Calsequestrin 2 (CASQ2) mutations increase expression of calreticulin and ryanodine receptors, causing catecholaminergic polymorphic ventricular tachycardia.
Catecholamine-induced polymorphic ventricular tachycardia (CPVT) is a familial disorder caused by cardiac ryanodine receptor type 2 (RyR2) or calsequestrin 2 (CASQ2) gene mutations. To define how CASQ2 mutations cause CPVT, we produced and studied mice carrying a human D307H missense mutation (CASQ(307/307)) or a CASQ2-null mutation (CASQ(DeltaE9/DeltaE9)). Both CASQ2 mutations caused identical consequences. Young mutant mice had structurally normal hearts but stress-induced ventricular arrhythmias; aging produced cardiac hypertrophy and reduced contractile function. Mutant myocytes had reduced CASQ2 and increased calreticulin and RyR2 (with normal phosphorylated proportions) but unchanged calstabin levels, as well as reduced total sarcoplasmic reticulum (SR) Ca(2+), prolonged Ca(2+) release, and delayed Ca(2+) reuptake. Stress further diminished Ca(2+) transients, elevated cytosolic Ca(2+), and triggered frequent, spontaneous SR Ca(2+) release. Treatment with Mg(2+), a RyR2 inhibitor, normalized myocyte Ca(2+) cycling and decreased CPVT in mutant mice, indicating RyR2 dysfunction was critical to mutant CASQ2 pathophysiology. We conclude that CPVT-causing CASQ2 missense mutations function as null alleles. In the absence of CASQ2, calreticulin, a fetal Ca(2+)-binding protein normally downregulated at birth, remains a prominent SR component. Adaptive changes to CASQ2 deficiency (increased posttranscriptional expression of calreticulin and RyR2) maintained electrical-mechanical coupling, but increased RyR2 leakiness, a paradoxical response further exacerbated by stress. The central role of RyR2 dysfunction in CASQ2 deficiency unifies the pathophysiologic mechanism underlying CPVT due to RyR2 or CASQ2 mutations and suggests a therapeutic approach for these inherited cardiac arrhythmias.
Song
Lei
L
Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Alcalai
Ronny
R
Arad
Michael
M
Wolf
Cordula M
CM
Toka
Okan
O
Conner
David A
DA
Berul
Charles I
CI
Eldar
Michael
M
Seidman
Christine E
CE
Seidman
J G
JG
D000818:Animals; D002118:Calcium / Q000378:metabolism; D037282:Calreticulin / Q000378:metabolism*; D002155:Calsequestrin / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D002395:Catecholamines / Q000096:biosynthesis*; D004594:Electrophysiology; D005091:Exons / Q000235:genetics; D017353:Gene Deletion; D005786:Gene Expression Regulation; D008274:Magnesium / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D012313:RNA / Q000235:genetics; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D017180:Tachycardia, Ventricular / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*
D037282:Calreticulin; D002155:Calsequestrin; D002395:Catecholamines; D019837:Ryanodine Receptor Calcium Release Channel; C514388:casq2 protein, mouse; D012313:RNA; D008274:Magnesium; D002118:Calcium