Paulo D'Amora

Genetic polymorphisms of cytochrome P450c17α (CYP17) and progesterone receptor genes (PROGINS) in the assessment of endometriosis risk

We designed the present study in order to evaluate the eventual role of polymorphisms in the genes encoding cytochrome P450c17α (CYP17) and the progesterone receptor (PROGINS) as risk factors for endometriosis development. Eligible cases consisted of 121 women with surgically confirmed endometriosis who underwent treatment in a hospital in São Paulo, Brazil during the period from September 2003 to September 2005. The 281 controls were participants with normal gynecological as well as pelvic ultrasound evaluation, who did not have any gynecological conditions during their reproductive lives such as pelvic pain and/or dyspareunia nor infertility history. Genomic DNA was obtained from buccal cells and processed for DNA extraction using the GFX DNA extraction kit (GE Healthcare). The CYP17 (−34T→C) polymerase chain reaction–restriction fragment length polymorphism assay has been described previously, as has the progesterone receptor polymorphism (PROGINS) detection assay. PROGINS heterozygosis genotype frequencies were shown to be statistically higher in endometriosis cases compared with controls. On the other hand, differences in the CYP17 polymorphism (−34T → C) frequencies were not even close to significance (p = 0.278) according to our findings.

Polymorphisms in exons 1B and 1C of the type I interleukin-1 receptor gene in patients with endometriosis.

Problem  To study possible correlation between the prevalence of polymorphisms in the type I interleukin‐1 receptor gene and pelvic endometriosis.

Disrupted Cell Cycle Control in Cultured Endometrial Cells from Patients with Endometriosis Harboring the Progesterone Receptor Polymorphism PROGINS

Presently, little is understood about how endometriosis is established or maintained, or how genetic factors can predispose women to the disease. Because of the crucial role that the progesterone receptor polymorphism PROGINS plays in predisposing women to the development of endometriosis, we hypothesized that this variant may influence critical steps during endometrial cell metabolism that are involved in the pathogenesis of endometriosis. Eutopic endometria were collected from three sources: women with endometriosis who had a single PROGINS allele (from the progesterone receptor gene); women with endometriosis who had the wild-type progesterone receptor allele; and women without endometriosis who had the wild-type allele. Cells prepared from the eutopic endometria of these women were stimulated with both estradiol and progesterone, and then examined for cell proliferation, viability, and apoptosis. The cells from women with endometriosis that carried the PROGINS allele demonstrated increased proliferation, greater viability, and decreased apoptosis following progesterone treatment. In general, these parameters were very different as compared with those of women with endometriosis but without the PROGINS allele and women in the control group. This result indicates there is a reduced level of progesterone responsiveness in women who carry the PROGINS polymorphism. Because progesterone responsiveness is known to be an important characteristic of women with endometriosis, these data support the contention that the PROGINS polymorphism enhances the endometriosis phenotype.

Intron 1 and exon 1 alpha estrogen receptor gene polymorphisms in women with endometriosis

OBJECTIVE To evaluate the association of intron 1 and exon 1 polymorphisms in the estrogen receptor alpha gene (ER-alpha) with endometriosis in women. DESIGN Association study. SETTING Endometriosis Unit, Federal University of São Paulo. PATIENT(S) The control group consisted of volunteers older than 45 years who had no evidence of endometriosis antecedents. Two groups with the disease were evaluated: the first group had stage I or II endometriosis and the second group stage III or IV. INTERVENTION(S) Polymerase chain reaction (PCR) followed by digestion with HaeIII and MspI endonucleases (RFLP) were applied to detect intron 1 and exon 1 polymorphisms, respectively, in a total of 125 controls and 105 affected women. MAIN OUTCOME MEASURE(S) Frequency and distribution of HaeIII and MspI polymorphisms in ER-alpha. RESULT(S) No significant differences in the frequency of polymorphisms either in intron 1 or exon 1 of ER-alpha were found when endometriosis patients were compared with control subjects. Furthermore, the frequency of ER-alpha polymorphisms within the two different groups of patients with disease was statistically similar. The odds ratio between presence of intron 1 single-nucleotide polymorphisms (SNP) and endometriosis was 0.904, and the odds ratio between exon 1 SNP and endometriosis was 0.976. CONCLUSION(S) The evaluated polymorphisms were not associated with endometriosis.

