Abstract 35: 2-deoxy Adenosine Triphosphate Improves Contraction In Human End-stage Heart Failure

Abstract

We are developing a novel approach to treat heart failure that involves adeno-associated virus vectors to elevate intracellular 2 deoxy-ATP (dATP) via increased expression of the enzyme Ribonucleotide Reductase. Our studies in rodents have shown that substitution of dATP for ATP as the energy substrate increases contraction in striated muscle. Here we report for the first time the effect of dATP on human cardiac muscle contraction. We measured the contractile properties of demembranated multicellular ventricular wall preparations and isolated myofibrils from adult human heart tissue obtained from twelve patients undergoing surgery for cardiac transplantation or placement of left ventricular-assist device for end-stage heart failure. Isometric force at saturating calcium concentration was increased by about ten percent from 38.6±3.8 mN/mm2 to 42.8±4.2 mN/mm2 when dATP was substituted for ATP (P<0.001). The effect was even greater at physiologic calcium concentrations with an approximate increase of thirty percent. Isometric force increased from 21.2 ± 6.1 mN/mm2 to 26.4± 6.4 mN/mm2 (p<0.001) at pCa=5.6 and from 22.6 ± 5.9 mN/mm2 to 27.5± 6.3 mN/mm2 (p<0.001) at pCa=5.8 with dATP for ATP substitution. The result was an increase in the Ca2+ sensitivity of force as the [Ca2+] required to elicit half maximum force (pCa50) increased by 0.08 units from 5.68±0.03 to 5.79±0.03 (N=24, P< 0.001). The maximum rate of force redevelopment (ktr) in demembranated wall muscle increased (0.82±0.01 s-1 vs. 0.62±0.01 s-1, P< 0.05), as was the rate of contractile activation in isolated myofibrils (0.80±0.06 s-1 vs. 0.57±0.06 s-1, P<0.01) suggesting dATP may increase dP/dT in failing human myocardium. Importantly, there was no slowing of relaxation, as the time to 50% and 90% myofibril relaxation were unchanged. Purified myosin from failing human myocardium showed enhanced NTPase activity with dATP (0.89±0.17 s-1/head) vs. ATP (0.55±0.20 s-1/head, P<0.05). In conclusion, the data strongly suggest dATP increases cross-bridge cycling, compared with ATP in failing human myocardium and shows promise in restoring cardiac pump function. These data support a novel myofilament approach for treating heart failure that warrants further pre-clinical evaluation.