MECANISMOS MOLECULARES NO HIRSUTISMO: EXPRESSÃO GÊNICA DE ENZIMAS DO METABOLISMO ANDROGÊNICO NO FOLÍCULO PILOSO E ASSOCIAÇÃO COM O DIAGNÓSTICO ETIOLÓGICO

Androgens are the main hormonal regulators of human hair growth and they are related to clinical conditions such as hirsutism. The aim of this study was to analyze the gene expression of androgen receptor (AR), type 1 and type 2 5a-reductase isoenzymes (5a R1 and 2) and type 2 17b hydroxysteroid dehydrogenase (17b-HSD 2) in plucked scalp hairs from hirsute patients and normal subjects. We studied 33 women with hirsutism [20 with polycystic ovary syndrome (PCOS), 13 with idiopathic hirsutism (IH)]; 15 control women; and 10 control men. Hirsutism was assessed by a modified Ferriman-Gallwey method. Hormonal status was assessed between days 2 and 10 of the menstrual cycle or on any day when the patients were amenorrheic. AR and enzymes mRNA levels were estimated by reverse transcriptionpolymerase chain reaction (RT-PCR). AR expression was similar in all groups. The gene expression of 5a R2 was not detected in any hair samples analyzed in this study. No differences were found on 5a R1 mRNA levels between men and normal women (0.78 ± 0.05 vs. 0.74 ± 0.06, respectively). 5a R1 gene expression in the plucked hair cells from scalp of normal women (0.85 ± 0.04), PCOS (0.78 ± 0.05) and IH (0.80 ± 0.06) was also similar. 17b-HSD2 gene expression in hirsute patients was lower (2.2±0.13 and 2.0±0.15, for PCOS and IH, respectively) than in normal women (3.1±0.17, p<0.05), and similar to men (1.8±0.22). In conclusion, these results indicate that there are no changes on 5a R1 gene expression in the plucked hair cells from scalp, related to gender or hirsutism. The lower expression of 17b-HSD2 mRNA in scalp hairs of hirsute patients suggests androgen metabolism disturbances with predominance of more potent androgens, as occurs in men.Androgens are the main hormonal regulators of human hair growth and they are related to clinical conditions such as hirsutism. The aim of this study was to analyze the gene expression of androgen receptor (AR), type 1 and type 2 5a-reductase isoenzymes (5a R1 and 2) and type 2 17b hydroxysteroid dehydrogenase (17b-HSD 2) in plucked scalp hairs from hirsute patients and normal subjects. We studied 33 women with hirsutism [20 with polycystic ovary syndrome (PCOS), 13 with idiopathic hirsutism (IH)]; 15 control women; and 10 control men. Hirsutism was assessed by a modified Ferriman-Gallwey method. Hormonal status was assessed between days 2 and 10 of the menstrual cycle or on any day when the patients were amenorrheic. AR and enzymes mRNA levels were estimated by reverse transcriptionpolymerase chain reaction (RT-PCR). AR expression was similar in all groups. The gene expression of 5a R2 was not detected in any hair samples analyzed in this study. No differences were found on 5a R1 mRNA levels between men and normal women (0.78 ± 0.05 vs. 0.74 ± 0.06, respectively). 5a R1 gene expression in the plucked hair cells from scalp of normal women (0.85 ± 0.04), PCOS (0.78 ± 0.05) and IH (0.80 ± 0.06) was also similar. 17b-HSD2 gene expression in hirsute patients was lower (2.2±0.13 and 2.0±0.15, for PCOS and IH, respectively) than in normal women (3.1±0.17, p<0.05), and similar to men (1.8±0.22). In conclusion, these results indicate that there are no changes on 5a R1 gene expression in the plucked hair cells from scalp, related to gender or hirsutism. The lower expression of 17b-HSD2 mRNA in scalp hairs of hirsute patients suggests androgen metabolism disturbances with predominance of more potent androgens, as occurs in men

