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: 10521675
• 负责人: Yujun Cai
• 金额: $ 65.31万
• 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/10521675
• 关键词: Address; Amputation; Arterial Medias; Arteries; Attenuated; Bioinformatics; Blood Vessels; Cardiovascular system; Cell physiology; Cells; Chronic Kidney Failure; Crystallization; 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; 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.

项目摘要 动脉钙化是由羟基磷灰石晶体在血管壁中的沉积而产生的。它在患有慢性肾脏疾病（CKD）和糖尿病的患者以及外周动脉疾病（PAD）的患者中非常普遍。当位于动脉介质中时，钙化与心血管发病率和死亡率的增加密切相关。钙化是由一系列内源刺激剂和抑制剂控制的高度调节过程。磷酸盐水平升高会诱导血管平滑肌细胞（SMC）的成骨转化。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，但动脉钙化仍被了解不多，并且没有可用的药物来预防或治疗。因此，鉴定新的分子靶标的过程以及开发靶向目标的疗法对于改善血管患者的预后至关重要。第二信使环核苷酸营地和CGMP在多种人类疾病中起着重要的调节作用，这些人类疾病受到不同环状核苷酸PDE同工酶的控制，这些同性恋者已被证明是理想且可行的药物靶标，用于治疗人类疾病。然而，PDE在动脉钙化中的功能和调节仍然未知。该建议基于我们的初步数据，表明磷酸二酯酶1 0a（PDE10A）是啮齿动物钙化模型中所有PDE基因中最高度诱导的同工型。 PDE10A在体外钙化VSMC中也明显增加，体内钙化动脉，并从PAD患者中钙化人胫骨动脉。 PDE10A的敲低和抑制显着减弱了磷酸盐诱导的VSMC成骨转化和体外钙化，PDE10A的缺乏降低了体内动脉钙化。使用生物信息学分析和功能丧失策略，我们表明MMP家族成员3（MMP-3，Stromelysin-1）可以通过PDE10A在钙化培养基中培养的VSMC中调节。我们进一步的初步机械结果表明，钙化过程中VSMC中PDE1 0A块P38 MAPK激活的敲低和抑制。我们还发现，抑制p38 MAPK在钙化条件下会减弱MMP-3上调。在这个项目中，我们建议PDE10A通过调节p38 MAPK-MMP-3信号传导来介导动脉钙化。在这一系列实验中，我们将确定PDE10A在动脉钙化中的作用，并提供有关PDE10A抑制策略潜在使用以减少CKD和PAD患者钙化的潜在使用。我们的目的是1）研究PDE10A在血管成生成转化和动脉钙化中的作用，2）检查PDE10A抑制在动脉钙化中的治疗潜力，3）评估P38-MAPK-MMP-3信号在PDE10A中的显着性 - 介导的血管钙化。