Glutaminase in Arterial Injury and Disease

谷氨酰胺酶在动脉损伤和疾病中的作用

• 批准号: 10473678
• 负责人: WILLIAM DURANTE
• 金额: $ 39.07万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-23 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10473678
• 关键词: Acid-Base Equilibrium; Americas; Ammonia; Animals; Apoptosis; Arterial Injury; Arteries; Atherosclerosis; Attenuated; Biological Models; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Carotid Arteries; Cell Migration Inhibition function; Cell Proliferation; Cell physiology; Collagen; DNA biosynthesis; Data; Diabetes Mellitus; Diabetic mouse; Disease; Enzymes; Extracellular Signal Regulated Kinases; Fibrosis; Gene Delivery; Gene Transfer; Generations; Genetic; Glucose; Glutamates; Glutaminase; Glutamine; Goals; Growth Factor; Hypertension; In Vitro; Insulin-Dependent Diabetes Mellitus; Laboratories; Lesion; Liver Fibrosis; Mediating; Mediator of activation protein; Metabolism; Mitochondria; Non-Insulin-Dependent Diabetes Mellitus; Pharmacology; Play; Process; Production; Proline; Protein Isoforms; Pulmonary Hypertension; Regulation; Risk; Rodent; Role; Seminal; Signal Transduction; Smooth Muscle Myocytes; Stroke; Testing; Time; Transcription Factor AP-1; Transcriptional Activation; United States; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; adenoviral-mediated; angiogenesis; arterial remodeling; arterial stiffness; base; cell motility; cost; diabetic; diabetic patient; experimental study; heme oxygenase-1; in vivo Model; injured; migration; mortality; mouse model; neointima formation; neurotransmission; new therapeutic target; novel; overexpression; patient population; percutaneous coronary intervention; prevent; response; response to injury; restenosis; success; tumor growth; vascular abnormality; vascular smooth muscle cell proliferation

PROJECT SUMMARY The broad long-term goal of this proposal is to establish the enzyme glutaminase-1 (GLS1) as a critical mediator of lesion formation in arterial injury and disease. GLS1 is a mitochondrial enzyme that metabolizes glutamine (Gln) to glutamate (Glu) and ammonia (NH3). GLS1 plays a critical role in neurotransmission, acid- base balance, angiogenesis, hepatic fibrosis, and tumor growth, but little is known regarding the function of GLS1 in vascular smooth muscle cells (SMCs). However, preliminary studies demonstrated that the proliferation, migration, and survival of SMCs is highly dependent on the presence of Gln. It was also discovered that SMCs exclusively express the GLS1 isoform, that overexpression of GLS1 stimulates SMC migration, and that inhibition of GLS1 activity or expression blocks SMC DNA synthesis or migration, respectively. Moreover, Glu, but not NH3, substitutes for Gln in promoting SMC proliferation and collagen synthesis. Additional experiments revealed that growth factors and glucose stimulate GLS1 activity in vascular SMCs. Final pilot experiments also demonstrated that arterial injury and diabetes induces the expression of GLS1 in the vessel wall, and that inhibition of GLS1 activity or genetic depletion of GLS1 attenuates neointima formation following arterial injury as well as arterial fibrosis and stiffening in diabetic animals. Based on these findings, it is proposed that GLS1 plays an integral role in aberrant arterial remodeling by stimulating SMC proliferation, migration, collagen synthesis, and survival by metabolizing Gln to Glu and NH3. This hypothesis will be tested in three interrelated specific aims. In aim 1, the role of GLS1 in regulating SMC function will be determined. These studies will investigate the effect of GLS1 gene delivery on SMC proliferation, migration, collagen synthesis and survival, and determine the role of the various GLS1 products on these processes. They will also explore if the induction of GLS1 by growth factors contributes to their ability to promote SMC proliferation, migration, and collagen synthesis. In aim 2, the role of GLS1 in regulating the arterial response to injury will be established. These studies will examine the time-course of GLS1 expression in injured rodent carotid arteries, and determine if pharmacological inhibition of GLS1 activity, silencing GLS1 expression, or genetic deletion of GLS1 in SMCs attenuates the remodeling response following arterial injury. In aim 3, the role of GLS1 in aberrant arterial remodeling and hypertension in diabetes will be investigated. These studies will examine the effect of glucose on GLS1 expression both in cultured vascular SMCs and in arteries from diabetic mice. In addition, they will examine if GLS1 contributes to glucose-mediated alterations in SMC function, arterial remodeling, and hypertension in diabetic mice. It is anticipated that these studies will establish GLS1 as a key regulator of SMC proliferation, migration, collagen synthesis, and survival. They may also also identify GLS1 and its products as novel contributors to lesion formation following arterial injury, and establish GLS1 as a new translational target in treating abnormal remodeling and hypertension in diabetes.