Developmental potential of in vitro or in vivo matured oocytes

Summary This study compared the embryological features of mature and immature oocytes (different stages) collected from stimulated cycles of in vitro fertilization (IVF). Immature oocytes were identified, classified as PI (prophase I -germinal vesicle, GV) or MI (metaphase I), were matured in vitro and fertilized using the intra-cytoplasmic sperm injection (ICSI) technique. Fertilization potential, cleavage, and subsequent transfer/cryopreservation of the embryos derived from these in vitro matured oocytes were compared with those of in vivo matured oocytes (collected at the MII stage). The characteristics of embryos derived from gametes recovered in the MI and MII stages were similar. The fertilization rate of immature oocytes recovered in PI was significantly lower than that of MII oocytes (P = 0.031), and the cleavage rate of the PI group was also lower than that of the MI (P = 0.004) and MII (P < 0.001) groups. In vitro maturation of MI oocytes is a suitable alternative when immature oocytes are recovered, as their characteristics and development are similar to those of in vivo matured oocytes. Optimization of outcomes for PI oocytes will require development of techniques that can distinguish which of these gametes will mature and fertilize

FMR1 expression in human granulosa cells increases with exon 1 CGG repeat length depending on ovarian reserve

Background: Fragile-X-Mental-Retardation-1- (FMR1)-gene is supposed to be a key gene for ovarian reserve and folliculogenesis. It contains in its 5’-UTR a triplet-base-repeat (CGG), that varies between 26 and 34 in general population. CGG-repeat-lengths with 55–200 repeats (pre-mutation = PM) show instable heredity with a tendency to increase and are associated with premature-ovarian-insufficiency or failure (POI/POF) in about 20%. FMR1-mRNA-expression in leucocytes and granulosa cells (GCs) increases with CGG-repeat-length in PM-carriers, but variable FMR1-expression profiles were also described in women with POI without PM-FMR1 repeat-length. Additionally, associations between low numbers of retrieved oocytes and elevated FMR1-expression levels have been shown in GCs of females with mid-range PM-CGG-repeats without POI. Effects of FMR1-repeat-lengths-deviations (n < 26 or n > 34) below the PM range (n < 55) on ovarian reserve and response to ovarian stimulation remain controversial. Methods: We enrolled 229 women undergoing controlled ovarian hyperstimulation for IVF/ICSI-treatment and devided them in three ovarian-response-subgroups: Poor responder (POR) after Bologna Criteria, polycystic ovary syndrome (PCO) after Rotterdam Criteria, or normal responder (NOR, control group). Subjects were subdivided into six genotypes according to their be-allelic CGG-repeat length. FMR1-CGG-repeat-length was determined using ALF-express-DNA-sequencer or ABI 3100/3130 × 1-sequencer. mRNA was extracted from GCs after follicular aspiration and quantitative FMR1-expression was determined using specific TaqMan-Assay and applying the ΔΔCT method. Kruskall-Wallis-Test or ANOVA were used for simple comparison between ovarian reserve (NOR, POR or PCO) and CGG-subgroups or cohort demographic data. All statistical analysis were performed with SPSS and statistical significance was set at p ≤ 0.05. Results: A statistically significant increase in FMR1-mRNA-expression-levels was detected in GCs of PORs with heterozygous normal/low-CGG-repeat-length compared with other genotypes (p = 0.044). Conclusion: Female ovarian response may be negatively affected by low CGG-alleles during stimulation. In addition, due to a low-allele-effect, folliculogenesis may be impaired already prior to stimulation leading to diminished ovarian reserve and poor ovarian response. A better understanding of FMR1 expression-regulation in GCs may help to elucidate pathomechanisms of folliculogenesis disorders and to develop risk-adjusted treatments for IVF/ICSI-therapy. Herewith FMR1-genotyping potentially provides a better estimatation of treatment outcome and allows the optimal adaptation of stimulation protocols in future

Genome-wide interrogation of structural variation reveals novel African-specific prostate cancer oncogenic drivers

