Human Stem Cells Overexpressing miR-21 Promote Angiogenesis in Critical Limb Ischemia by Targeting CHIP to Enhance HIF-1α Activity

Abstract

Critical limb ischemia (CLI) is a severe blockage in the arteries of the lower extremities. However, the effective and optimal treatment for CLI remains to be elucidated. Previous therapeutic research is mainly focused on proangiogenic growth factors administrations. Recently, miR-21 has been revealed to play a crucial role in angiogenesis. Thus, we hypothesize that miR-21 over-expression in human umbilical cord blood-derived mesenchymal stem cells (UCBMSCs) can effectively treat CLI. Herein, UCBMSCs were transduced with lentivirus-miR-21-Luciferase (Lenti-miR-21) or lentivirus-LacZ-Luciferase (Lenti-LacZ). The results indicated that miR-21 induced UCBMSCs proliferation, migration, and angiogenesis in vitro. Subsequently, general observation and laser Doppler perfusion imaging were introduced to detect perfusion in muscles of CLI-nude mice on 1, 4, 7, 14, and 28 day postoperation. There was a significant improvement in blood vessels of the ischemic limb in Lenti-miR-21 group at 7 day compared with the saline or Lenti-LacZ groups. At 28 day, histological analysis confirmed that UCBMSCs over-expressing miR-21 increased neovascularization in CLI. Furthermore, carboxyl terminus of Hsc70-interacting protein (CHIP) was found to be the target gene for miR-21-mediated activation of hypoxia-inducible factor 1a (HIF-1a) in UCBMSCs. In summary, our study demonstrated that over-expressing miR-21 in UCBMSCs could improve neovascularization in CLI through enhancing HIF-1a activity by targeting CHIP, which may hold great therapeutic promise in treating CLI. STEM CELLS 2016;34:924-934 SIGNIFICANCE STATEMENTOur study demonstrated that over-expressing miR-21 in umbilical cord blood-derived mesenchymal stem cells could improve neovascularization in critical limb ischemia through enhancing hypoxia-inducible factor 1a activity by targeting carboxyl terminus of Hsc70-interacting protein, which may hold great therapeutic promise in treating critical limb ischemia diseases.

Collagen biomaterial stimulates the production of extracellular vesicles containing microRNA‐21 and enhances the proangiogenic function of CD34 + cells

“…Relevant to the present study, miR-21 has been shown to play a vital role in vessel sprouting by regulating both HIF-1a and VEGF (55). It has also been shown that overexpression of miR-21 in mesenchymal stem cells resulted in an increase in proliferation, migration, and angiogenesis in vitro and increased neovascularization in a critical limb ischemia model (67). Herein, we demonstrate that in response to biomaterial culture, miR-21 levels are elevated in both CD34 + cells and the EVs they secrete.…”

Differential and Interactive Effects of Substrate Topography and Chemistry on Human Mesenchymal Stem Cell Gene Expression

“…The local shortage of dissolved oxygen would upregulate expression levels of certain hypoxic proteins, e.g., MMP9, VEGF, MIF, PGK1, HIF1A, LDHA, TWIST, HGF, FGF, etc. [52,53,54,55,56,57]. Yet, none of these proteins were found to be upregulated in the Fs samples.…”

miR‐21 modification enhances the performance of adipose tissue‐derived mesenchymal stem cells for counteracting urethral stricture formation

“…Increasing studies also indicated that MSCs derived angiogenic proteins could regulate the neoangiogenesis injury tissue for promoted wound healing process. Recently study also proved that miR‐21 overexpressed MSCs can effectively induce angiogenesis potential via promoting HIF‐1a activity . In the present study, in order to understand the mechanism of miR‐21 modified ADSCs in counteract urethral stricture formation, we examined the expression levels of some angiogenic genes, such as VEGF, HIF‐1a and SDF‐1a.…”

“…Recently study also proved that miR-21 overexpressed MSCs can effectively induce angiogenesis potential via promoting HIF-1a activity. 24 In the present study, in order to understand the mechanism of miR-21 modified ADSCs in counteract urethral stricture formation, we examined the expres- Together, these results suggested that ADSCs transplantation could effectively improve the healing and reconstitution of the urethra tissue to counteract US formation, and the miR-21 modification via lentiviral transfection can increase the therapeutic efficacy.…”