Hypertrophic cardiomyopathy (HCM) is the most common inherited heart disease and is characterized by primary left ventricular hypertrophy usually caused by mutations in sarcomere genes. The mechanism underlying cardiac remodeling in HCM remains incompletely understood. An investigation of HCM through integrative analysis at multi-omics levels will be helpful for treating HCM. DNA methylation and chromatin accessibility, as well as gene expression, were assessed by nucleosome occupancy and methylome sequencing (NOMe-seq) and RNA-seq, respectively, using the cardiac tissues of HCM patients. Compared with those of the controls, the transcriptome, DNA methylome, and chromatin accessibility of the HCM myocardium showed multifaceted differences. At the transcriptome level, HCM hearts returned to the fetal gene program through decreased sarcomeric and metabolic gene expression and increased extracellular matrix gene expression. In the DNA methylome, hypermethylated and hypomethylated differentially methylated regions were identified in HCM. At the chromatin accessibility level, HCM hearts showed changes in different genome elements. Several transcription factors, including SP1 and EGR1, exhibited a fetal-like pattern of binding motifs in nucleosome-depleted regions in HCM. In particular, the inhibition of SP1 or EGR1 in an HCM mouse model harboring sarcomere mutations markedly alleviated the HCM phenotype of the mutant mice and reversed fetal gene reprogramming. Overall, this study not only provides a high-precision multi-omics map of HCM heart tissue but also sheds light on the therapeutic strategy by intervening in the fetal gene reprogramming in HCM.
Cardiomyopathy, Hypertrophic/metabolism* ; Humans ; Animals ; DNA Methylation ; Mice ; Transcriptome ; Chromatin/genetics* ; Early Growth Response Protein 1/metabolism* ; Male ; Epigenome ; Nucleosomes/genetics* ; Female ; Middle Aged ; Disease Models, Animal ; Adult

Noonan syndrome (NS) is an autosomal dominant disorder that involves multiple organ systems, with short stature as the most common presentation (>70%). Possible mechanisms of short stature in NS include growth hormone (GH) deficiency, neurosecretory dysfunction, and GH resistance. Accordingly, GH therapy has been carried out for NS patients over the last three decades, and multiple studies have reported acceleration of growth velocity (GV) and increase of height standard deviation score (SDS) in both prepubertal and pubertal NS patients upon GH therapy. One year of GH therapy resulted in almost doubling of GV compared with baseline; afterwards, the increase in GV gradually decreased in the following years, showing that the effect of GH therapy wanes over time. After four years of GH therapy, ~70% of NS patients reached normal height considering their age and sex. Early initiation, long duration of GH therapy, and higher height SDS at the onset of puberty were associated with improved final height, whereas gender, dosage of GH, and the clinical severity did not show significant association with final height. Studies have reported no significant adverse events of GH therapy regarding progression of hypertrophic cardiomyopathy, alteration of metabolism, and tumor development. Therefore, GH therapy is effective for improving height and GV of NS patients; nevertheless, concerns on possible malignancy remains, which necessitates continuous monitoring of NS patients receiving GH therapy.
Acceleration ; Adolescent ; Cardiomyopathy, Hypertrophic ; Growth Hormone* ; Humans ; Metabolism ; Noonan Syndrome* ; Puberty

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; pharmacology ; Animals ; Cardiomyopathy, Hypertrophic ; drug therapy ; metabolism ; Connexin 43 ; metabolism ; Disease Models, Animal ; Male ; Myocardium ; metabolism ; Rats ; Rats, Wistar