ADDITIONAL FILE 1: FIGURE S1. Concordant SV call generation from Manta and GRIDSS. FIGURE S2. Summary of SVs in each type, compared to other studies. FIGURE S3. CIRCOS plot of hyper-SV mutated tumours. FIGURE S4. The spread of SV breakpoints and samples in each 1 Mbp genomic bin. FIGURE S5. TMPRSS2-ERG fusion with interstitial region retention. TABLE S1. Clinical and pathological characteristics of 180 prostate cancer patients included in this study. TABLE S2. Biallelic assessment of CDK12 in hyper-duplicated samples. TABLE S3. Biallelic assessment of BRCA2 in hyper-deleted samples.ADDITIONAL FILE 2: TABLE S4. Summary of gene fusions identified from SVs. ADDITIONAL FILE 3: TABLE S5. SV calls resulting in gene fusions.DATA AND MATERIALS AVAILABILITY : The datasets analysed in this study were obtained and accessible through Jaratlerdsiri et al [6], with sequence data deposited in the European GenomePhenome Archive (EGA; https://ega-archive.org) under overarching accession EGAS00001006425 and including the Southern African Prostate Cancer Study (SAPCS) Dataset (EGAD00001009067) and Garvan/St Vincent’s Prostate Cancer Database (EGAD00001009066). The computational code used to analyse SV subtypes, SV hotspots and gene fusions is available on GitHub [68].BACKGROUND : African ancestry is a significant risk factor for advanced prostate cancer (PCa). Mortality rates in sub- Saharan Africa are 2.5-fold greater than global averages. However, the region has largely been excluded from the benefits of whole genome interrogation studies. Additionally, while structural variation (SV) is highly prevalent, PCa genomic studies are still biased towards small variant interrogation. METHODS : Using whole genome sequencing and best practice workflows, we performed a comprehensive analysis of SVs for 180 (predominantly Gleason score ≥ 8) prostate tumours derived from 115 African, 61 European and four ancestrally admixed patients. We investigated the landscape and relationship of somatic SVs in driving ethnic disparity (African versus European), with a focus on African men from southern Africa. RESULTS : Duplication events showed the greatest ethnic disparity, with a 1.6- (relative frequency) to 2.5-fold (count) increase in African-derived tumours. Furthermore, we found duplication events to be associated with CDK12 inactivation and MYC copy number gain, and deletion events associated with SPOP mutation. Overall, African-derived tumours were 2-fold more likely to present with a hyper-SV subtype. In addition to hyper-duplication and deletion subtypes, we describe a new hyper-translocation subtype. While we confirm a lower TMPRSS2-ERG fusion-positive rate in tumours from African cases (10% versus 33%), novel African-specific PCa ETS family member and TMPRSS2 fusion partners were identified, including LINC01525, FBXO7, GTF3C2, NTNG1 and YPEL5. Notably, we found 74 somatic SV hotspots impacting 18 new candidate driver genes, with CADM2, LSAMP, PTPRD, PDE4D and PACRG having therapeutic implications for African patients. CONCLUSIONS : In this first African-inclusive SV study for high-risk PCa, we demonstrate the power of SV interrogation for the identification of novel subtypes, oncogenic drivers and therapeutic targets. Identifying a novel spectrum of SVs in tumours derived from African patients provides a mechanism that may contribute, at least in part, to the observed ethnic disparity in advanced PCa presentation in men of African ancestry.The Medical Health and Medical Research Council (NHMRC) of Australia, University of Sydney Bridging Grant, the USA. Department of Defense (DoD) Prostate Cancer Research Program (PCRP) Idea Development.https://genomemedicine.biomedcentral.comam2023School of Health Systems and Public Health (SHSPH

