Dominant role of CACNA1D exon mutations for blood pressure regulation

Background: CACNA1D gene, which encodes the α1 subunit of the Cav1.3 L-type calcium channel effectively regulates intracellular Ca2+ stability. In recent years, clinical studies have shown that the CACNA1D polymorphisms were associated with hypertension. Objective: The purpose of this study was to evaluate the effects of CACNA1D exon mutation on blood pressure (BP) in Sprague–Dawley rats. Methods: The rats with CACNA1D p.D307G, CACNA1D p.V936I or CACNA1D p.R1516Q were constructed using CRISPR-Cas9 technology. SBP measurements of rats were taken for 32 weeks. Tissue morphology of rats and vasoactive substances in serum was tested. Furthermore, the effects of L-type calcium channel blocker isradipine and endothelin-1 (ET-1) inhibitor BQ-123 on BP of double mutation rats (CACNA1D p.D307G/p.R1516Q) were tested. Then we examined the effects of CACNA1D gene mutation on gene expression in human umbilical vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs). Results: Elevated SBP and increased circulating ET-1 was observed in CACNA1D p.D307G mutant rats. Morphological assessments showed that the vascular, cardiac and renal remodeling could also be observed in rats with p.D307G mutant. Cav1.3 protein expression and calcineurin phosphatase activity in VSMCs of rats with CACNA1D p.D307G were increased in vitro, and the vascular ring tension test of mesenteric grade 3 arteries in CACNA1D p.D307G rats were increased in vivo. Furthermore, ET-1 expression were increased in isolated primary aortic endothelial cells in p.D307G mutant rats and transfected p.D307G mutant HUVECs. Finally, double heterozygosity rats with CACNA1D p.D307G/p.R1516Q or CACNA1D p.D307G/p.V936I further accelerated the rise of SBP compared with p.D307G mutation rats, and isradipine and BQ-123 reduced BP to the same extent in CACNA1D p.D307G/p.R1516Q rats. Conclusion: CACNA1D gene is key players in the regulation of blood pressure. CACNA1D mutation rat may be a new hypertension animal model.

[1]  C. Gomez-Sanchez,et al.  The landscape of molecular mechanism for aldosterone production in aldosterone-producing adenoma. , 2020, Endocrine journal.

[2]  S. Tomlins,et al.  Prevalence of somatic mutations in aldosterone-producing adenomas in Japanese. , 2020, The Journal of clinical endocrinology and metabolism.

[3]  Qifu Li,et al.  Genetic screening for monogenic hypertension in hypertensive individuals in a clinical setting , 2020, Journal of Medical Genetics.

[4]  M. Thuzar,et al.  Biochemical, Histopathological and Genetic Characterization of Posture Responsive and Unresponsive APAs. , 2020, The Journal of clinical endocrinology and metabolism.

[5]  S. Tomlins,et al.  Histological characterization of aldosterone-producing adrenocortical adenomas with different somatic mutations. , 2019, The Journal of clinical endocrinology and metabolism.

[6]  Luoping Zhang,et al.  Vasodilatory effect of formaldehyde via the NO/cGMP pathway and the regulation of expression of KATP, BKCa and L-type Ca2+ channels. , 2019, Toxicology letters.

[7]  Jun Ma,et al.  Combined effects of the rs9810888 polymorphism in calcium voltage-gated channel subunit alpha1 D (CACNA1D) and lifestyle behaviors on blood pressure level among Chinese children , 2019, PloS one.

[8]  Ji-Guang Wang,et al.  Genome-Wide Association Studies of Hypertension and Several Other Cardiovascular Diseases , 2019, Pulse.

[9]  Jie Dong,et al.  5-HTT, BMPR2, EDN1, ENG, KCNA5 gene polymorphisms and susceptibility to pulmonary arterial hypertension: A meta-analysis. , 2019, Gene.

[10]  Hui Chen,et al.  GW29-e0750 The CACNA1D exon mutations are possible novel risk locus for preeclampsia , 2018, Journal of the American College of Cardiology.

[11]  Giovanni Cugliari,et al.  Advances in the Genetics of Hypertension: The Effect of Rare Variants , 2018, International journal of molecular sciences.

[12]  U. Scholl.  Unanswered Questions in the Genetic Basis of Primary Aldosteronism , 2017, Hormone and Metabolic Research.

[13]  Hui Chen,et al.  GW28-e0620 Late diagnosis of Severe Delayed Postpartum Preeclampsia with a Rare CACNA1D Mutation , 2017 .

[14]  Xiaofeng Yu,et al.  Ginsenoside Rg3 induces ginsenoside Rb1-comparable cardioprotective effects independent of reducing blood pressure in spontaneously hypertensive rats , 2017, Experimental and therapeutic medicine.

[15]  Jing Xiao,et al.  Endothelin-receptor antagonist can reduce blood pressure in patients with hypertension: a meta-analysis , 2017, Blood pressure.