β-myosin heavy chain mutations are the most frequently identified basis for hypertrophic cardiomyopathy (HCM). A transgenic mouse model (αMHC(403)) has been extensively used to study various mechanistic aspects of HCM. There is general skepticism whether mouse and human disease features are similar. Herein we compare morphologic and functional characteristics, and disease evolution, in a transgenic mouse and a single family with a MHC mutation. Ten male αMHC(403) transgenic mice (at t-5 weeks, -12 weeks, and -24 weeks) and 10 HCM patients from the same family with a β-myosin heavy chain mutation were enrolled. Morphometric, conventional echocardiographic, tissue Doppler and strain analytic characteristics of transgenic mice and HCM patients were assessed. Ten male transgenic mice (αMHC(403)) were examined at ages -5 weeks, -12 weeks, and -24 weeks. In the transgenic mice, aging was associated with a significant increase in septal (0.59±0.06 vs. 0.64±0.05 vs. 0.69±0.11 mm, P<0.01) and anterior wall thickness (0.58±0.1 vs. 0.62±0.07 vs. 0.80±0.16 mm, P<0.001), which was coincident with a significant decrease in circumferential strain (-22%±4% vs. -20%±3% vs. -19%±3%, P=0.03), global longitudinal strain (-19%±3% vs. -17%±2% vs. -16%±3%, P=0.001) and E/A ratio (1.9±0.3 vs. 1.7±0.3 vs. 1.4±0.3, P=0.01). The HCM patients were classified into 1st generation (n=6; mean age 53±6 years), and 2nd generation (n=4; mean age 32±8 years). Septal thickness (2.2±0.9 vs. 1.4±0.1 cm, P<0.05), left atrial (LA) volume (62±16 vs. 41±5 mL, P=0.03), E/A ratio (0.77±0.21 vs. 1.1±0.1, P=0.01), E/e' ratio (25±10 vs. 12±2, P=0.03), global left ventricular (LV) strain (-14%±3% vs. -20%±3%, P=0.01) and global LV early diastolic strain rate (0.76±0.17 s(-1) vs. 1.3±0.2 s-1, P=0.01) were significantly worse in the older generation. In β-myosin heavy chain mutations, transgenic mice and humans have similar progression in morphologic and functional abnormalities. The αMHC(403) transgenic mouse model closely recapitulates human disease.
Adult ; Age Factors ; Aging ; Animals ; Cardiomyopathy, Hypertrophic, Familial ; genetics ; physiopathology ; Cross-Sectional Studies ; Disease Models, Animal ; Echocardiography, Doppler ; Female ; Heart ; physiopathology ; Humans ; Male ; Mice, Transgenic ; Middle Aged ; Myocardium ; metabolism ; pathology ; Myosin Heavy Chains ; genetics ; Phenotype ; Species Specificity ; Young Adult

OBJECTIVEMitochondrial disease is a group of energy metabolic disorders, characterized by involvement of multisystem with high energy requirements. Encephalomyopathies are common clinical findings of the mitochondrial diseases. However, mitochondrial cardiac damage is not rare. In this study, the clinical, biological, and genetic analyses were performed in three patients with mitochondrial cardiac damage, in order to understand the characteristics of mitochondrial diseases.

METHODThree girls presented with arrhythmia and cardiac enlargement from the age of 3, 4 and 8 years respectively. They were admitted into the Peking University First Hospital. Infection, autoimmune diseases, aminoacidopathies, organic acidurias, mitochondrial-fatty acid oxidation defects, and lysosomal storage disease were excluded by routine laboratory examinations and metabolic analysis for blood amino acids, acylcarnitines, urinary organic acids, and lysosome activity assay. Peripheral leukocytes mitochondrial respiratory chain enzyme I to V activities were measured by spectrophotometry. The entire sequence of the mitochondrial DNA was analyzed.

RESULTIn two patients (case 1 and case 3), hypertrophic cardiomyopathy and grade I to grade II of cardiac function were found. One patient (case 2) was diagnosed with arrhythmia and grade I of cardiac function. Increased creatine phosphokinase and creatine kinase isoenzyme MB were observed. Mitochondrial respiratory chain complex deficiencies were indentified in the three patients. Patient 1 had combined deficiencies of complex III and V. The activity of complex I+III was 18.7 nmol/(min·mg mitochondrial protein) (control 84.4 ± 28.5). The activity of complex V was 20.4 nmol/(min·mg mitochondrial protein) (control 103.7 ± 29.2). In her mitochondrial gene, A14693G on tRNA(Glu) and T16519C on D-loop were found. Patient 2 had an isolated complex I deficiency. The activity was 22.0 nmol/(min·mg mitochondrial protein) (control 44.0 ± 5.4). A16183C, T16189C and G15043A mutations on D-loop were found. Patient 3 had a combined deficiency of complex IV and V. The activity of complex IV was 21.0 nmol/(min·mg mitochondrial protein) (control 54.1 ± 12.3). The activity of complex V was 23.2 nmol/(min·mg mitochondrial protein) (control 103.7 ± 29.2). C253T and C16187T mutations on D-loop were detected. Haplotype analysis showed that three patients belong to H2a2a. Improvement was observed after the treatment with L-carnitine, coenzyme Q10, vitamin C and E. At present, the patients are 7, 5 and 8 years old. Although excise intolerance still persists, they had a good general condition with normal school life.

