Extracellular Vesicles From Human Urine-Derived Stem Cells Ameliorate Erectile Dysfunction in a Diabetic Rat Model by Delivering Proangiogenic MicroRNA

Abstract

Introduction: Stem cell therapies represent a promising new frontier for the treatment of refractory diabetic erectile dysfunction (DED). The use of stem cell-derived extracellular vesicles (EVs) is a novel strategy for cellfree stem cell therapy. We have reported that urine-derived stem cells (USCs) can improve DED; however, the therapeutic effects of EVs secreted by USCs (USC-EVs) remain unknown. Aim: To determine the therapeutic effects of USC-EVs on DED in a rat model. Methods: USC-EVs were isolated from conditioned medium by ultracentrifugation. DED was induced in male SpragueeDawley rats via an intraperitoneal injection of streptozotocin. Sixteen DED rats were divided into phosphate-buffered saline (PBS) and USC-EV groups. Eight normal rats served as the normal control group. PBS or USC-EVs were transplanted into the corpora cavernosa in the corresponding groups. Main Outcome Measure: Intracavernosal pressure (ICP), mean arterial pressure (MAP), expression of endothelial markers (CD31), endothelial nitric oxide synthase (eNOS), phospho-eNOS, and neural nitric oxide synthase (nNOS) were assessed in each group. Masson's trichrome staining was used to determine the collagen deposition and ratio of smooth muscle cells to collagen. The microRNA (miRNA) cargo of USC-EVs was characterized by high-throughput RNA sequencing. Results: Recovery of erectile function was observed in the USC-EV group, as represented by improved ICP and ICP/MAP ratio. CD31, eNOS, phospho-eNOS, and nNOS expression in the penis was significantly improved in the USC-EV group. In addition, the ratio of smooth muscle to collagen was significantly increased in the USC-EV group. RNA sequencing revealed that USC-EVs were enriched for distinct classes of miRNA (miR-21-5p, let-7 family, miR-10 family, miR-30 family, and miR-148a-3p) that promote angiogenesis. Conclusion: USC-EV transplantation can ameliorate DED in rats. Its mechanism may involve the delivery of proangiogenic miRNA. Ouyang B, Xie Y, Zhang C, et al. Extracellular Vesicles From Human Urine-Derived Stem Cells Ameliorate Erectile Dysfunction in a Diabetic Rat Model by Delivering Proangiogenic MicroRNA. Sex Med 2019;7:241e250.

Urine-Derived Stem Cells: Applications in Regenerative and Predictive Medicine

“…Regarding challenges and open questions, UDSC biological characterizations need to be deeper explored, with a special focus on donor age effects on UDSC properties, as previously mentioned. Moreover, in UDSC, the expression levels of some characteristic surface markers, analyzed by flow cytometry, show discrepancy among different studies [20,21,29,32,38,46,68,69,73,84,95,[98][99][100]. These discrepancies can be attributed to different culture conditions, such as the composition of the culture medium, or can be a native characteristic of the isolated cells that can be heterogeneous within the same individual or between different individuals.…”

“…Chen et al demonstrated that systemic injection of extracellular vesicles from human-harvested UDSC prevented osteoporosis in ovariectomized mice by transferring collagen triple-helix repeat containing 1 (CTHRC1) and osteoprotegerin (OPG) [71]. The same strategy was done to improve erectile dysfunction in a diabetic rat model-this time, by means of transferring pro-angiogenic miRNA [72]-and to decrease kidney injury in a diabetic rat model by transferring pro-vascular regeneration and cell protection factors, including growth factor, transforming growth factor-β1, angiogenin and bone morphogenetic protein-7 [73]. In the same disease context, Jin et al demonstrated that exosomes from UDSC attenuated podocyte apoptosis in a mouse model of diabetes, besides decreasing apoptosis and increasing autophagy fluxes when added to MPC5 cells exposed to hyperglycemia.…”

Extracellular vesicles in type 2 diabetes mellitus: key roles in pathogenesis, complications, and therapy

“…This decreased apoptosis could effectively ameliorate the erectile function in rat model of T2DM [92]. Additionally, Ouyang et al [93] found that the erectile dysfunction of diabetic rats was effectively ameliorated after the EVs of urine‐derived stem cells were transplanted into the corpora cavernosa of rats. RNA sequencing revealed that different miRNAs (miR‐21‐5p, let‐7, and miR‐10) that promote angiogenesis of corpora cavernosa were enriched in these EVs.…”

Urine as a Main Effector in Urological Tissue Engineering—A Double-Edged Sword

“…Exosome production by human UDSCs can suppress kidney complications in diabetic rats. Moreover, storing and managing EVs is much easier in comparison to stem cells, in order to decrease the chance of formation of a tumor as lower clearance of EVs enhances tumor formation [59]. Prattichizzo et al proposed that EVs can treat endothelial metabolic memory through the delivery of miRNA [60].…”

“…In a study conducted by Ouyang B et al [59], it was presented that reduce deposition of UDSC-EVs enhanced the improved neurogenic-mediated erectile reaction in diabetic erectile dysfunction and endothelial cell marker expression. The erectile response enhancement was indicated by improved ICP/MAP and ICP ratio, which was attributed to enhanced smooth muscle and endothelium function.…”