Comprehensive Effects of Suppression of MicroRNA-383 in Human Bone-Marrow-Derived Mesenchymal Stem Cells on Treating Spinal Cord Injury

Abstract

Background/Aims: Transplantation of bone-marrow-derived mesenchymal stem cells (MSCs) promotes neural cell regeneration after spinal cord injury (SCI). Recently, we showed that suppression of microRNA-383 (miR-383) in MSCs increased the protein levels of glial cell line derived neurotrophic factor (GDNF), resulting in improved therapeutic effects on SCI. However, the overall effects of miR-383 suppression in MSCs on SCI therapy were not determined yet. Here, we addressed this question. Methods: We used bioinformatics tools to predict all miR-383-targeting genes, confirmed the functional bindings in a dual luciferase reporter assay. The effects of alteration of candidate genes in MSCs on cell proliferation were analyzed by MTT assay and by Western blotting for PCNA. The effects on angiogenesis were assessed by HUVEC assay. The effects on SCI in vivo were analyzed by transplantation of the modified MSCs into nude rats that underwent SCI. Results: Suppression of miR-383 in MSCs not only upregulated GDNF protein, but also increased vascular endothelial growth factor A (VEGF-A) and cyclin-dependent kinase 19 (CDK19), two other miR-383 targets. MiR-383-suppression-induced increases in CDK19 resulted in a slight but significant increase in MSC proliferation, while miR-383-suppression-induced increases in VEGF-A resulted in a slight but significant increase in MSC-mediated angiogenesis. Conclusions: Upregulation of CDK19 and VEGF-A by miR-383 suppression in MSCs further improve the therapeutic potential of MSCs in treating SCI in rats.

MicroRNAs in Tumor Endothelial Cells: Regulation, Function and Therapeutic Applications

“…For instance, miR-153-3p [43], miR-383-5p [81] and miR-526b-3p [87] have been reported to target VEGF, while miR-944 targets VEGFC in ECs [100]. Notably, the direct binding of miR-383-5p to VEGF mRNA has been confirmed by Han et al in ECs [137] and demonstrated in other cell types, including bone-marrow-derived mesenchymal stem cells [138] and lung adenocarcinoma cells [139]. Moreover, miR-2355-5p (miR-2355) targets VEGFR2 in ECs [68].…”

Effects of microRNAs on angiogenesis in diabetic wounds

“…MiR-383 directly targets VEGFA and has been implicated as a regulator of angiogenesis in the highly vascular central nervous system tumor malignant glioma, with overexpression of miR-383 resulting in profound downregulation of proliferation, migration, and tube formation of cellular models of malignant glioma ( 171 ). Downregulation of miR-383 also resulted in improved angiogenesis in models of spinal cord injury in rats ( 172 ). Han et al ( 173 ) explored the effects of miR-383 in diabetic wound healing by studying its interactions with the long non-coding RNA krüppel-like factor 3 antisense RNA 1 (KLF3-AS1) delivered by BMSC-derived exosomes, and they found that these exosomes promoted increased proliferation, migration, and tube formation in HUVECs grown in high glucose media.…”

A Concise Review of MicroRNA-383: Exploring the Insights of Its Function in Tumorigenesis

“…MiR-383 suppression increased the proliferation of MSCs and MSC-mediated angiogenesis due to the increased expression of CDK-19 and VEGF-A, respectively. The up-regulated expression of CDK19 and VEGF-A further improved the therapeutic potential of MSCs in treating spinal cord injury (SCI) in rats [76].…”