. Furthermore, there is strong data demonstrating that the use of estrogen alone after RRBSO does not increase the risk of breast cancer among women with a BRCA1 mutation. The additional progesterone intake, mandatory for the protection of the endometrium during HRT, warrants further studies. However, when hysterectomy is performed at the time of RRBSO, the indication of progesterone addition decays and consequently its potential effect on breast cancer risk. Similarly, in patients conserving the uterus but undergoing risk-reducing mastectomy, the addition of progesterone should not raise significant concerns for breast cancer risk anymore. Therefore, BRCA mutation carriers require careful counselling about the scenarios following their RRBSO, menopausal symptoms or the fear associated with HRT use.
Assuntos
Terapia de Reposição Hormonal/métodos , Salpingo-Ooforectomia/métodos , Adulto , Proteína BRCA1/análise , Proteína BRCA1/sangue , Proteína BRCA2/análise , Proteína BRCA2/sangue , Feminino , Predisposição Genética para Doença/genética , Predisposição Genética para Doença/prevenção & controle , Terapia de Reposição Hormonal/normas , Humanos , Pessoa de Meia-Idade , Comportamento de Redução do Risco , Salpingo-Ooforectomia/reabilitaçãoRESUMO
Male breast cancer (MBC) is rare and consequently understudied. Here we present the case of contralateral breast cancer in a male patient nearly a quarter century following his initial breast cancer diagnosis and treatment. The epidemiology, risk factors, diagnosis, characterization, treatment, and prognosis of male breast cancer are reviewed. MBC accounts for <1% of all breast cancer with an estimated incidence nearly 1.25 per 100,000 person years. Our patient tested positive for ATM mutation of undetermined significance. More commonly in males, a BRCA2 mutation confers a >1 in 15 lifetime risk of breast cancer and is present in >11% of MBC patients, while BRCA1 is present in an estimated <1.5% of MBC patients. The risk of contralateral breast cancer developing in a male with a unilateral lesion is much higher than for a primary male breast cancer in the general population. Men tend to be diagnosed at a later age and stage than females. Prognosis for male and female breast cancer is similar considering both age of patient and stage of the tumor at diagnosis, and similar treatment paradigms have resulted in similar outcomes. Although lumpectomy with radiation therapy may have the same prognosis as mastectomy, the standard of care for male breast cancer continues to be simple mastectomy with sentinel lymph node biopsy.
Assuntos
Neoplasias da Mama Masculina/diagnóstico , Idoso , Proteína BRCA1/análise , Proteína BRCA1/sangue , Neoplasias da Mama Masculina/diagnóstico por imagem , Neoplasias da Mama Masculina/cirurgia , Angiografia por Tomografia Computadorizada/métodos , Ginecomastia/diagnóstico , Ginecomastia/etiologia , Humanos , Incidência , Masculino , Mastectomia/métodos , Fatores de RiscoRESUMO
INTRODUCTION: The purpose of this study was to identify previvors' strategies for communicating about family planning after testing positive for a variant of the "breast cancer gene" (BRCA). METHOD: Semistructured interviews were conducted with 20 women currently in committed romantic relationships, but who had not yet completed family planning upon finding out about their BRCA mutation status. RESULTS: Data analysis produced three categories of participant advice given to newly diagnosed previvors. Participants advised the following: (a) the importance of engaging in two-way dialogue with their partners/spouses across the life span of the partnership, (b) seeking information on new technologies and information regarding family-planning and genetic-cancer-prevention decision-making, as well as recognizing where to go for different support needs, and (c) managing and acknowledging emotions surrounding their BRCA-related health decisions. DISCUSSION: Previvors who have already had family-planning and genetic-cancer-risk conversations had important advice for newly diagnosed previvors. Practical advice for starting and managing conversations with partners/spouses, family members, and friends are discussed. (PsycINFO Database Record
Assuntos
Neoplasias da Mama/psicologia , Comunicação , Serviços de Planejamento Familiar/métodos , Predisposição Genética para Doença/psicologia , Adulto , Proteína BRCA1/análise , Proteína BRCA1/sangue , Proteína BRCA2/análise , Proteína BRCA2/sangue , Neoplasias da Mama/diagnóstico , Detecção Precoce de Câncer/psicologia , Serviços de Planejamento Familiar/tendências , Feminino , Humanos , Pessoa de Meia-Idade , Pesquisa Qualitativa , Fatores de RiscoRESUMO
AIMS: In this study, we determined whether serum tumor markers (STMs), including CA125, are associated with BRCA mutation status and if they can be used prognostically in sporadic ovarian cancer (SOC) and familial ovarian cancer (FOC). METHODS: BRCA gene mutations were screened using next-generation sequencing (NGS) in 31 FOC and 66 SOC patients enrolled between 2013 and 2014. The serum levels of STM CEA, CA125, CA199, and HE4 were also measured in these patients to determine the prognostic potential of these markers and their association with BRCA mutations. RESULTS: Elevated levels of CA125, but not the other three STMs, were associated with FOC and BRCA mutations. Median progression-free survival (PFS) was significantly longer in patients with FOC, higher CA125 expression (>2000 U/mL), and BRCA mutation. Strikingly, the median PFS was not reached in either BRCA+/higher CA125 or FOC/higher CA125 patient groups and these patients had significantly longer PFS than those in other groups. As reported previously, we also detected more BRCA mutations in FOC than in SOC. No significant differences were observed in onset age, menopausal status, tumor stage, and distant metastasis between FOC and SOC patients or between BRCA+ and BRCA- patients. CONCLUSIONS: Elevated levels of serum CA125 are associated with FOC and BRCA mutations, which can be further exploited as a prognostic marker in OC.