TP53 gene polymorphisms at codons 11, 72, and 248 and association with endometriosis in a Brazilian population

We evaluated the association between TP53 gene polymorphisms and endometriosis in Brazilian women. Genomic DNA was extracted from swabs of buccal cells collected from hospital patients. TP53 gene polymorphisms were investigated at three codons: TP53 11 Glu/Gln or Lys (GAG->CAG or AAG), TP53 72 Arg/Pro (CCG->CCC), and TP53 248 Arg/Thr (CGG->TCG) using the polymerase chain reaction-restriction fragment length polymorphism method. TP53 11 presented the following genotypic distribution: the control group was 98.28% homozygous wild-type (Glu) and 1.72% homozygous variant (Gln/Lys), and the heterozygous genotype was not identified. The genotypic distribution in the endometriosis group was 96% homozygous wild-type (Glu) and 4% heterozygous (Glu-Gln/Lys); the homozygous variant genotype was not identified (P = 0.02). TP53 72 showed the following genotypic distribution: the control group was 29.75% homozygous wild-type (Arg), 47.11% heterozygous (Arg-Pro), and 23.14% homozygous variant (Pro). The genotypic distribution in the endometriosis group was 16.15% homozygous wild-type (Arg), 51.54% heterozygous (Arg-Pro), and 32.31% homozygous variant (Pro) (odds ratio = 2.26; 95% confidence interval = 1.19-4.03; P = 0.02). Only one patient had the homozygous TP53 248 genotype (Arg-Trp/Gln); all other patients were homozygous wild-type in both the control and endometriosis groups (P = 0.51; NS). We found that TP53 72 polymorphism may be associated with susceptibility to endometriosis; the presence of at least 1 polymorphic allele increased the chance of disease development by 2.26-fold. Hence, this genetic variant is a potential candidate marker for endometriosis.

Polimorfismo do gene do receptor de progesterona (PROGINS) em mulheres com endometriose pélvica

OBJECTIVE: the aim of the present study was to verify whether there is a correlation between the prevalence of the polymorphism in the progesterone receptor gene named PROGINS and pelvic endometriosis at different stages. METHODS: a case-control study carried out from November 2003 to May 2004. The genotypes of 104 women were analyzed 66 women had had surgically confirmed endometriosis (26 women at stages I-II and 40 at stages III-IV), and 38 were healthy women. The 306-base pair Alu insertion polymorphism in the intron G of the progesterone receptor gene was detected by polymerase chain reaction and analyzed on 2% agarose gel stained with ethidium bromide. ANOVA analysis was performed in order to make comparisons between among the studied groups. RESULTS: the groups of women with endometriosis stages I-II (EndoI group) and stages III-IV (EndoII group) showed statistically significant increased incidence of PROGINS polymorphic allele as compared with the control group: 27% in the EndoI group, 38% in EndoII and 18% in the control group (p < 0.001. In the analyses, a high frequency of the PROGINS insertion was observed in women with endometriosis as compared to healthy women, disregarding the clinical stage of the disease (p = 0.0385). CONCLUSION: there is a significant statistical association between the PROGINS polymorphism and pelvic endometriosis.

Inborn-like errors of metabolism are determinants of breast cancer risk, clinical response and survival: a study of human biochemical individuality

Breast cancer remains a leading cause of morbidity and mortality worldwide yet methods for early detection remain elusive. We describe the discovery and validation of biochemical signatures measured by mass spectrometry, performed upon blood samples from patients and controls that accurately identify (>95%) the presence of clinical breast cancer. Targeted quantitative MS/MS conducted upon 1225 individuals, including patients with breast and other cancers, normal controls as well as individuals with a variety of metabolic disorders provide a biochemical phenotype that accurately identifies the presence of breast cancer and predicts response and survival following the administration of neoadjuvant chemotherapy. The metabolic changes identified are consistent with inborn-like errors of metabolism and define a continuum from normal controls to elevated risk to invasive breast cancer. Similar results were observed in other adenocarcinomas but were not found in squamous cell cancers or hematologic neoplasms. The findings describe a new early detection platform for breast cancer and support a role for pre-existing, inborn-like errors of metabolism in the process of breast carcinogenesis that may also extend to other glandular malignancies. Statement of Significance: Findings provide a powerful tool for early detection and the assessment of prognosis in breast cancer and define a novel concept of breast carcinogenesis that characterizes malignant transformation as the clinical manifestation of underlying metabolic insufficiencies.

