PMID- 37645966
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20240210
DP  - 2023 Aug 14
TI  - Inhibiting Anti-angiogenic VEGF165b Activates a Novel miR-17-20a-Calcipressin-3 
      Pathway that Revascularizes Ischemic Muscle in Peripheral Artery Disease.
LID - rs.3.rs-3213504 [pii]
LID - 10.21203/rs.3.rs-3213504/v1 [doi]
AB  - BACKGROUND: VEGF(165)a increases the expression of microRNA-17-92 cluster, 
      promoting developmental, retinal, and tumor angiogenesis. We have previously 
      shown that VEGF(165)b, an alternatively spliced VEGF-A isoform, inhibits the 
      VEGFR-STAT3 pathway in ischemic endothelial cells (ECs) to decrease their 
      angiogenic capacity. In ischemic macrophages (Mos), VEGF(165)b inhibits VEGFR1 to 
      induce S100A8/A9 expression, which drives M1-like polarization. Our current study 
      aims to determine whether VEGF(165)b inhibition promotes perfusion recovery by 
      regulating the miR-17-92 cluster in preclinical PAD. METHODS: Hind limb ischemia 
      (HLI) induced by femoral artery ligation and resection was used as a preclinical 
      PAD model. Hypoxia serum starvation (HSS) was used as an in vitro PAD model. 
      VEGF(165)b was inhibited/neutralized by an isoform-specific VEGF(165)b antibody. 
      RESULTS: Systematic analysis of miR-17-92 cluster members 
      (miR-17-18a-19a-19b-20a-92) in experimental-PAD models showed that 
      VEGF(165)b-inhibition induces miRNA-17-20a (within miR-17-92 cluster) in HSS-ECs 
      and HSS-bone marrow derived macrophages (BMDMs) vs. respective normal and/or 
      isotype matched IgG controls to enhance perfusion-recovery. Consistent with the 
      bioinformatics analysis that revealed RCAN3 as a common target of miR-17 and 
      miR-20a, Argonaute-2 pull-down assays showed decreased miR-17-20a expression and 
      higher RCAN3 expression in the RISC complex of HSS-ECs and HSS-BMDMs vs. the 
      respective controls. Inhibiting miR-17-20a induced RCAN3 levels to decrease 
      ischemic angiogenesis and promoted M1-like polarization to impair perfusion 
      recovery. Finally, using STAT3 inhibitors, S100A8/A9 silencers and 
      VEGFR1-deficient ECs and Mos, we show that VEGF(165)b inhibition activates the 
      miR-17-20a-RCAN3 pathway independent of VEGFR1-STAT3 or VEGFR1-S100A8/A9 in 
      ischemic ECs and ischemic Mos, respectively. CONCLUSION: Our data revealed a 
      hereunto unrecognized therapeutic 'miR-17-20a-RCAN3' pathway in the ischemic 
      vasculature that is VEGFR1-STAT3/S100A8/A9 independent and is activated only upon 
      VEGF(165)b inhibition in PAD.
FAU - Batan, S
AU  - Batan S
AD  - Vascular Biology Center, Department of Medicine, Augusta University, 
      Augusta-GA-30912.
FAU - Kuppuswamy, S
AU  - Kuppuswamy S
AD  - Vascular Biology Center, Department of Medicine, Augusta University, 
      Augusta-GA-30912.
FAU - Wood, M
AU  - Wood M
AD  - Medical College of Georgia, Augusta University, Augusta-GA-30912.
FAU - Reddy, M
AU  - Reddy M
AD  - Medical College of Georgia, Augusta University, Augusta-GA-30912.
FAU - Annex, B H
AU  - Annex BH
AD  - Vascular Biology Center, Department of Medicine, Augusta University, 
      Augusta-GA-30912.
FAU - Ganta, V C
AU  - Ganta VC
AD  - Vascular Biology Center, Department of Medicine, Augusta University, 
      Augusta-GA-30912.
LA  - eng
GR  - R01 HL141325/HL/NHLBI NIH HHS/United States
GR  - R01 HL150003/HL/NHLBI NIH HHS/United States
GR  - R01 HL146673/HL/NHLBI NIH HHS/United States
GR  - R01 HL101200/HL/NHLBI NIH HHS/United States
GR  - R01 HL148590/HL/NHLBI NIH HHS/United States
GR  - R01 GM129074/GM/NIGMS NIH HHS/United States
PT  - Preprint
DEP - 20230814
PL  - United States
TA  - Res Sq
JT  - Research square
JID - 101768035
UIN - Commun Med (Lond). 2024 Jan 5;4(1):3. PMID: 38182796
PMC - PMC10462251
OTO - NOTNLM
OT  - Angiogenesis
OT  - Ischemia
OT  - Macrophage polarization
OT  - OncomiR
OT  - VEGF-A isoforms
COIS- Competing interests: The authors declare no conflict of interest in the study.
EDAT- 2023/08/30 06:48
MHDA- 2023/08/30 06:49
PMCR- 2023/08/28
CRDT- 2023/08/30 04:03
PHST- 2023/08/30 06:48 [pubmed]
PHST- 2023/08/30 06:49 [medline]
PHST- 2023/08/30 04:03 [entrez]
PHST- 2023/08/28 00:00 [pmc-release]
AID - rs.3.rs-3213504 [pii]
AID - 10.21203/rs.3.rs-3213504/v1 [doi]
PST - epublish
SO  - Res Sq [Preprint]. 2023 Aug 14:rs.3.rs-3213504. doi: 10.21203/rs.3.rs-3213504/v1.