Assuntos
Proteína BRCA1/genética , Proteína BRCA2/genética , Antígeno Ca-125/sangue , Proteínas de Membrana/sangue , Neoplasias Ovarianas/genética , Adulto , Idoso , Proteína BRCA1/sangue , Proteína BRCA1/metabolismo , Proteína BRCA2/sangue , Proteína BRCA2/metabolismo , Biomarcadores Tumorais/sangue , Antígeno Ca-125/metabolismo , Intervalo Livre de Doença , Feminino , Humanos , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Mutação , PrognósticoRESUMO
Genome-wide Illumina InfiniumMethylation 450 K DNA methylation analysis was performed on blood samples from clinical atherosclerosis patients (n = 8) and healthy donors (n = 8) in the LVAD study (NCT02174133, NCT01799005). Multiple differentially methylated regions (DMR) could be identified in atherosclerosis patients, related to epigenetic control of cell adhesion, chemotaxis, cytoskeletal reorganisations, cell proliferation, cell death, estrogen receptor pathways and phagocytic immune responses. Furthermore, a subset of 34 DMRs related to impaired oxidative stress, DNA repair, and inflammatory pathways could be replicated in an independent cohort study of donor-matched healthy and atherosclerotic human aorta tissue (n = 15) and human carotid plaque samples (n = 19). Upon integrated network analysis, BRCA1 and CRISP2 DMRs were identified as most central disease-associated DNA methylation biomarkers. Differentially methylated BRCA1 and CRISP2 regions were verified by MassARRAY Epityper and pyrosequencing assays and could be further replicated in blood, aorta tissue and carotid plaque material of atherosclerosis patients. Moreover, methylation changes at BRCA1 and CRISP2 specific CpG sites were consistently associated with subclinical atherosclerosis measures (coronary calcium score and carotid intima media thickness) in an independent sample cohort of middle-aged men with subclinical cardiovascular disease in the Aragon Workers' Health Study (n = 24). Altogether, BRCA1 and CRISP2 DMRs hold promise as novel blood surrogate markers for early risk stratification and CVD prevention.
Assuntos
Aterosclerose/genética , Proteína BRCA1/genética , Biomarcadores/sangue , Metilação de DNA , Glicoproteínas/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Aterosclerose/sangue , Proteína BRCA1/sangue , Espessura Intima-Media Carotídea , Moléculas de Adesão Celular , Estudos de Coortes , Ilhas de CpG , Epigênese Genética , Feminino , Redes Reguladoras de Genes , Glicoproteínas/sangue , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Sequenciamento Completo do GenomaRESUMO
In this study, we report an enzyme-free and conjugation-free electrochemical genosensor enabling an ultrasensitive readout of BRCA-1, a breast cancer susceptibility gene. The sensor employs a target-responsive hybridization chain reaction (HCR) to significantly amplify the detectable current signals. By means of a functional auxiliary probe pair and a versatile initiator sequence, a linear DNA concatemer structure can be formed via spontaneous and continuous polymerization of DNA oligomers in the presence of target sequence. Such a DNA nanoassembly endows the genosensor an ultrahigh sensitivity up to 1 aM, which is higher than that of the nanomaterials-based or enzyme mediated amplification approaches by several orders of magnitude. More importantly, the sensor's responsive peak current exhibits a favorable linear correlation to the logarithm of the concentrations of target sequence ranging from 1 aM to 10 pM. In addition, the sensor is highly selective, and can discriminate a single mismatched sequence. This HCR-based genosensor is also capable of probing low-abundance BRCA-1 gene sequence directly in complex matrices, such as 50% human serum, with minimal interference. These advantages will make our tailor-engineered HCR-based electrochemical genosensor appealing to genetic analysis and clinical diagnostics.
