Enhancing calmodulin binding to cardiac ryanodine receptor completely inhibits pressure-overload induced hypertrophic signaling

Abstract

Cardiac hypertrophy is a well-known major risk factor for poor prognosis in patients with cardiovascular diseases. Dysregulation of intracellular Ca2+ is involved in the pathogenesis of cardiac hypertrophy. However, the precise mechanism underlying cardiac hypertrophy remains elusive. Here, we investigate whether pressure-overload induced hypertrophy can be induced by destabilization of cardiac ryanodine receptor (RyR2) through calmodulin (CaM) dissociation and subsequent Ca2+ leakage, and whether it can be genetically rescued by enhancing the binding affinity of CaM to RyR2. In the very initial phase of pressure-overload induced cardiac hypertrophy, when cardiac contractile function is preserved, reactive oxygen species (ROS)-mediated RyR2 destabilization already occurs in association with relaxation dysfunction. Further, stabilizing RyR2 by enhancing the binding affinity of CaM to RyR2 completely inhibits hypertrophic signaling and improves survival. Our study uncovers a critical missing link between RyR2 destabilization and cardiac hypertrophy.

GRP78 Antibodies Are Associated With Blood-Brain Barrier Breakdown in Anti–Myelin Oligodendrocyte Glycoprotein Antibody–Associated Disorder

“…In brief, total RNA was extracted from TY10 cells using the RNeasy Mini Kit (Qiagen), and messenger RNA was purified as described previously. 20 Complementary DNA libraries were produced using a NEBNext Ultra II RNA Library Prep kit (New England Biolabs) and NEBNextplex Oligos for Illumina, as described previously. 20 In this approach, messenger RNA was fragmented in NEBNext First Strand Synthesis Reaction Buffer at 94°C for 15 minutes in the presence of NEBNext Random Primers and was reverse transcribed with NEBNext Strand Synthesis Enzyme Mix.…”

“… 20 Complementary DNA libraries were produced using a NEBNext Ultra II RNA Library Prep kit (New England Biolabs) and NEBNextplex Oligos for Illumina, as described previously. 20 In this approach, messenger RNA was fragmented in NEBNext First Strand Synthesis Reaction Buffer at 94°C for 15 minutes in the presence of NEBNext Random Primers and was reverse transcribed with NEBNext Strand Synthesis Enzyme Mix. The library fragments were then concentrated, and index sequences were inserted during polymerase chain reaction amplification.…”

“…The data were then trimmed and mapped to the mouse reference genome GRCm38 release-92 using the CLC Genomics Workbench software program (ver.8.01; Qiagen) as described previously. 20 The mapped read counts were normalized to transcripts per million, which were converted to log2 after the addition of 1. For the volcano plots, p values were calculated with the unpaired Student t test, and the fold change (FC) was determined by subtracting the average values in the HCs from those in patients.…”

Highly concentrated trehalose induces transient senescence-associated secretory phenotype in fibroblasts via CDKN1A/p21

“…Total RNA was extracted from the fibroblasts or LSE using the RNeasy Mini Kit (Qiagen, Hilden, Germany), and mRNA was purified with oligo dT beads (NEBNext Poly (A) mRNA magnet Isolation Module, New England Biolabs, NEB, Ipswich, MA). The procedure of the complementary DNA (cDNA) libraries was carried out with NEBNext Ultra II RNA library Prep kit (NEB) and NEBNextplex Oligos for Illumina following a previously described method (Kohno et al, 2020).…”

Herpud1 suppress angiotensin II induced hypertrophy in cardiomyocytes

“…In our previous study, we reported that RyR2 V3599K KI mice, in which the calmodulin (CaM) binding domain of the myocardial ryanodine receptor (RyR2) was mutated to enhance the affinity of CaM to RyR2 [ 5 ], do not develop cardiac hypertrophy in the transverse aortic coarctation (TAC) model [ 6 ]. In this model, LV systolic and diastolic function is maintained without LV hypertrophy, even when the LV peak pressure is increased in WT mice.…”