RESUMEN
PURPOSE: A symbiotic relationship between ovarian granulosa cells (GC) and the developing oocyte is critical. Genetic modulations in GC's can lead to reproductive insufficiency, highlighting the role of GC's in reproductive competence. Utilizing gene expression analyses in cumulus GC's, we attempt to enhance our understanding of mechanisms that may contribute to poor reproductive capacity in young women with diminished ovarian reserve (DOR). METHODS: We measured gremlin 1 (GREM1) gene expression in GC's from infertile women <38 years undergoing in vitro fertilization in the context of DOR. RESULTS: GREM1, a member of the differential screening-selected gene aberrative in neuroblastoma (DAN) family of genes known for its highly regulated expression pattern during folliculogenesis and a downstream effecter of oocyte-derived growth and differentiation factor 9, was down-regulated 3-fold (-3.08) in women with DOR versus control; down-regulation was confirmed by qRT-PCR (-4.02). CONCLUSION: This is the first demonstration linking differential expression of Gremlin with etiology of infertility in women.
Asunto(s)
Regulación hacia Abajo , Infertilidad Femenina/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Oocitos/crecimiento & desarrollo , Células del Cúmulo/citología , Células del Cúmulo/metabolismo , Femenino , Fertilización In Vitro , Factor 9 de Diferenciación de Crecimiento/genética , Factor 9 de Diferenciación de Crecimiento/metabolismo , Humanos , Infertilidad Femenina/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Oocitos/metabolismo , Oogénesis/genética , Ovario/citología , EmbarazoRESUMEN
OBJECTIVE: To investigate if a diagnosis of diminished ovarian reserve (DOR) is associated with a differential gene profile of ovarian granulosa cells (GCs) in infertile women undergoing in vitro fertilization (IVF). DESIGN: Prospective Cohort Study. SETTING: Academic IVF Program. PATIENTS: Infertile women <38 years were prospectively enrolled into 2 groups: normal ovarian reserve (NOR, follicle-stimulating hormone [FSH] < 10 mIU/mL, n = 4) and DOR (FSH ≥ 10.0 mIU/mL, n = 4). INTERVENTIONS: Cumulus (C) and mural (M) GCs were isolated at egg retrieval; messenger RNA was extracted and transcribed. MAIN OUTCOME MEASURE(S): Differential gene expression in cerebellar granule cells (CGCs) in the 2 groups was assessed by cDNA microarray. Microarray findings were validated by quantitative real-time polymerase chain reaction (qRTPCR) in CGCs and explored in multinucleated giant cells (MGCs). RESULTS: Of the 1256 differentially regulated genes identified in CGCs of women with DOR, the insulin-like growth factor (IGF) family was a biologically relevant gene family of a priori interest. Downregulation of IGF1 and IGF2 ligands (-3.28- and -2.54-fold, respectively), and their receptors, (-3.53- and -1.32-fold downregulation of IGF1R and IGF2R, respectively) was identified in luteinized CGCs in women with DOR compared to those with NOR. Downregulation of both IGF1 and IGF 2 ligands (-4.35- and 3.89-fold, respectively) was furthermore observed in MGCs in women with DOR compared to those with NOR; no differences in the expression of respective receptors were however observed in MGCs in the 2 groups. CONCLUSIONS: Components of the IGF gene family are downregulated in GCs of women with DOR. These findings maybe contributory to the reproductive compromise observed in women with DOR, and merit further exploration.
Asunto(s)
Regulación de la Expresión Génica , Células de la Granulosa/fisiología , Infertilidad Femenina/genética , Folículo Ovárico/fisiología , Receptores de Somatomedina/biosíntesis , Somatomedinas/biosíntesis , Adulto , Estudios de Cohortes , Femenino , Células de la Granulosa/metabolismo , Humanos , Infertilidad Femenina/metabolismo , Folículo Ovárico/citología , Folículo Ovárico/metabolismo , Estudios Prospectivos , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Receptores de Somatomedina/genética , Somatomedinas/genéticaRESUMEN
PURPOSE: Relationships between follicular fluid levels of IL-6 with ovarian response and clinical pregnancy were evaluated in IVF-ET cycles. METHODS: Follicular fluid was collected from ovarian follicles ≥ 14 mm, pooled for each patient, and IL-6 levels were assessed using ELISA (n = 68). Relationships between IL-6 levels and IVF cycle parameters were assessed using nonparametric tests, and between IL-6 levels and clinical pregnancy using multivariable logistic regression analyses. RESULTS: Significant positive correlations were observed between IL-6 with age (p = 0.035), and IL-6 with estradiol on the day of hCG (p = 0.011). On adjusted analyses, IVF cycles in patients with IL-6 levels <4.0 pg/ml (median value for the group) demonstrated an almost 4-fold increase in likelihood for clinical pregnancy (p = 0.045). CONCLUSIONS: Lower follicular fluid IL-6 levels in IVF patients are associated with increased likelihood of clinical pregnancy. We hypothesize that endometrial receptivity is a likely target for any deleterious influences of elevated IL-6 levels.