African-specific molecular taxonomy of prostate cancer

Data availability DNA-sequencing data have been deposited at the European Genome- Phenome Archive (EGA) under overarching accession EGAS00001006425 and including the Southern African Prostate Cancer Study (SAPCS) Dataset (EGAD00001009067 and Garvan/St Vincent’s Prostate Cancer Database EGAD00001009066). Academic researchers meeting the data-access policy criteria may apply for data access through the respective data access committees. CPGEA data are available through http://www.cpgea.com. PCAWG data are available at ICGC Data Portal (https://dcc.icgc.org/releases/PCAWG).Prostate cancer is characterized by considerable geo-ethnic disparity. African ancestry is a significant risk factor, with mortality rates across sub-Saharan Africa of 2.7-fold higher than global averages. The contributing genetic and non-genetic factors, and associated mutational processes, are unknown. Here, through whole-genome sequencing of treatment-naive prostate cancer samples from 183 ancestrally (African versus European) and globally distinct patients, we generate a large cancer genomics resource for sub-Saharan Africa, identifying around 2 million somatic variants. Significant African-ancestry-specific findings include an elevated tumour mutational burden, increased percentage of genome alteration, a greater number of predicted damaging mutations and a higher total of mutational signatures, and the driver genes NCOA2, STK19, DDX11L1, PCAT1 and SETBP1. Examining all somatic mutational types, we describe a molecular taxonomy for prostate cancer differentiated by ancestry and defined as global mutational subtypes (GMS). By further including Chinese Asian data, we confirm that GMS-B (copy-number gain) and GMS-D (mutationally noisy) are specific to African populations, GMS-A (mutationally quiet) is universal (all ethnicities) and the African–European-restricted subtype GMS-C (copy-number losses) predicts poor clinical outcomes. In addition to the clinical benefit of including individuals of African ancestry, our GMS subtypes reveal different evolutionary trajectories and mutational processes suggesting that both common genetic and environmental factors contribute to the disparity between ethnicities. Analogous to gene–environment interaction—defined here as a different effect of an environmental surrounding in people with different ancestries or vice versa—we anticipate that GMS subtypes act as a proxy for intrinsic and extrinsic mutational processes in cancers, promoting global inclusion in landmark studies.The National Health and Medical Research Council (NHMRC) of Australia, NHMRC Ideas Grants, University of Sydney Bridging Grant, the US Department of Defense (DoD) Prostate Cancer Research Program (PCRP) Idea Development Award TARGET Africa.http://www.nature.com/natuream2023School of Health Systems and Public Health (SHSPH

MECANISMOS MOLECULARES NO HIRSUTISMO: EXPRESSÃO GÊNICA DE ENZIMAS DO METABOLISMO ANDROGÊNICO NO FOLÍCULO PILOSO E ASSOCIAÇÃO COM O DIAGNÓSTICO ETIOLÓGICO

