Cyclic nucleotide phosphodiesterase regulation in vascular calcification

血管钙化中环核苷酸磷酸二酯酶的调节

• 批准号: 10641917
• 负责人: Yujun Cai
• 金额: $ 62.97万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641917
• 关键词: Address; Amputation; Arterial Medias; Arteries; Attenuated; Bioinformatics; Blood Vessels; Cardiovascular system; Cell physiology; Cells; Chronic Kidney Failure; Cyclic AMP; Cyclic GMP; Cyclic Nucleotides; Deposition; Diabetes Mellitus; Drug Targeting; Durapatite; Enzymes; Event; Extracellular Matrix; Extracellular Matrix Degradation; Family; Family member; Genes; Goals; Human; In Vitro; Ischemia; Isoenzymes; Knockout Mice; Leg; Lower Extremity; MAP Kinase Gene; Matrix Metalloproteinases; Medial; Mediating; Metalloproteases; Morbidity - disease rate; Pathologic; Patients; Peripheral arterial disease; Pharmaceutical Preparations; Physiological; Play; Process; Protein Isoforms; Rattus; Regulation; Risk; Rodent Model; Role; Second Messenger Systems; Series; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Stromelysin 1; Therapeutic; Tibial Arteries; Time; Up-Regulation; Vascular Smooth Muscle; Vascular calcification; calcification; human disease; improved outcome; in vivo; inhibitor; inorganic phosphate; insight; knock-down; loss of function; molecular targeted therapies; mortality; mouse model; novel; novel therapeutic intervention; osteogenic; overexpression; p38 Mitogen Activated Protein Kinase; phosphoric diester hydrolase; prevent; therapy development

PROJECT SUMMARY Arterial calcification results from the deposition of calcium hydroxyapatite crystals in the vessel wall. It is highly prevalent in patients with chronic kidney disease (CKD) and diabetes as well as those with peripheral artery disease (PAD). When located in the arterial media, calcification is strongly associated with increased cardiovascular morbidity and mortality. Calcification is a highly regulated process controlled by a series of endogenous stimulators and inhibitors. Elevated phosphate levels can induce osteogenic transformation of vascular smooth muscle cells (SMCs). These cells release extracellular matrix-degrading enzymes including the matrix metalloproteinases (MMPs) that contribute to calcification. Despite significant progress, arterial calcification continues to be poorly understood and no useable drugs to prevent or treat it have been developed. For this reason, the process of identifying novel molecular targets, and developing therapies to target them are critical for improving outcomes in our vascular patients. The second messenger cyclic nucleotides cAMP and cGMP play important regulatory roles in a variety of human diseases that are controlled by distinct cyclic nucleotide PDE isozymes that have proven to be ideal and feasible drug targets for the treatment of human diseases. The function and regulation of PDEs in arterial calcification, however, remains unknown. This proposal is based on our preliminary data showing that phosphodiesterase 1 0A (PDE10A) is the most highly induced isoform among all PDE genes in a rodent calcification model. PDE10A is also markedly increased in calcifying VSMCs in vitro, calcified arteries in vivo, and calcified human tibial arteries from patients with PAD. Knockdown and inhibition of PDE10A significantly attenuate phosphate-induced VSMC osteogenic transformation and calcification in vitro, and deficiency of PDE10A reduces arterial calcification in vivo. Using bioinformatics analyses and a loss-of-function strategy, we have shown that the MMP family member 3 (MMP- 3, stromelysin-1) can be regulated by PDE10A in VSMCs cultured in a calcification medium. Our further preliminary mechanistic results have shown that both knockdown and inhibition of PDE1 0A block p38 MAPK activation in VSMCs during calcification. We have additionally found that inhibition of p38 MAPK attenuates MMP-3 upregulation under calcifying conditions. In this project, we propose that PDE10A mediates arterial calcification by regulating p38 MAPK-MMP-3 signaling. In this series of experiments, we will establish the role of PDE10A in arterial calcification and provide insights into the potential use of PDE10A inhibition strategies to reduce calcification in patients with CKD and PAD. Our aims are to 1) investigate the actions of PDE10A in vascular SMC osteogenic transformation and arterial calcification, 2) examine the therapeutic potential of PDE10A inhibition in arterial calcification, and 3) assess the significance of p38-MAPK-MMP-3 signaling in PDE10A-mediated vascular calcification.

项目概要 动脉钙化是羟基磷灰石钙晶体在血管壁沉积的结果。它是高度 常见于慢性肾病 (CKD) 和糖尿病患者以及外周动脉患者 疾病（PAD）。当位于动脉中膜时，钙化与钙化的增加密切相关。 心血管发病率和死亡率。钙化是一个高度调控的过程，受一系列因素控制 内源性刺激剂和抑制剂。磷酸盐水平升高可诱导成骨转化 血管平滑肌细胞（SMC）。这些细胞释放细胞外基质降解酶，包括 有助于钙化的基质金属蛋白酶（MMP）。尽管取得了重大进展，但动脉 人们对钙化仍然知之甚少，并且尚未开发出可用于预防或治疗钙化的药物。 因此，识别新的分子靶标并开发针对它们的疗法的过程是 对于改善血管患者的预后至关重要。第二信使环核苷酸cAMP和 cGMP 在受不同循环控制的多种人类疾病中发挥着重要的调节作用 核苷酸PDE同工酶已被证明是治疗人类理想且可行的药物靶标 疾病。然而，PDE 在动脉钙化中的功能和调节仍然未知。这 该提案基于我们的初步数据，显示磷酸二酯酶 1 0A (PDE10A) 是最高度 在啮齿动物钙化模型中所有 PDE 基因中诱导同种型。 PDE10A 也显着增加 体外钙化 VSMC、体内钙化动脉以及来自 PAD 患者的钙化人胫动脉。 PDE10A 的敲低和抑制显着减弱磷酸盐诱导的 VSMC 成骨 体外的转化和钙化，PDE10A 的缺乏会减少体内的动脉钙化。使用 通过生物信息学分析和功能丧失策略，我们发现 MMP 家族成员 3 (MMP- 3、stromelysin-1) 可以在钙化培养基中培养的 VSMC 中受到 PDE10A 的调节。我们进一步 初步机制结果表明，PDE1 0A 的敲低和抑制均可阻断 p38 MAPK 钙化过程中 VSMC 的激活。我们还发现 p38 MAPK 的抑制会减弱 钙化条件下 MMP-3 上调。在这个项目中，我们建议 PDE10A 介导动脉 通过调节 p38 MAPK-MMP-3 信号传导来抑制钙化。在这一系列的实验中，我们将建立角色 PDE10A 在动脉钙化中的作用，并提供有关 PDE10A 抑制策略潜在用途的见解 减少 CKD 和 PAD 患者的钙化。我们的目标是 1) 研究 PDE10A 在 血管SMC成骨转化和动脉钙化，2）检查治疗潜力 动脉钙化中的 PDE10A 抑制，以及 3) 评估 p38-MAPK-MMP-3 信号传导在动脉钙化中的重要性 PDE10A 介导的血管钙化。