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Cargo small non-coding RNAs of extracellular vesicles isolated from uterine fluid associate with endometrial receptivity and implantation success

      Objective

      To optimize a method of isolating extracellular vesicles (EVs) from uterine fluid and to characterize small non-coding RNAs (sncRNAs) from the EVs, with the goal of identifying novel receptivity-associated biomarkers.

      Design

      Longitudinal study comparing sncRNA expression profiles from endometrial EVs.

      Setting

      University-affiliated, hospital-based fertility clinic.

      Patient(s)

      Healthy volunteers with no history of infertility (Group A) and women receiving controlled ovarian stimulation (COS)–in vitro fertilization treatment (Group B).

      Interventions(s)

      In Group A, EVs were isolated from uterine fluid obtained on luteinizing hormone (LH)+2 and LH+7 in one natural menstrual cycle. In Group B, EVs were isolated from uterine fluid obtained on human chorionic gonadotropin (hCG)+2 and hCG+7 in one COS cycle. RNAs extracted from EVs were profiled using next-generation sequencing.

      Main Outcome Measure(s)

      Differential EV-sncRNAs between LH+2 and LH+7 (Group A), between hCG+2 and hCG+7 (Group B), and between pregnant and nonpregnant in vitro fertilization cycles (Group B).

      Result(s)

      Ultracentrifugation was validated as the most efficient method to isolate EVs from uterine fluid. We identified 12 endometrial EV-sncRNAs (11 microRNAs and 1 piwi-interacting RNA) as receptivity-associated transcripts conserved in both natural and COS cycles. These sncRNAs were associated strongly with biological functions related to immune response, extracellular matrix, and cell junction. Within COS cycles, we also identified a group of EV-sncRNAs that exhibited differential expression in patients who conceived versus those who did not, with hsa-miR-362-3p most robustly overexpressed in the nonpregnant patients.

      Conclusion(s)

      This study is the first to profile comprehensively sncRNAs in endometrial EVs from uterine fluid and identify sncRNA biomarkers of endometrial receptivity and implantation success.
      Los pequeños ARN cargo no codificantes de vesículas extracelulares aisladas del líquido uterino se asocian con la receptividad endometrial y el éxito de la implantación.

      Objetivo

      Optimizar un método para aislar las vesículas extracelulares (VE) del fluido uterino y caracterizar los ARN pequeños no codificantes (sncRNA) de los VE, con el objetivo de identificar nuevos biomarcadores asociados a la receptividad.

      Diseño

      Estudio longitudinal que compara los perfiles de expresión de sncRNA de EV endometriales.

      Entorno

      Clínica de fertilidad hospitalaria afiliada a una universidad.

      Paciente (s)

      Voluntarias sanas sin antecedentes de infertilidad (Grupo A) y mujeres recibiendo tratamiento de fertilización in vitro con estimulación ovárica controlada (COS) - (Grupo B).

      Intervención (es)

      En el Grupo A, los VE se aislaron del líquido uterino obtenido con la hormona luteinizante (LH) +2 y LH +7 en un ciclo menstrual natural. En el Grupo B, los EV se aislaron del líquido uterino obtenido con gonadotropina coriónica humana (hCG) +2 y hCG +7 en un ciclo COS. Los ARN extraídos de las VEs se perfilaron mediante secuenciación de próxima generación.

      Principales medidas de resultado

      EV-sncRNA diferenciales entre LH +2 y LH +7 (Grupo A), entre hCG +2 y hCG +7 (Grupo B), y entre embarazadas y no embarazadas (Grupo B) de ciclos de fertilización in vitro.

      Resultado (s)

      La ultracentrifugación se validó como el método más eficaz para aislar las EVs del líquido uterino. Identificamos 12 EV-sncRNAs de endometrio (11 microRNAs y 1 RNA que interactúa con piwi) como transcripciones asociadas a la receptividad conservadas en ciclos tanto naturales como de COS. Estos sncRNAs se asociaron fuertemente con funciones biológicas relacionadas con la respuesta inmune, la matriz extracelular y la unión celular. Dentro de los ciclos de COS, también identificamos un grupo de EV-sncRNA que exhibieron expresión diferencial en pacientes que concibieron versus aquellas que no, con hsa-miR-362-3p sobreexpresado de manera más robusta en las pacientes no embarazadas.

      Conclusión (es)

      Este estudio es el primero en perfilar de forma exhaustiva los sncARNs en los EV endometriales del líquido uterino e identificar los biomarcadores sncRNA de receptividad endometrial y éxito de la implantación.

      Palabras clave

      Líquido uterino, vesículas extracelulares, pequeños ARN no codificantes, receptividad, implantación

      Key Words

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