[16]  Haiqing Gao,et al.  Alleviative effect of grape seed proanthocyanidin extract on small artery vascular remodeling in spontaneous hypertensive rats via inhibition of collagen hyperplasia. , 2017, Molecular medicine reports.

[17]  F. Gao,et al.  Tetrahydroxystilbene Glucoside Inhibits Excessive Autophagy and Improves Microvascular Endothelial Dysfunction in Prehypertensive Spontaneously Hypertensive Rats. , 2016, The American journal of Chinese medicine.

[18]  Alexandra Pinggera,et al.  Pharmacology of L-type Calcium Channels: Novel Drugs for Old Targets? , 2015, Current molecular pharmacology.

[19]  Xueli Yang,et al.  Genome-wide association study in Chinese identifies novel loci for blood pressure and hypertension. , 2015, Human molecular genetics.

[20]  Z. Meng,et al.  Effects of gaseous sulfur dioxide and its derivatives on the expression of KATP, BKCa and L-Ca(2+) channels in rat aortas in vitro. , 2014, European journal of pharmacology.

[21]  Z. Meng,et al.  Effects of sodium metabisulfite on the expression of BK(Ca), K(ATP), and L-Ca(2+) channels in rat aortas in vivo and in vitro. , 2014, Journal of hazardous materials.

[22]  G. D. De Meyer,et al.  L-type Ca2+ channel blockers inhibit the window contraction of mouse aorta segments with high affinity. , 2014, European journal of pharmacology.

[23]  Annabelle L. Fonseca,et al.  Somatic and germline CACNA1D calcium channel mutations in aldosterone-producing adenomas and primary aldosteronism , 2013, Nature Genetics.

[24]  A. Saiki,et al.  Protective effects of efonidipine, a T- and L-type calcium channel blocker, on renal function and arterial stiffness in type 2 diabetic patients with hypertension and nephropathy. , 2009, Journal of atherosclerosis and thrombosis.

[25]  T. Miyata,et al.  Genetic polymorphisms of L-type calcium channel alpha1C and alpha1D subunit genes are associated with sensitivity to the antihypertensive effects of L-type dihydropyridine calcium-channel blockers. , 2009, Circulation journal : official journal of the Japanese Circulation Society.

[26]  C. Bronner,et al.  Angiotensin II-Induced Hypertension Is Associated with a Selective Inhibition of Endothelium-Derived Hyperpolarizing Factor-Mediated Responses in the Rat Mesenteric Artery , 2009, Journal of Pharmacology and Experimental Therapeutics.

[27]  M. Iglarz,et al.  Mechanisms of ET-1-induced Endothelial Dysfunction , 2007, Journal of cardiovascular pharmacology.

[28]  J. Pande,et al.  Aortic smooth muscle and endothelial plasma membrane Ca2+ pump isoforms are inhibited differently by the extracellular inhibitor caloxin 1b1. , 2006, American journal of physiology. Cell physiology.

[29]  Mario Fritsch Neves,et al.  Endothelium-Restricted Overexpression of Human Endothelin-1 Causes Vascular Remodeling and Endothelial Dysfunction , 2004, Circulation.

[30]  N. Klugbauer,et al.  Dominant role of smooth muscle L‐type calcium channel Cav1.2 for blood pressure regulation , 2003, The EMBO journal.

[31]  M. Yakubu,et al.  L-type voltage-dependent Ca2+ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis. , 2002, American journal of physiology. Cell physiology.

[32]  J. Pearson,et al.  Fibroblast matrix gene expression and connective tissue remodeling: role of endothelin-1. , 2001, The Journal of investigative dermatology.

[33]  D. Arveiler,et al.  The Lys198Asn polymorphism in the endothelin-1 gene is associated with blood pressure in overweight people. , 1999, Hypertension.

[34]  H. S. Kim,et al.  Genetic control of blood pressure and the angiotensinogen locus. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[35]  M. Yanagisawa,et al.  ECE-1: A membrane-bound metalloprotease that catalyzes the proteolytic activation of big endothelin-1 , 1994, Cell.

[36]  K. Fuxe,et al.  High blood pressure in transgenic mice carrying the rat angiotensinogen gene. , 1992, The EMBO journal.

[37]  M. Masu,et al.  Generation of transgenic mice with elevated blood pressure by introduction of the rat renin and angiotensinogen genes. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[38]  Y. Hirata,et al.  Endothelin is a potent mitogen for rat vascular smooth muscle cells. , 1989, Atherosclerosis.

[39]  L. Lerman,et al.  Peripheral vascular atherosclerosis in a novel PCSK9 gain‐of‐function mutant Ossabaw miniature pig model , 2018, Translational research : the journal of laboratory and clinical medicine.

[40]  D. Kohan,et al.  Regulation of blood pressure and salt homeostasis by endothelin. , 2011, Physiological reviews.