Transcriptomic Analysis of Human Mesenchymal Stem Cell Therapy in Incontinent Rat Injured Urethra

Abstract

Periurethral human mesenchymal stem cell (hMSC) injections are associated with functional improvement in animal models of postpartum stress urinary incontinence (SUI). However, limited data exist on the role of hMSCs in modulating gene expression in tissue repair after urethral injury. To this end, we quantified temporal gene expression modulation in hMSCs, and in injured rat urethral tissue, using RNA-seq in an animal model of SUI, over a 3-day period following urethral injury, and local hMSC injection. We injected PKH fluorescent-labeled hMSC into the periurethral space of rats following a 4 h vaginal distention (VD) (three rats per time point). Control rats underwent VD injury only, and all animals were euthanized at 12, 24, 36, 72 h postinjury. Rat urethral and vaginal tissues were frozen and sectioned. Fluorescent labeled hMSCs were distinguished from adjacent, unlabeled rat urethral tissue. RNA was prepared from hMSCs and urethral tissue obtained by laser dissection of frozen tissue sections and sequenced on an Illumina HiSeq 2500. Differentially expressed genes (DEGs) over 72 h were evaluated using a two-group t-test (p < 0.05). Our transcriptional analyses identified candidate genes involved in tissue injury that were broadly sorted by injury and exposure to hMSC throughout the first 72 h of acute phase of injury. DEGs in treated urethra, compared with untreated urethra, were functionally associated with tissue repair, angiogenesis, neurogenesis, and oxidative stress suppression. DEGs included a variety of cytokines, extracellular matrix stabilization and regeneration genes, cytokine signaling modification, cell cycle regulation, muscle differentiation, and stabilization. Moreover, our results revealed DEG changes in hMSCs (PKH-labeled) harvested from injured urethra. The expressions are related to DNA damage repair, transcription activation, stem cell regulation, cell survival, apoptosis, self-renewal, cell proliferation, migration, and injury response.

Mesenchymal stem cell-based therapy for female stress urinary incontinence

“…Reasons for the different outcomes might be attributed to the cell source, the culture environment in vitro, and the type of receptors. To test the hypothesis that long-term paracrine effects of hMSCs reduce inflammatory responses and modulate a healing microenvironment, Sadeghi et al (2020) performed the first all-round transcriptomic analysis of hMSC therapy for postpartum SUI. The authors observed various differentially expressed genes in the injured urethra treated with hMSCs related to several signaling pathways involved in tissue regeneration and repair, such as wound healing, vasoprotection, angiogenesis, neurogenesis, and neuroprotection as well as oxidative stress inhibition.…”

“…Although the feasibility of MSC therapy for SUI has been demonstrated (Kim et al, 2012;Silwal Gautam et al, 2014;Sadeghi et al, 2016;Garcia-Arranz et al, 2020;Sadeghi et al, 2020), MSCs alone may be insufficient to regenerate whole urethral components. Table 2 lists the features of MSC-based combination therapies for SUI.…”

Identification of key pathways and hub genes in the myogenic differentiation of pluripotent stem cell: a bioinformatics and experimental study

“…[5][6][7]. Stem cells are promising cells that have the potency of multidirectional differentiation and proliferation and are widely expected to be used in the field of tissue repair and regeneration [8,9]. In the muscle regeneration field, stem cells also showed vigorous potency [10].…”

Identification of key pathways and hub genes in the myogenic process of pluripotent stem cell: A bioinformatics study

“…Stem cells are promising cells that have the potency of multi-directional differentiation and proliferation and are widely expected to be used in the eld of tissue repair and regeneration. [8,9] In the muscle regeneration eld, stem cells also showed vigorous potency. [10] Various researches have devoted to verifying the mechanism of myoblast differentiation.…”