项目概要 该提案的广泛长期目标是将谷氨酰胺酶-1 (GLS1) 确立为关键的酶 动脉损伤和疾病中病变形成的介质。 GLS1 是一种线粒体酶，负责代谢 谷氨酰胺 (Gln) 转化为谷氨酸 (Glu) 和氨 (NH3)。 GLS1 在神经传递、酸- 基础平衡、血管生成、肝纤维化和肿瘤生长，但对其功能知之甚少。 血管平滑肌细胞 (SMC) 中的 GLS1。然而，初步研究表明， SMC 的增殖、迁移和存活高度依赖于 Gln 的存在。也曾是 发现 SMC 只表达 GLS1 同工型，GLS1 的过度表达会刺激 SMC 迁移，并且抑制 GLS1 活性或表达会阻断 SMC DNA 合成或迁移， 分别。此外，Glu（而非 NH3）可替代 Gln 促进 SMC 增殖和胶原蛋白 合成。其他实验表明，生长因子和葡萄糖会刺激血管中的 GLS1 活性。 SMC。最终的初步实验还表明，动脉损伤和糖尿病会诱导 血管壁中的 GLS1，抑制 GLS1 活性或 GLS1 基因缺失会减弱新内膜 糖尿病动物动脉损伤后的形成以及动脉纤维化和硬化。基于这些 研究结果表明，GLS1 通过刺激 SMC 在异常动脉重塑中发挥着不可或缺的作用 通过将 Gln 代谢为 Glu 和 NH3 来促进增殖、迁移、胶原蛋白合成和存活。这个假设 将在三个相互关联的具体目标中进行测试。在目标 1 中，GLS1 在调节 SMC 功能中的作用将是 决定。这些研究将调查 GLS1 基因传递对 SMC 增殖、迁移、 胶原蛋白的合成和存活，并确定各种 GLS1 产品在这些过程中的作用。 他们还将探讨生长因子诱导 GLS1 是否有助于促进 SMC 增殖、迁移和胶原蛋白合成。在目标 2 中，GLS1 在调节动脉反应中的作用 伤害将成立。这些研究将检查受伤啮齿动物中 GLS1 表达的时间过程 颈动脉，并确定是否通过药物抑制 GLS1 活性、沉默 GLS1 表达或 SMC 中 GLS1 的基因缺失会减弱动脉损伤后的重塑反应。在目标 3 中， 将研究 GLS1 在异常动脉重塑和糖尿病高血压中的作用。这些研究 将检查葡萄糖对培养的血管 SMC 和动脉中 GLS1 表达的影响 糖尿病小鼠。此外，他们还将检查 GLS1 是否有助于葡萄糖介导的 SMC 改变 糖尿病小鼠的功能、动脉重塑和高血压。预计这些研究将 确立 GLS1 作为 SMC 增殖、迁移、胶原蛋白合成和存活的关键调节因子。他们可能 还确定 GLS1 及其产物是动脉损伤后病变形成的新贡献者，并且 将 GLS1 确立为治疗糖尿病异常重塑和高血压的新转化靶点。