Assuntos
Proteína BRCA1/sangue , Proteína BRCA1/isolamento & purificação , Técnicas Biossensoriais , Neoplasias da Mama/sangue , Proteína BRCA1/genética , Neoplasias da Mama/genética , Feminino , Humanos , Hibridização In SituRESUMO
Breast cancer is a most common malignancy especially in Iraqi women accounting for high morbidity and mortality. Mutations in BRCA1 gene is one of the important genetic predisposing factors inbreast cancer. Similarly ERBB2 and TP53 are also key prognostic markers in breast cancer treatment.We were interested to explore the gene expression profiles of BRCA1, ERBB2 and TP53 in breast cancer women patients from Iraq so as to assess the potential of such markers in breast cancer treatment. The mRNA levels were significantly over-expressed in tumor tissues in comparison to normal ones with p values (p<0.005) observed between malignant BRCA1 and control tissue samples. Similarly significant difference (p<0.001) was observed between malignant ERBB2 in comparison to control, and malignant TP53 and benign tissue samples as well. However in blood samples, no considerable expression of these markers was observed. Out of three selected genes, ERBB2 expression was significantly expressed in comparison to BRCA1 and TP53 in cancer tissue. Mutation analysis of BRCA1, ERBB2 and TP53 has been made to find out the region most susceptible to mutations in these genes The BRCA1 exon 11, ERBB2 16 and TP53 exon 5 displayed increased chances of having mutations. We can conclude from the study that differential gene expression of BRCA1, ERBB2 and TP53 at mRNA levels may act as a diagnostic marker of circulating tumor cells having important prognostic value in breast cancer patients.
Assuntos
Proteína BRCA1 , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Neoplasias da Mama/sangue , Neoplasias da Mama/genética , Regulação Neoplásica da Expressão Gênica , Receptor ErbB-2 , Proteína Supressora de Tumor p53 , Proteína BRCA1/sangue , Proteína BRCA1/genética , Neoplasias da Mama/patologia , Feminino , Perfilação da Expressão Gênica , Humanos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptor ErbB-2/sangue , Receptor ErbB-2/genética , Proteína Supressora de Tumor p53/sangue , Proteína Supressora de Tumor p53/genéticaRESUMO
A label-free and low fouling biosensor based on functional polyethylene glycols selective for breast cancer susceptibility gene (BRCA1) is reported. Sensory interfaces were prepared through the modification of a glassy carbon electrode with highly cross-linked polyethylene glycol (PEG) film containing amine groups, followed by the self-assembly of gold nanoparticles and the immobilization of BRCA1 complementary single-strand 19-mer oligonucleotides. In the presence of a specific BRCA1 sequence capture and hybridization results in interfacial change sensitively monitored using electrochemical impedance spectroscopy. The combined utilization of a PEG polymer film and gold nanoparticle mixed interface enables very high levels of sensitivity and a highly effective assaying in patient samples. Assay linear range was from 50.0 fM to 1.0 nM, with a limit of detection of 1.72 fM. Furthermore, this label-free DNA sensor has been used for assaying BRCA1 in serum samples, showing its feasible potential for diagnostic applications in clinical analysis of breast cancer gene BRCA1. Foreseeable, this sensor made on this basis undoubtedly provide the most effective and sensitive detection for BRCA1.