CONCLUSIONThe mitochondrial diseases with cardiac damage show cardiomyopathy, arrhythmia and exercise intolerance. Many kinds of mitochondrial respiratory chain deficiency were observed. A14693G in mitochondrial tRNA(Glu) gene is probably one of the causes in China.

Arrhythmias, Cardiac ; diagnosis ; genetics ; metabolism ; Biomarkers ; blood ; urine ; Cardiomyopathy, Hypertrophic ; diagnosis ; genetics ; metabolism ; Child ; Child, Preschool ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Electron Transport Chain Complex Proteins ; deficiency ; genetics ; metabolism ; Female ; Humans ; Male ; Mitochondria, Heart ; enzymology ; pathology ; Mitochondrial Diseases ; diagnosis ; genetics ; metabolism ; Mutation

OBJECTIVETo explore changes of mitochondrial structure and functions, as well as the protection of ligustrazine in the process of myocardial hypertrophy.

METHODSNeonatal myocardial cells were isolated and cultured with angiotensin II (Ang II) for 72 or 96 h. The total protein content was detected using BCA method. The cell diameter was measured by inverted microscope, by which to reflect the proliferation situation of cardiomyocytes. The mitochondrial membrane potential (MMP) was measured by fluorescence microscope. The mitochondrial monoamine oxidase (MAO) activity was detected by spectrophotometer. The mitochondrial cytochrome oxidase (COX) activity and the mitochondrial damage percentage were detected by microplate reader, by which to reflect the damage of mitochondrial outer membrane's structure and the membranes' function. Also, cells were treated with ligustrazine and losartan and then the pharmacological effects on the mitochondrial structure and functions in the myocardial cells treated with Ang II were observed.

RESULTSAt 72 h and 96 h, when compared with the blank group, cells treated with Ang II had increased total protein content (P < 0.01) and enlarged diameter (P < 0.01). Treated with Ang II, the MAO activity and the outer membrane damage percentage of myocardial cells significantly increased (P < 0.01), and mitochondrial COX activity and the mitochondrial MMP significantly decreased (P < 0.01). Compared with the model group at the same time period, ligustrazine significantly reduced myocardial cells' total protein content and myocardial cell diameter, and significantly decreased myocardial cells' MAO activity, increased mitochondrial COX activity, improved the outer membrane damage percentage and inner membrane MMP at 72 and 96 h, all showing statistical difference (P < 0.01, P < 0.05).

CONCLUSIONSDuring the process of myocardial hypertrophy existed the damage to the mitochondrial structure and functions. Ligustrazine protected the mitochondrial structure and functions of the myocardial cells in reversing Ang II induced myocardial cell hypertrophy.

Angiotensin II ; adverse effects ; Animals ; Cardiomyopathy, Hypertrophic ; chemically induced ; metabolism ; pathology ; Cells, Cultured ; Electron Transport Complex IV ; metabolism ; Mitochondria, Heart ; drug effects ; enzymology ; Monoamine Oxidase ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the changes of collagen fibers and the expression of osteopontin in the left ventricle in cases of hypertrophic cardiomyopathy (HCM), along with the significance of their potential forensic application. METHODS: Fifteen cases of HCM, 15 cases of coronary heart disease with cardiac hypertrophy and 20 cases of traffic accidents were selected as HCM group, coronary heart disease group and control group, respectively. Collagen volume fraction and osteopontin expression were observed and compared by HE staining, Masson trichrome staining and immunohistochemistry methods. Imaging and statistical methods were used for quantitative analysis. RESULTS: Collagen volume fraction in left ventricle of HCM and coronary heart disease were significantly higher than that in the control group (P < 0.05), which was not significantly different between the HCM group and the coronary heart disease group. The integral light density value of osteopontin in left ventricular cardiomyocytes of the HCM group and the coronary heart disease group were significantly higher than that of the control group (P< 0.05), and the value of the HCM group was also significantly higher than that of coronary heart disease group (P < 0.05). CONCLUSION The increased contents of collagen fibers and the overexpression of osteopontin may play an important role in myocardial fibrosis, and they can be used as markers in aid of diagnosing sudden death due to HCM.
Cardiomyopathy, Hypertrophic/physiopathology* ; Case-Control Studies ; Collagen/metabolism* ; Coronary Disease/physiopathology* ; Death, Sudden, Cardiac/etiology* ; Female ; Fibrosis ; Forensic Pathology ; Heart Ventricles/pathology* ; Humans ; Immunohistochemistry ; Male ; Myocardium/pathology* ; Osteopontin/metabolism* ; Staining and Labeling

OBJECTIVETo investigate the alteration of the adiponectin signal pathway in hypertrophic myocardium of spontaneous hypertensive rats (SHR) and to observe the effects of Gadol (GD) and Ganoderma spores (GS) on the hemodynamic parameters and the adiponectin signal pathway of SHR.

