Mild hyperandrogenemia in presence/absence of a high-fat, Western-style diet alters secretory phase endometrial transcriptome in nonhuman primates

Abstract

Objective: To identify novel transcriptomic changes to eutopic endometrium by exposure to chronic mild hypernadrogenemia (testosterone [T]) with/without exposure to an obesogenic Western-style diet (WSD). Design: Two-by-two factorial arrangement of treatments. Setting: National primate research center. Animal(s): Rhesus macaque females were chronically exposed to T and/or consumed a WSD from menarche through adulthood. After 4.5 years of treatment, Tru-Cut endometrial biopsies were obtained at the midsecretory phase (n ¼ 6-4/group), and paired-end sequencing of RNA was performed. Several females in the T, WSD, and TþWSD cohorts developed endometriosis within 6 months of biopsy; a separate analysis was performed contrasting diagnosis of endometriosis stage 0-2 versus stages 3 and 4 (American Society for Reproductive Medicine revised criteria). Intervention(s): Chronic exposure to mild elevation of T ($five-fold elevation) and/or WSD from menarche until adulthood. Main Outcome Measure(s): Limma voom empirical Bayes pipeline was performed to detect differentially expressed RNAs (DEs) significantly impacted by treatments and endometriosis severity. Differentially expressed RNAs were then interrogated by Ingenuity Pathway Analyses and Protein Analysis through Evolutionary Relationships. Result(s): Total DEs included C versus T, 469; C versus WSD, 525; C versus TþWSD, 549; and T versus TþWSD, 1,505. The majority of DEs mapped to the ontology pathways: heterotrimeric G-protein signaling pathways Gi alpha and Gs alpha (C vs. T), WNT signaling (C vs. WSD and T vs. TþWSD), and Huntington disease (C vs. TþWSD). A total of 2,171 DEs from eutopic endometrium were altered by the presence of stage 3 and 4 endometriosis lesions. Conclusion(s):The present global transcriptomic analyses demonstrate that the greatest magnitude of changes occurred in contrasts of C and T versus TþWSD, adding to the evidence that these two insults have a synergistic effect on female physiology. These data also support the concept that prior alterations to the function of eutopic endometrium increase the risk for endometriosis. (Fertil Steril Sci Ò 2020;1:172-82. Ó2020 by American Society for Reproductive Medicine.

Utilizing Contrast-Enhanced Ultrasonography with Phosphatidylserine Microbubbles to Detect Placental Inflammation in Rhesus Macaques

“…We utilized rhesus macaques ( Macaca mulatta ) exposed to long-term treatment with either a control chow diet or an iso-caloric, fat rich, western-style diet (WSD) in the presence or absence of a subcutaneous implant filled with either cholesterol or testosterone (T) to emulate polycystic ovary syndrome (PCOS). The details of the animal treatments were previously published [ 54 , 55 , 56 , 57 , 58 ]. Briefly, the dietary and T treatments started near menarche and continued for 7 years, at which time the ultrasound procedures were performed.…”

Correction to: The combined impact of testosterone and Western-style diet on endometriosis severity and progression in rhesus macaques

“…Abdominal cavities were examined during annual laparoscopic procedures, and abdominal ultrasound was performed twice yearly by experienced staff (CVB and HS). Observations were used to stage endometriosis as previously published [16].…”

“…Staging for endometriosis was done similarly to the ASRM revised criteria for endometriosis, modified for rhesus females [16]. Briefly, minor abdominal adhesions were considered stage 1, extensive adhesions (but no endometriotic cysts) were considered stage 2, presence of chocolate cysts <1 cm were considered stage 3, and presence of multiple large >1 cm chocolate cysts surrounding reproductive tract (usually accompanied by extensive adhesions) were considered stage 4.…”

“…Laparoscopic and ultrasound images of reproductive tracts with these characteristics were published previously (Supplemental Figure 1 in ref. [16]).…”

“…Interestingly, during this multi-year study we observed a high prevalence of endometriosis, particularly in the T + WSD cohort which had a higher prevalence of metabolic dysfunction than other treatment groups. [16]. Endometrial progesterone resistance is associated with endometriosis in humans [17,18] and NHPs, including the baboon [19].…”