Asunto(s)
Fertilización In Vitro , Líquido Folicular/metabolismo , Interleucina-6/análisis , Adulto , Estudios Transversales , Transferencia de Embrión , Estradiol/sangre , Femenino , Humanos , Modelos Logísticos , Análisis Multivariante , Folículo Ovárico , Embarazo , Estudios RetrospectivosRESUMEN
OBJECTIVE: To determine whether 25OH-D levels in the follicular fluid (FF) of infertile women undergoing IVF demonstrate a relationship with IVF cycle parameters and outcome, hypothesizing that levels of 25OH-D in body fluids are reflective of vitamin repletion status. DESIGN: Prospective cohort study. SETTING: Academic tertiary care center. PATIENT(S): Eighty-four infertile women undergoing IVF. INTERVENTION(S): Follicular fluid from follicles>or=14 mm; serum (n = 10) and FF levels of 25OH-D. MAIN OUTCOME MEASURE(S): Clinical pregnancy (CP), defined as evidence of intrauterine gestation sac on ultrasound, following IVF; IVF cycle parameters. RESULT(S): Serum and FF levels of 25OH-D were highly correlated (r=0.94). In a predominantly Caucasian population (66%), significantly lower FF 25OH-D levels were noted in Black versus non-Black patients. Significant inverse correlations were seen between FF 25OH-D levels and body mass index (r=-0.25). Significantly higher CP and implantation rates were observed across tertiles of FF25OH-D; patients achieving CP following IVF (n=26) exhibited significantly higher FF levels of 25OH-D. Multivariable logistic regression analysis confirmed FF 25OH-D levels as an independent predictor to success of an IVF cycle; adjusting for age, body mass index, ethnicity, and number of embryos transferred, each ng/mL increase in FF 25OH-D increased the likelihood for achieving CP by 6%. CONCLUSION(S): Our findings that women with higher vitamin D level in their serum and FF are significantly more likely to achieve CP following IVF-embryo transfer are novel. A potential for benefit of vitamin D supplementation on treatment success in infertile patients undergoing IVF is suggested and merits further investigation.
Asunto(s)
Fertilización In Vitro , Líquido Folicular/química , Infertilidad Femenina/diagnóstico , Infertilidad Femenina/terapia , Vitamina D/análisis , Adulto , Estudios de Cohortes , Implantación del Embrión/fisiología , Femenino , Líquido Folicular/metabolismo , Humanos , Infertilidad Femenina/sangre , Infertilidad Femenina/metabolismo , Valor Predictivo de las Pruebas , Embarazo , Índice de Embarazo , Reproducción/fisiología , Resultado del Tratamiento , Vitamina D/sangre , Vitamina D/metabolismoRESUMEN
Declining reproductive success with aging is attributable to qualitative and quantitative deterioration in oocytes, which are nurtured by granulosa cells (GCs). This prospective study assesses whether reproductive aging is accompanied by differential gene expression in luteinized GCs from in vitro fertilization (IVF) patients. Women with nonovarian infertility etiologies were categorized as younger (< or =30, n = 3) or older (> or =40, n = 3). During oocyte retrieval, mural GCs were isolated; messenger RNA (mRNA) was extracted and transcribed for complementary DNA (cDNA) microarray analysis. Differential gene expression was confirmed by real-time polymerase chain reaction (PCR). Analysis revealed 120 genes were differentially expressed. Three genes were upregulated and 117 were downregulated (including interleukin [IL]-1beta, IL-1R2, and IL-6R) in GCs of older versus younger patients. Our data provide evidence of downregulation in IL-1 and IL-6 gene families in luteinized GCs with advancing age. Given previously recognized roles for the IL gene family in folliculogenesis and ovulation, our findings may partly explain ovulatory and luteal dysfunctions associated with reproductive aging.
Asunto(s)
Envejecimiento/genética , Regulación del Desarrollo de la Expresión Génica/genética , Células de la Granulosa/metabolismo , Interleucina-1/genética , Interleucina-6/genética , Adulto , Envejecimiento/metabolismo , Regulación hacia Abajo/genética , Femenino , Fertilización In Vitro , Expresión Génica/genética , Humanos , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia Arriba/genéticaRESUMEN
OBJECTIVE: To identify if declining ovarian reserve adversely influences granulosa cell (GC) parameters in patients undergoing IVF. DESIGN: Prospective study. SETTING: Academic IVF Center. PATIENT(S): Twenty-four women undergoing IVF. INTERVENTION(S): Mural and cumulus GCs were harvested (at time of egg retrieval). MAIN OUTCOME MEASURE(S): Influence of ovarian reserve, reflected by maximal basal FSH levels, on GC (cumulative, mural, and cumulus) counts and viability were investigated. RESULT(S): The FSH levels demonstrated an inverse correlation with cumulative and mural, but not cumulus, GC viability. Adverse influence of increasing FSH on cumulative GC viability was independent of patient's age. The GC counts were uninfluenced by ovarian reserve status. Neither GC count nor viability demonstrated any relationship with clinical pregnancy after IVF. CONCLUSION(S): We observed an adverse association between declining ovarian reserve and GC parameters. We believe these findings may partly explain the suboptimal reproductive performance in these women.