Gene transfer of calcium-binding proteins into adult cardiac myocytes

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 Ca ²⁺ binding protein motifs to restore normal Ca ²⁺ handling function to diseased myocardium. Gene transfer of parvalbumin into adult cardiac myocytes has been studied as a potential therapeutic, specifically as a strategic Ca ²⁺ 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.