Effects of genetic transfection on calcium cycling pathways mediated by double-stranded adeno-associated virus in postinfarction remodeling

J Thorac Cardiovasc Surg. 2020 May;159(5):1809-1819.e3. doi: 10.1016/j.jtcvs.2019.08.089. Epub 2019 Sep 30.

Abstract

Objective: Restoring calcium sensor protein (S100A1) activity in failing hearts poses a promising therapeutic strategy. We hypothesize that cardiac overexpression of the S100A1 gene mediated by a double-stranded adeno-associated virus (scAAV) results in better functional and molecular improvements compared with the single-stranded virus (ssAAV).

Methods: Heart failure was induced by coronary artery ligation. Then, intramyocardial injections of saline, AAV9 empty capsid, scAAV9.S100A1, and ssAAV9.S100A1 were performed. Ten weeks postinfarction, all rats received cardiac evaluation; serum and tissue were collected for genetic analysis, cytokine profiling, and assessments of mitochondrial function and structure.

Results: Overexpression of AAV9.S100A1 improved systolic and diastolic function. Compared with control, ejection fraction was greater in treated groups (54.8% vs 32.3%, P < .05). Similarly, end-diastolic volume index was significantly less in the treated group than in control (1.14 vs 1.59 mL/cm2), whereas fractional shortening was greater in treated groups than control (26% vs 38%, P < .05). Interestingly, cardiac mechanics were significantly better when treated with double-stranded virus compared with single-stranded. Quantitative polymerase chain reaction demonstrated robust transfection of myocardium with the S100A1 gene, with more infection in the self-complimentary group compared with the single-stranded group (5.68 ± 0.44 vs 4.09 ± 0.25 log10 genome copies per 100 ng of DNA; P < .0001). Concentrations of the inflammatory cytokines were elevated in the ssAAV9/S100A1 group compared with the scAAV9/S100A1. Assessment of mitochondrial respiration and morphology demonstrated that injection of self-complementary vector saved both mitochondrial structure and function.

Conclusions: Gene therapy of S100A1 can prevent pathologic postmyocardial infarction remodeling and decrease inflammatory response in ischemic heart failure.

Keywords: adeno-associated virus; cytokines; gene therapy; inflammation; mitochondria; post-myocardial remodeling.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Animals
  • Calcium Signaling*
  • Cytokines / metabolism
  • Dependovirus / genetics*
  • Dependovirus / metabolism
  • Disease Models, Animal
  • Fibrosis
  • Genetic Therapy*
  • Genetic Vectors*
  • Heart Failure / genetics
  • Heart Failure / metabolism
  • Heart Failure / physiopathology
  • Heart Failure / therapy*
  • Heart Ventricles / metabolism*
  • Heart Ventricles / pathology
  • Heart Ventricles / physiopathology
  • Inflammation Mediators / metabolism
  • Lipid Peroxidation
  • Male
  • Mitochondria, Heart / metabolism
  • Mitochondria, Heart / pathology
  • Myocardial Infarction / genetics
  • Myocardial Infarction / metabolism
  • Myocardial Infarction / physiopathology
  • Myocardial Infarction / therapy*
  • Rats, Sprague-Dawley
  • Reactive Oxygen Species / metabolism
  • S100 Proteins / biosynthesis
  • S100 Proteins / genetics*
  • Stroke Volume
  • Transfection*
  • Ventricular Function, Left*
  • Ventricular Remodeling*

Substances

  • Cytokines
  • Inflammation Mediators
  • Reactive Oxygen Species
  • S100 Proteins
  • S100A1 protein