METHODSSHRs, 8 weeks old, were randomly divided into four groups: the untreated group, and the three treated groups treated with GD, GS, and GD + GS respectively by gastrogavage for 4 weeks. Controlled with 8-week-old WKY rats, the hemodynamic parameters in all rats were recorded through the carotid artery intubation; the serum level of adiponectin was determined with ELISA; the mRNA expressions of adiponectin receptors (AdipoRs) and carnitine palmitoyl transferase (CPT-1) were determined by RT-PCR; and the protein expression of adenosine monophosphate activated protein kinase (AMPK), both phosphorylated and un-phosphorylated, was detected by Western blot.

RESULTSCompared with the WKY rats, the systolic blood pressure (SBP), diastolic blood pressure (DBP) and myocardial hypertrophy index (MHI) in SHR were significantly higher; the serum levels of adiponectin and phosphorylated AMPK, mRNA expressions of AdipoR1 and CPT-1 in SHR heart tissue were lower (P < 0.05). Compared with the SHR, medication of GD and GS, either alone or in combination, could reduce SBP, DBP and MHI significantly (P < 0.01, P < 0.05), and elevate the mRNA expression of CPT-1 (P < 0.05) in heart, but levels of adiponectin, AdipoR1 and phosphorylated AMPK could only be raised by combined use of the two (P < 0.05).

CONCLUSIONSAdiponectin signal transduction pathway alteration presents in the myocardium of SHR, which might be one of the molecular mechanisms that cause hypertrophic metabolic abnormality. GD and GS could improve the hemodynamic index in SHR, and enhance the level of adiponectin and the expression of its related signal transduction molecules.

Adiponectin ; metabolism ; physiology ; Animals ; Cardiomyopathy, Hypertrophic ; etiology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Ganoderma ; chemistry ; Hemodynamics ; drug effects ; Hypertension ; complications ; Male ; Random Allocation ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Rhodiola ; chemistry ; Signal Transduction ; drug effects ; Spores

OBJECTIVETo explore the molecular biological mechanism for tanshinone II A reversing left ventricular hypertrophy, it would be studying the effect of tashinone on the endothelial nitric oxide synthase (eNOS) and protein kinase C (PKC) in the hypertrophic cadiocyte of rats suffered abdominal aorta constriction.

METHODSD rats were operated with abdominal aorta constriction and 8 rats were done with sham surgery. After 4 weeks, all rats were divided into 4 groups: myocardial hypertrophy group, low dose tanshinone II A group (10 mg x kg(-1) x d(-1)), high dose tanshinone II A group (20 mg x kg(-1) x d(-1)) and valsartan group (10 mg x kg(-1) d(-1) intragastric administration). 8 weeks later, the rats were used to measure the left ventricular mass index (LVMI) with the tissue of left ventricle and myocardial fiber dimension (MFD) by pathological section and HE stain, to detect the nitric oxide content by nitrate reductase, to detect the genic expression of eNOS by RT-PCR and to detect the activity of protein kinase C (PKC) by Western blotting.

RESULT1) The blood pressure in group myocardial hypertrophy [(186 +/- 13) mmHg] and tansginone II A [low and high dose (188 +/- 11,187 +/- 14) mmHg] was obviously higher than that in group sham surgery and valsartan group [vs (117 +/- 8, 136 +/- 15) mmHg, P < 0.01]. But there was no difference between group myocardial hypertrophy and group tanshinone II A (low and high dose). 2) The LVMI and MFD were obviously higher in group tanshinone II A low and high dose) and group valsartan than those in group sham surgery (P < 0.05), and lower than those in group myocardial hypertrophy (P < 0.01). 3) The NO level was obviously higher in group tanshinone II A (low and high dose) and group valsartan than that in group myocardial hypertrophy (12.78 +/- 1.66, 11.95 +/- 1.39, 12.26 +/- 2.08 vs 5.83 +/- 1.06) micromol x L(-1), (P < 0.01 ), and lower than that in group sham surgery (vs 19.35 +/- 1.47) micromol x L(-1), (P < 0.05). 4) The expressive level of eNOS mRNA and protein in myocardial hypertrophy group was less than that in other groups (P < 0.01). And valsartan group was less than tanshinone II A groups and sham surgery group (P < 0.05), but there were no difference among the two tanshinone II A groups and sham surgery group. 5) The level of PKC protein in group myocardial hypertrophy was obviously higher than that in all the other groups (1.291 +/- 0.117 vs 0.563 +/- 0.094, 0.605 +/- 0.051, 0.519 +/- 0.062, 0.827 +/- 0.086, P < 0.01), and the level in group valsartan was higher than that in group sham operation and group tanshinone II A (low and high dose).

