Neprilysin inhibition reduces microtubule detyrosination in cardiomyocytes through a cGMP-PRKG1-VASH1 axis

Abstract Microtubule detyrosination and re-tyrosination on the C-terminus of α-tubulin are mediated by the vasohibin (VASH)-small vasohibin-binding protein (SVBP) complex and tubulin tyrosine ligase (TTL), respectively. Elevated levels of detyrosinated α-tubulin (dTyr-tub) are observed in heart failure, and reducing this modification improves cardiac function, suggesting that heart failure therapies may modulate microtubule detyrosination. We investigated whether sacubitrilat and valsartan, the active components of the angiotensin receptor–neprilysin inhibitor LCZ696, influence dTyr-tub levels in endothelin-1 (ET1)-induced hypertrophy in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). While both sacubitrilat and valsartan prevented hypertrophy, only sacubitrilat prevented ET1-induced dTyr-tub accumulation. Sacubitrilat slightly increased cyclic guanosine 3′,5′-monophosphate (cGMP) levels and lowered dTyr-tub, whereas inhibition or knockdown of the cGMP-dependent protein kinase 1 (PRKG1) increased dTyr-tub. PRKG1 phosphorylated native VASH1 . Incubation of microtubules with the VASH1-SVBP complex containing wild-type VASH1, but not a VASH1 phosphomimic (VASH1-7E) increased detyrosination. Consistently, overexpression of VASH1-7E gave rise to lower dTyr-tub level than overexpression of a non-phosphorylatable form of VASH1 (VASH1-7A) in hiPSC-CMs deficient in VASH1. These findings identify a cGMP–PRKG1–VASH1 signaling axis that reduces microtubule detyrosination in cardiomyocytes.