INFERTILITY TREATMENT USING HUMAN MESENCHYMAL STEM CELL DERIVED EXTRACELLULAR VESICLE IN LETROZOLE INDUCED PCOS MOUSE MODEL

Abstract

Polycystic ovary syndrome (PCOS) is the most common metabolic and endocrine disorder in reproductive-age women. Excessive inflammation and increased androgen production from ovarian theca cells are key features of PCOS. We and others have recently reported that human bone marrow mesenchymal stem cells (MSC) can reverse PCOS conditions both in vitro and in vivo models. Recent studies reported that MSC secreted extracellular vesicles (EV), also known as exosome, play a key role in the reparative paracrine effect of MSC in various disease. Here we evaluated the therapeutic efficacy of MSC-EV in both in vitro and in an in vivo PCOS mouse models. We hypothesize that MSC-EV treatment can reverse the PCOS related features such as insulin resistance, altered androgen production, and infertility. We used commercialized MSC-EV (EV-pure+, Vitti lab) which isolated from Human Umbilical Cord Tissue derived MSC. For in vitro experiment, we treated MSC-EV to androgen producing H293R cells, and analyzed androgen producing gene expression. For in vivo experiment, three different concentration of MSC-EV were injected into Letrozole (LTZ) induced mouse PCOS model through intravenous injection (3X107, 1.5X108, 7.5 X108 particles). Effect in both in vitro and in vivo model were assessed by monitoring cell proliferation (immunohistochemistry), steroidogenic gene expression (qRT-PCR), animal tissue assay (H&E staining), and fertility by mating and pregnancy/delivery outcomes. MSC-EV significantly reduced gene expression Cyp11a1 (0.75±0.03 fold), Cyp17a1 (0.58±0.04 fold), and Dennd1 (0.72±0.06 fold) in H293R cells. In our in vivo model, abnormal serum glucose level in PCOS mice (231.5±16.3 mg/dL) were significantly decreased by all MSC-EV tested doses treatment (186.5±3.54, 194.0±0.00, 188.0±5.66 mg/dL) at same time point after glucose injection (60 min). In breeding experiment, PCOS mice (n=4) delivered only one offspring while healthy mice (n=4) delivered 33 pups. Interestingly, average number of pups in each MSC-EV treated group(n=4) were 13, which is significantly increased compared to untreated PCOS mice. The pups did not show any visible abnormalities and exhibited normal growth rate. Our study demonstrates the efficacy of intra venous injection of MSC-EV for potential treatment of infertility in PCOS mouse model. Our work suggests that MSC-EV can potentially be a safer therapeutic option for women with PCOS as cell-free biomedicine. Further preclinical and pilot clinical trials are required to further evaluate and validate this novel treatment option for this common female reproductive disorder.