Abstract TMEM-021: CORRELATION BETWEEN DRUG SENSITIVITY AND METABOLOMIC SIGNATURES WITH OBJECTIVE RESPONSE (OR), TIME TO PROGRESSION (TTP), AND OVERALL SURVIVAL (OS) IN ADVANCED EPITHELIAL OVARIAN CANCER (EOC): A PHASE II STUDY

INTRODUCTION: Personalized oncology has advanced through the application of genomic, transcriptomics & proteomic platforms. BCR-abl; EGFr & ALK have provided drug-able targets & companion diagnostics in several diseases, yet many human transforming events are polygenic & incompletely understood at a genetic level. The recognition that oncogenesis reflects changes in both the cell and its micro-environment has renewed interest in whole cell experimental models that capture native-state cell-cell, -stroma & -vascular signaling. Phenotypic platforms like the Ex vivo Analysis of Programmed Cell Death (EVA-PCD) and Metabolomic Analyses have been shown to correlate significantly with response, time to progression & survival in cancer patients. PURPOSE: To correlate drug response profiles measured by EVA-PCD with Metabolomic Profiles measured by Mass Spectrometry, in patients with newly diagnosed, advanced epithelial ovarian cancer (EOC). Procedures: After signing informed consent, patients with advanced EOC undergoing cytoreductive surgery submit tumor tissue for EVA/PCD and blood samples for Mass Spectrometry. EVA/PCD analyses using morphologic (delayed loss of membrane integrity) and metabolic (ATP-content, mitochondrial metabolism) endpoints are used to generate dose-response curves, interpolated to provide LC50 values. The LC50 values for Cisplatin and the principal platinum-based combinations are used to define drug sensitive/drug resistant profiles for correlation with plasma-sample lipidomic analytes measured by Mass Spectrometry. Metabolite profiles from 21 participants are under analysis for more than 180 different circulating components including: amino acids, acylcarnitines, phosphatidylcholines, sphingomyelins and hexoses simultaneously identified/quantified in a targeted approach by UPLC/Mass Spectrometry. Associations between metabolite concentrations and drug sensitivity/resistance as well as clinical response parameters in EOC patients are being analyzed using adjusted linear regression models. Pairwise relationships among metabolites are investigated and illustrated by Gaussian graphical models (GGMs). RESULTS: To date, 21EOC patients have been accrued and are under analysis with a target accrual of 50. CONCLUSIONS: The growing recognition that malignant transformation represents changes in cellular metabolism offers the opportunity to interrogate human tumor biology at the level of metabolomics. This study will be among the first to correlate EOC drug response, time to progression and survival with metabolomic profiles and may offer new insights into ovarian carcinogenesis and cellular mechanisms of drug resistance with the potential to develop new predictive & prognostic models for advanced EOC. Funding: Sao Paulo State Research Foundation (FAPESP): grants 2014/19171-2 / 2015/16921-3 (Sao Paulo, SP, Brazil) and grants from Memorial Medical Center Foundation (Long Beach, CA, USA). Citation Format: Paulo D9Amora MD, PhD; Ismael DCG Silva MD, PhD; 142 MD, PhD; Marcia Batista Salzgeber; Manoel JBC Girao MD, PhD; Robert Bristow MD; Steven Evans BS, MA; Philip J. DiSaia MD and Robert Nagourney MD. CORRELATION BETWEEN DRUG SENSITIVITY AND METABOLOMIC SIGNATURES WITH OBJECTIVE RESPONSE (OR), TIME TO PROGRESSION (TTP), AND OVERALL SURVIVAL (OS) IN ADVANCED EPITHELIAL OVARIAN CANCER (EOC): A PHASE II STUDY [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr TMEM-021.

Abstract 266: Angiotensin I-7 modulates transcriptional (de)activation of multiple members of steroid hormone receptor superfamily: Possible mechanisms underlying inhibited proliferation in (T47D) human breast cancer cells

Introduction: Renin Angiotensin System (RAS) is correlated to breast cancer and to other types of cancer. Angiotensin I-7 [Ang-(I-7)] is an endogenous 7-amino acid peptide hormone of this system that has anti-proliferative properties. Interestingly, there are increasing evidences that RAS is implicated in the development of breast cancer. Objectives: We aimed to evaluate the expression of steroid hormone receptors (progesterone receptors (PR), estrogen receptors (ER), androgen receptors (AR), apoptosis and proliferation in tumoral (T47D) epithelial mammary cell line, after Ang-(I-7) treatment. Methods: Tumoral (T47D) cultured cell line was treated with angiotensin I-7 peptide and cells were harvested after 2, and 15 days, when expression analysis of steroid hormone receptors was performed by real time PCR. The same treatment was used for the apoptosis and cell cycle analyses, done using the 5HT-GUAVA flow cytometer. Results: Expression of steroid hormone receptors implicated to breast carcinogenesis such as genomic progesterone receptors (PR(A+B) and PRB), estrogen receptors (Era and ERb), androgen receptor (AR) were never investigated together in the sex hormone-sensitive human breast cancer cell T47D after angiotensin I-7 exposure. When T47D cells were treated with angiontensin I-7 peptide were all up-regulated of the control level (PRA+B (1.0-fold), PRB (2.3-fold), Era (7.8-fold), ERb (8.1-fold) and AR (1.9-fold) - p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 266. doi:1538-7445.AM2012-266