Androgens are the main hormonal regulators of human hair growth and they are related to clinical conditions such as hirsutism. The aim of this study was to analyze the gene expression of androgen receptor (AR), type 1 and type 2 5a-reductase isoenzymes (5a R1 and 2) and type 2 17b hydroxysteroid dehydrogenase (17b-HSD 2) in plucked scalp hairs from hirsute patients and normal subjects. We studied 33 women with hirsutism [20 with polycystic ovary syndrome (PCOS), 13 with idiopathic hirsutism (IH)]; 15 control women; and 10 control men. Hirsutism was assessed by a modified Ferriman-Gallwey method. Hormonal status was assessed between days 2 and 10 of the menstrual cycle or on any day when the patients were amenorrheic. AR and enzymes mRNA levels were estimated by reverse transcriptionpolymerase chain reaction (RT-PCR). AR expression was similar in all groups. The gene expression of 5a R2 was not detected in any hair samples analyzed in this study. No differences were found on 5a R1 mRNA levels between men and normal women (0.78 ± 0.05 vs. 0.74 ± 0.06, respectively). 5a R1 gene expression in the plucked hair cells from scalp of normal women (0.85 ± 0.04), PCOS (0.78 ± 0.05) and IH (0.80 ± 0.06) was also similar. 17b-HSD2 gene expression in hirsute patients was lower (2.2±0.13 and 2.0±0.15, for PCOS and IH, respectively) than in normal women (3.1±0.17, p<0.05), and similar to men (1.8±0.22). In conclusion, these results indicate that there are no changes on 5a R1 gene expression in the plucked hair cells from scalp, related to gender or hirsutism. The lower expression of 17b-HSD2 mRNA in scalp hairs of hirsute patients suggests androgen metabolism disturbances with predominance of more potent androgens, as occurs in men.Androgens are the main hormonal regulators of human hair growth and they are related to clinical conditions such as hirsutism. The aim of this study was to analyze the gene expression of androgen receptor (AR), type 1 and type 2 5a-reductase isoenzymes (5a R1 and 2) and type 2 17b hydroxysteroid dehydrogenase (17b-HSD 2) in plucked scalp hairs from hirsute patients and normal subjects. We studied 33 women with hirsutism [20 with polycystic ovary syndrome (PCOS), 13 with idiopathic hirsutism (IH)]; 15 control women; and 10 control men. Hirsutism was assessed by a modified Ferriman-Gallwey method. Hormonal status was assessed between days 2 and 10 of the menstrual cycle or on any day when the patients were amenorrheic. AR and enzymes mRNA levels were estimated by reverse transcriptionpolymerase chain reaction (RT-PCR). AR expression was similar in all groups. The gene expression of 5a R2 was not detected in any hair samples analyzed in this study. No differences were found on 5a R1 mRNA levels between men and normal women (0.78 ± 0.05 vs. 0.74 ± 0.06, respectively). 5a R1 gene expression in the plucked hair cells from scalp of normal women (0.85 ± 0.04), PCOS (0.78 ± 0.05) and IH (0.80 ± 0.06) was also similar. 17b-HSD2 gene expression in hirsute patients was lower (2.2±0.13 and 2.0±0.15, for PCOS and IH, respectively) than in normal women (3.1±0.17, p<0.05), and similar to men (1.8±0.22). In conclusion, these results indicate that there are no changes on 5a R1 gene expression in the plucked hair cells from scalp, related to gender or hirsutism. The lower expression of 17b-HSD2 mRNA in scalp hairs of hirsute patients suggests androgen metabolism disturbances with predominance of more potent androgens, as occurs in men

Expressão dos protooncogenes c-fos, c-myc e c-jun em miométrio normal e mioma humanos

OBJETIVO: Comparar a expressão gênica (mRNA) e protéica dos protooncogenes c-fos, c-myc e c-jun em miométrio normal e mioma humanos. MÉTODOS: Foi realizado um estudo do tipo caso-controle. O material foi coletado de 12 pacientes submetidas a histerectomia no Hospital de Clínicas de Porto Alegre. A expressão do mRNA específico para c-myc, c-fos, c-jun e beta-microglobulina foi avaliada pela técnica de RT-PCR, utilizando primers específicos para cada gene. A expressão protéica destes protooncogenes foi avaliada através de Western blot com anticorpos específicos. RESULTADOS: Não houve diferença significativa para expressão gênica desses protooncogenes entre miométrio normal e mioma (c-myc: 0,87 ± 0,08 vs 0,87 ± 0,08, p = 0,952; c-fos: 1,10 ± 0,17 vs 1,01 ± 0,11, p = 0,21; c-jun: 1,03 ± 0,12 vs 0,96 ± 0,09, p = 0,168, respectivamente). Não houve diferença significativa para expressão protéica desses protooncogenes entre miométrio normal e mioma (c-myc: 1,36 ± 0,48 vs 1,53 ± 0,29, p = 0,569; c-fos: 8,85 ± 5,5 vs 6,56 ± 4,22, p = 0,434; e c-jun: 6,47 ± 3,04 vs 5,42 ± 2,03, p = 0,266, respectivamente). CONCLUSÃO: A expressão gênica (transcrição) e a expressão protéica (tradução) dos protooncogenes c-myc, c-fos e c-jun em mioma e miométrio normal são semelhantes