Thin Filament Disinhibition by Restrictive Cardiomyopathy Mutant R193H Troponin I Induces Ca -Independent Mechanical Tone and Acute Myocyte Remodeling

Inherited restrictive cardiomyopathy (RCM) is a debilitating disease characterized by a stiff heart with impaired ventricular relaxation. Mutations in cardiac troponin I (cTnI) were identified as causal for RCM. Acute genetic engineering of adult cardiac myocytes was used to identify primary structure/function effects of mutant cTnI. Studies focused on R193H cTnI owing to the poor prognosis of this allele. Compared with wild-type cTnI, R193H mutant cTnI more effectively incorporated into the sarcomere, where it exerted dose-dependent effects on basal and dynamic contractile function. Under loaded conditions, permeabilized myocyte Ca sensitivity of tension was increased, whereas the passive tension–extension relationship was not altered by R193H cTnI. Normal rod-shaped myocyte morphology acutely transitioned to a “short-squat” phenotype in concert with progressive stoichiometric incorporation of R193H in the absence of altered diastolic Ca . The specific myosin inhibitor blebbistatin fully blocked this transition. Heightened Ca buffering by the R193H myofilaments, and not alterations in Ca handling by the sarcoplasmic reticulum, slowed the decay rate of the Ca transient. Incomplete mechanical relaxation conferred by R193H was exacerbated at increasing pacing frequencies independent of elevated diastolic Ca . R193H cTnI–dependent mechanical tone caused acute remodeling to a quasicontracted state not elicited by other Ca -sensitizing proteins and is a direct correlate of the stiff heart characteristic of RCM in vivo. These results point toward targets downstream of Ca handling, notably thin filament regulation and actin–myosin interaction, in designing therapeutic strategies to redress the primary cell morphological and mechanical underpinnings of RCM. (Circ Res. 2007;100:1494-1502.)