Assuntos
Neoplasias da Mama/genética , DNA/genética , Genes BRCA1 , Ouro/química , Nanopartículas Metálicas/química , Polietilenoglicóis/química , Proteína BRCA1/sangue , Proteína BRCA1/genética , Sequência de Bases , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Técnicas Biossensoriais/métodos , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/sangue , Neoplasias da Mama/diagnóstico , DNA/sangue , Técnicas Eletroquímicas/métodos , Eletrodos , Feminino , Humanos , Nanopartículas Metálicas/ultraestrutura , Hibridização de Ácido Nucleico/métodos , Reprodutibilidade dos TestesRESUMO
It has been proposed that methylation signatures in blood-derived DNA may correlate with cancer risk. In this study, we evaluated whether methylation of the promoter region of the BRCA1 gene detectable in DNA from peripheral blood cells is a risk factor for breast cancer, in particular for tumors with pathologic features characteristic for cancers with BRCA1 gene mutations. We conducted a case-control study of 66 breast cancer cases and 36 unaffected controls. Cases were triple-negative or of medullary histology, or both; 30 carried a constitutional BRCA1 mutation and 36 did not carry a mutation. Blood for DNA methylation analysis was taken within three months of diagnosis. Methylation of the promoter of the BRCA1 gene was measured in cases and controls using methylation-sensitive high-resolution melting (MS-HRM). A sample with any detectable level of methylation was considered to be positive. Methylation of the BRCA1 promoter was detected in 15 of 66 cases and in 2 of 36 controls (OR 5.0, p = 0.03). Methylation was present in 15 of 36 women with breast cancer and without germline BRCA1 mutation, but in none of 30 women with breast cancer and a germline mutation (p < 0.01). The association between methylation and breast cancer was restricted to women with no constitutional BRCA1 mutation (OR 12.1, p = 0.0006). Methylation of the promoter of the BRCA1 gene detectable in peripheral blood DNA may be a marker of increased susceptibility to triple-negative or medullary breast cancer.
Assuntos
Proteína BRCA1/genética , Carcinoma Medular/genética , Metilação de DNA/genética , Neoplasias de Mama Triplo Negativas/genética , Adulto , Proteína BRCA1/sangue , Carcinoma Medular/sangue , Carcinoma Medular/patologia , Estudos de Casos e Controles , DNA-Citosina Metilases/genética , Feminino , Mutação em Linhagem Germinativa , Humanos , Regiões Promotoras Genéticas , Neoplasias de Mama Triplo Negativas/sangue , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
Targeted measurements of low abundance proteins in complex mixtures are in high demand in many areas, not the least in clinical applications measuring biomarkers. We here present the novel platform AFFIRM (AFFInity sRM) that utilizes the power of antibody fragments (scFv) to efficiently enrich for target proteins from a complex background and the exquisite specificity of SRM-MS based detection. To demonstrate the ability of AFFIRM, three target proteins of interest were measured in a serum background in single-plexed and multiplexed experiments in a concentration range of 5-1000 ng/mL. Linear responses were demonstrated down to low ng/mL concentrations with high reproducibility. The platform allows for high throughput measurements in 96-well format, and all steps are amendable to automation and scale-up. We believe the use of recombinant antibody technology in combination with SRM MS analysis provides a powerful way to reach sensitivity, specificity, and reproducibility as well as the opportunity to build resources for fast on-demand implementation of novel assays.
Assuntos
Cromatografia Líquida/métodos , Espectrometria de Massas/métodos , Proteoma/metabolismo , Proteômica/métodos , Anticorpos de Cadeia Única/metabolismo , Sequência de Aminoácidos , Afinidade de Anticorpos/imunologia , Proteína BRCA1/sangue , Proteína BRCA1/imunologia , Proteína BRCA1/metabolismo , Humanos , Queratina-19/sangue , Queratina-19/imunologia , Queratina-19/metabolismo , Mucina-1/sangue , Mucina-1/imunologia , Mucina-1/metabolismo , Peptídeos/sangue , Peptídeos/imunologia , Peptídeos/metabolismo , Proteoma/imunologia , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Reprodutibilidade dos Testes , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/imunologiaRESUMO
BACKGROUND: BRCA1 plays an essential role in maintaining genome stability. Inherited BRCA1 germline mutation (BRCA1+) is a determined genetic predisposition leading to high risk of breast cancer. While BRCA1+ induces breast cancer by causing genome instability, most of the knowledge is known about somatic genome instability in breast cancer cells but not germline genome instability. METHODS: Using the exome-sequencing method, we analyzed the genomes of blood cells in a typical BRCA1+ breast cancer family with an exon 13-duplicated founder mutation, including six breast cancer-affected and two breast cancer unaffected members. RESULTS: We identified 23 deleterious mutations in the breast cancer-affected family members, which are absent in the unaffected members. Multiple mutations damaged functionally important and breast cancer-related genes, including transcriptional factor BPTF and FOXP1, ubiquitin ligase CUL4B, phosphorylase kinase PHKG2, and nuclear receptor activator SRA1. Analysis of the mutations between the mothers and daughters shows that most mutations were germline mutation inherited from the ancestor(s) while only a few were somatic mutation generated de novo. CONCLUSION: Our study indicates that BRCA1+ can cause genome instability with both germline and somatic mutations in non-breast cells.