CONCLUSIONNO/NOS system in local myocardium has close relationship with the pathological process for myocardial hypertrophy. Tanshinone II A can produce the pharmacological action to reverse myocardial hypertrophy by inhibiting the activity of PKC and promoting the genic expression of eNOS in local myocardium and the production of endogenous NO.

Animals ; Aorta, Abdominal ; pathology ; Benzofurans ; pharmacology ; Blood Pressure ; drug effects ; Cardiomyopathy, Hypertrophic ; complications ; enzymology ; physiopathology ; Constriction, Pathologic ; complications ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; drug effects ; enzymology ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Heart Ventricles ; drug effects ; metabolism ; pathology ; physiopathology ; Male ; Myocytes, Cardiac ; drug effects ; enzymology ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; genetics ; metabolism ; Protein Kinase C ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats

OBJECTIVETo explore the molecular biological mechanism of tanshinone II A (TSN) in preventing hypertensive left ventricular hypertrophy (HLVH) through studying the effects of TSN on angiotensin receptor (ATR) expression and free calcium ion ([Ca2+]i) in rats with hypertrophic myocardium caused by abdominal aorta constriction.

METHODSSD rats were established into HLVH model by abdominal aorta constriction operation, they were randomly divided into the model group, the three treated groups treated respectively with intra peritoneal injection of low dose TSN (10 mg/kg) and high dose TSN (20 mg/kg) and gastrogavage of Valsartan (10 mg/kg) once a day 4 weeks after modeling. Besides, 8 sham-operated SD rats were set up as the control group. Eight weeks later, rats' caudal arterial pressure was measured, and their hearts were taken for measuring the left ventricular mass index (LVMI) and myocardial fiber diameter (MFD) by HE stain of the pathological section. Moreover, the mRNA and protein expressions of AT1 and AT2 receptors in the left ventricular tissue were detected by RT-PCR and Western blot, and [Ca2+]i concentration was determined with laser-scanning confocal microscope.

RESULTS(1) The elevated blood pressure in the TSN treated groups, either high or low dose, remained unchanged, significantly higher than that in the control group and the Valsartan treated group (P < 0.01, P < 0.05). (2) LVMI and MFD in the three treated groups were significantly lower than those in the model group (P <0.01), respectively, although they were higher than those in the control group (P <0.05). (3) The mRNA and protein expressions of AT1 receptor were obviously lower in the three treated groups than those in the model groups (P < 0.05); but the lowering was more significant in the valsartan treated group (P < 0.05). (4) The mRNA and protein expressions of AT2 receptor were significantly higher in the Valsartan treated group as compared with other groups (P < 0.05), while the difference among the other groups showed no statistical significance (P > 0.05). (5) The elevated (Ca2+]i concentration in hypertrophic myocardium after modeling was significantly lowered after treatment in the three treated groups (P < 0.05), but the lowering in the high TSN treated group was more significant than that in the Valsartan treated group (P <0.05).

CONCLUSIONThe inhibition of TSN on myocardial hypertrophy is blood pressure independent, its mechanism is possibly related with the inhibition on AT1R gene expression and the blocking of free calcium ion influx in cardiac muscle cells. AT2 receptor may participate the effect of Valsartan in lowering blood pressure and reversing myocardial hypertrophy.

Animals ; Blood Pressure ; drug effects ; Calcium ; metabolism ; Cardiomyopathy, Hypertrophic ; drug therapy ; metabolism ; physiopathology ; Disease Models, Animal ; Diterpenes, Abietane ; Female ; Humans ; Male ; Myocardium ; metabolism ; Phenanthrenes ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Angiotensin ; metabolism