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Loss of m6A demethylase ALKBH5 promotes post-ischemic angiogenesis via post-transcriptional stabilization of WNT5A
- Source :
- Clinical and Translational Medicine, Vol 11, Iss 5, Pp n/a-n/a (2021), Clinical and Translational Medicine
- Publication Year :
- 2021
-
Abstract
- Background Post‐ischemic angiogenesis is critical for blood flow recovery and ischemic tissue repair. N6‐methyladenosine (m6A) plays essential roles in numerous biological processes. However, the impact and connected mechanism of m6A on post‐ischemic angiogenesis are not fully understood. Methods AlkB homolog 5 (ALKBH5) was screened out among several methyltransferases and demethylases involved in dynamic m6A regulation. Cardiac microvascular endothelial cells (CMECs) angiogenesis and WNT family member 5A (WNT5A) stability were analyzed upon ALKBH5 overexpression with adenovirus or knockdown with small interfering RNAs in vitro. The blood flow recovery, capillary, and small artery densities were evaluated in adeno‐associated virus (AAV)‐ALKBH5 overexpression or ALKBH5 knockout (KO) mice in a hind‐limb ischemia model. The same experiments were conducted to explore the translational value of transient silencing of ALKBH5 with adenovirus. Results ALKBH5 was significantly upregulated in hypoxic CMECs and led to a global decrease of m6A level. ALKBH5 overexpression further reduced m6A level in normoxic and hypoxic CMECs, impaired proliferation, migration, and tube formation only in hypoxic CMECs. Conversely, ALKBH5 knockdown preserved m6A levels and promoted angiogenic phenotypes in hypoxic but not in normoxic CMECs. Mechanistically, ALKBH5 regulated WNT5A expression through post‐transcriptional mRNA modulation in an m6A‐dependent manner, which decreased its stability and subsequently impeded angiogenesis in hypoxic CMECs. Furthermore, ALKBH5 overexpression hindered blood flow recovery and reduced CD31 and alpha‐smooth muscle actin expression in hind‐limb ischemia mice. As expected, ALKBH5‐KO mice exhibited improved blood flow recovery, increased capillary, and small artery densities after hind‐limb ischemia, and similar beneficial effects were observed in mice with transient adenoviral ALKBH5 gene silencing. Conclusion We demonstrate that ALKBH5 is a negative regulator of post‐ischemic angiogenesis via post‐transcriptional modulation and destabilization of WNT5A mRNA in an m6A‐dependent manner. Targeting ALKBH5 may be a potential therapeutic option for ischemic diseases, including peripheral artery disease.
- Subjects :
- 0301 basic medicine
CD31
Medicine (General)
Angiogenesis
Ischemia
Medicine (miscellaneous)
Wnt-5a Protein
03 medical and health sciences
Mice
0302 clinical medicine
R5-920
Downregulation and upregulation
peripheral arterial disease
medicine
Gene silencing
Animals
Humans
RNA, Messenger
RNA Processing, Post-Transcriptional
Research Articles
Tube formation
Gene knockdown
Neovascularization, Pathologic
Chemistry
Wnt signaling pathway
AlkB homolog 5
AlkB Homolog 5, RNA Demethylase
medicine.disease
Cell biology
Hindlimb
Up-Regulation
Disease Models, Animal
030104 developmental biology
030220 oncology & carcinogenesis
N6‐methyladenosine
Gene Knockdown Techniques
Blood Circulation
Molecular Medicine
Research Article
post‐ischemic angiogenesis
Subjects
Details
- ISSN :
- 20011326
- Volume :
- 11
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Clinical and translational medicine
- Accession number :
- edsair.doi.dedup.....caac47156c0df8b50166a6492df6b2be