Assuntos
Proteína BRCA1/genética , Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Instabilidade Genômica , Adulto , Proteína BRCA1/sangue , Biomarcadores Tumorais/sangue , Neoplasias da Mama/sangue , Análise Mutacional de DNA , Éxons , Feminino , Efeito Fundador , Predisposição Genética para Doença , Hereditariedade , Humanos , Masculino , Pessoa de Meia-Idade , Mães , Mutação , Núcleo Familiar , Linhagem , FenótipoRESUMO
BACKGROUND: Women with breast cancer increasingly undergo contralateral prophylactic mastectomy (CPM). We evaluated the relationship between preoperative magnetic resonance imaging (MRI) findings and CPM. Other clinicopathologic variables associated with CPM choice and the pathology found in the contralateral breast are also reported. METHODS: Newly diagnosed breast cancer patients were prospectively enrolled in the University of Iowa Breast Molecular Epidemiology Resource. Patients with stages 0-III breast cancer who underwent mastectomy for the index cancer were eligible for this analysis. Univariate logistic regression and a multivariate model were used to identify factors predictive of CPM. RESULTS: Among 134 patients (mean age 54.9 years), 53 (39.6 %) chose CPM. On univariate analysis, patients undergoing CPM were more likely to have a preoperative breast MRI (64.2 vs. 39.5 %, p = 0.006) and to have follow-up testing recommended for the contralateral breast (28.3 vs. 4.9 %, p = 0.001). Univariate analysis also associated CPM with younger age (p < 0.0001), BRCA testing (p < 0.0001), BRCA mutation (p = 0.034) and reconstruction performed (p = 0.001). Median age of youngest child at diagnosis varied significantly between the CPM (15.9 years) and non-CPM (24.3 years) groups (p = 0.0018). On multivariate analysis, MRI follow-up recommendation, young age, reconstruction and human epidermal growth factor receptor 2 (HER2) positivity of the index cancer were significantly associated with CPM. Of the CPM specimens, one (1.8 %) had ductal carcinoma-in situ, which had not been identified on MRI. CONCLUSIONS: Abnormal findings in the contralateral breast on preoperative MRI, as well as young age, reconstruction and HER2-positive status correlated with CPM choice in this cohort. Occult malignancy was rare.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/prevenção & controle , Carcinoma Intraductal não Infiltrante/diagnóstico , Imageamento por Ressonância Magnética , Mastectomia , Cuidados Pré-Operatórios , Prevenção Primária/métodos , Receptor ErbB-2/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteína BRCA1/sangue , Neoplasias da Mama/genética , Neoplasias da Mama/cirurgia , Carcinoma Intraductal não Infiltrante/genética , Carcinoma Intraductal não Infiltrante/cirurgia , Feminino , Seguimentos , Humanos , Mamografia , Pessoa de Meia-Idade , Mutação , Guias de Prática Clínica como Assunto , Valor Preditivo dos Testes , Estudos Prospectivos , Fatores de Risco , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Whether breast cancer surgeons are adequately trained, skilled, and experienced to provide breast cancer genetic assessment, testing, and counseling came under debate in September 2013 when a major third-party payer excluded nongenetics specialists from ordering such testing. A literature search having failed to uncover any study on breast surgeons' skill and practice in this area, the American Society of Breast Surgeons (ASBrS) surveyed its members on their experience with the recognized crucial components of such testing. METHODS: In late 2013, ASBrS e-mailed a link to an online questionnaire to its U.S. members (n = 2,603) requesting a self-assessment of skills and experience in genetic assessment, testing, interpretation, and counseling. After approximately 6 weeks, the results were collated and evaluated. RESULTS: By January 2, 2014, 907 responses (34.84 %) had arrived from breast surgeons nationwide working in academic settings (20 %), solo or small group private practice (39 %), large multispecialty groups (18 %), and other settings. More than half said they performed 3-generation pedigrees, ordered genetic testing, and provided pre- and posttest counseling. Most noted that they would welcome continuing educational support in genetics. CONCLUSIONS: Currently the majority of breast surgeons provide genetic counseling and testing services to their patients. They report practices that meet or exceed recognized guidelines, including the necessary elements and processes for best practices in breast cancer genetics test counseling. Because breast cancer genetic testing is grossly underutilized relative to the size of the U.S. BRCA mutation carrier population, these appropriate services should not be restricted but rather supported and expanded.