Gene Transfer of Calcium-Binding Proteins into Adult Cardiac Myocytes

Methods Mol Biol. 2019:1929:187-205. doi: 10.1007/978-1-4939-9030-6_12.

Abstract

Heart failure is the leading cause of combined morbidity and mortality in the USA with 50% of cases being diastolic heart failure. Diastolic heart failure results from poor myocardial relaxation and inadequate filling of the left ventricular chamber caused in part by calcium-handling dysregulation. In this chapter we describe methods to investigate new approaches of novel human Ca2+ binding protein motifs to restore normal Ca2+ handling function to diseased myocardium. Gene transfer of parvalbumin into adult cardiac myocytes has been studied as a potential therapeutic, specifically as a strategic Ca2+ buffer to correct cardiac mechanical dysfunction in disease. This chapter provides protocols for studying wild-type parvalbumin isoforms and parvalbumins with strategically designed EF-hand motifs in adult cardiac myocytes via acute adenoviral gene transfer. These protocols have been used extensively to optimize parvalbumin function as a potential therapeutic for failing heart muscle.

Keywords: Adult cardiac myocyte; Calcium; Calcium imaging; Contractility; Gene transfer; Parvalbumin.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenoviridae / genetics*
  • Adult
  • Animals
  • Calcium / metabolism
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism
  • Female
  • Gene Transfer Techniques*
  • Genetic Vectors / pharmacology
  • Heart Failure / genetics
  • Heart Failure / metabolism
  • Heart Failure / physiopathology
  • Heart Ventricles / metabolism
  • Heart Ventricles / physiopathology
  • Humans
  • Models, Biological
  • Mutation
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Parvalbumins / genetics
  • Parvalbumins / metabolism*
  • Rats, Sprague-Dawley

Substances

  • Calcium-Binding Proteins
  • Parvalbumins
  • Calcium