Heat shock protein A12B gene therapy improves perfusion, promotes neovascularization, and decreases fibrosis in a murine model of hind limb ischemia

Mahesh Thirunavukkarasu; Muhammad Tipu Rishi; Seetur R Pradeep; Santosh Swaminathan; Diego Accorsi; John Alexander Palesty; Nilanjana Maulik

doi:10.1016/j.surg.2021.05.006

Heat shock protein A12B gene therapy improves perfusion, promotes neovascularization, and decreases fibrosis in a murine model of hind limb ischemia

Surgery. 2021 Sep;170(3):969-977. doi: 10.1016/j.surg.2021.05.006. Epub 2021 Jun 4.

Authors

Mahesh Thirunavukkarasu¹, Muhammad Tipu Rishi², Seetur R Pradeep¹, Santosh Swaminathan², Diego Accorsi², John Alexander Palesty³, Nilanjana Maulik⁴

Affiliations

¹ Molecular Cardiology and Angiogenesis Laboratory, Department of Surgery, University of Connecticut School of Medicine, Farmington, CT.
² Molecular Cardiology and Angiogenesis Laboratory, Department of Surgery, University of Connecticut School of Medicine, Farmington, CT; Stanley J. Dudrick Department of Surgery, Saint Mary's Hospital, Waterbury, CT.
³ Stanley J. Dudrick Department of Surgery, Saint Mary's Hospital, Waterbury, CT.
⁴ Molecular Cardiology and Angiogenesis Laboratory, Department of Surgery, University of Connecticut School of Medicine, Farmington, CT. Electronic address: nmaulik@uchc.edu.

PMID: 34092373
DOI: 10.1016/j.surg.2021.05.006

Abstract

Background: Heat shock protein A12B expressed in endothelial cells is important and required for angiogenesis to form functional vessels in ischemic tissue. We have previously shown the cardioprotective effects of heat shock protein A12B overexpression in a rat model of diabetic myocardial infarction. In this study, we aim to explore the role of heat shock protein A12B in a surgically-induced murine hind-limb ischemia model.

Materials and methods: Adult 8- to 12-week-old C57BL/6J mice were divided into 2 groups: treated with Adeno.LacZ (control group) and with Adeno.HSPA12B (experimental group) and, with both groups subjected to right femoral artery ligation. Immediately after surgery, mice in both groups received either Adeno.HSPA12B or Adeno.LacZ (1 × 10⁹ plaque forming units) in both the semimembranosus and gastrocnemius muscles of the right limb. The left limb served as the internal control. Both groups underwent serial laser Doppler imaging preoperatively, and again postoperatively until 28 days. Immunohistochemical analysis was performed 3 and 28 days post-surgery.

Results: Mice in the Adeno.HSPA12B gene therapy group showed improved motor function and a significantly higher blood perfusion ratio on postoperative days 21 and 28, along with better motor function. Immunohistochemical analysis showed increased expression of vascular endothelial growth factor, thioredoxin-1, heme oxygenase, and hypoxia-inducible factor 1α, along with a decreased expression of A-kinase-anchoring protein 12 and thioredoxin-interacting protein levels. The Adeno.HSPA12B-treated group also showed increased capillary and arteriolar density and an increased capillary-myocyte ratio, along with reduced fibrosis compared to the Adeno.LacZ group.

Conclusion: Our study demonstrates that targeted Adeno.HSPA12B gene delivery into ischemic muscle enhances perfusion and angiogenic protein expression. This molecule shows promise for the management of peripheral vascular disease as a potential target for clinical trials and subsequent drug therapy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Disease Models, Animal
Fibrosis / prevention & control
Genetic Therapy / methods*
HSP70 Heat-Shock Proteins / genetics*
HSP70 Heat-Shock Proteins / physiology
Hindlimb / blood supply
Ischemia / therapy*
Male
Mice
Mice, Inbred C57BL
Motor Activity
Neovascularization, Physiologic*
Regional Blood Flow*

Substances

HSP70 Heat-Shock Proteins
HSPA12B protein, mouse