Targeted delivery of a proangiogenic and promyogenic protein for regeneration of diabetic ischemic limbs

靶向递送促血管生成和促肌生成蛋白以促进糖尿病缺血肢体的再生

• 批准号: 10616819
• 负责人: Jianjun Guan
• 金额: $ 58.25万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-02 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10616819
• 关键词: Acceleration; Blood Platelets; Cell Survival; Cell membrane; Cell physiology; Cellular Metabolic Process; Clinical; Diabetic mouse; Diameter; Diffusion; Encapsulated; Endothelial Cells; Endothelium; Engineering; Environment; Exhibits; Fibrosis; Glucose; Growth Factor; Hyperglycemia; Impairment; Inferior; Inflammation; Injections; Ischemia; Limb Salvage; Limb structure; Membrane; Modeling; Morphogenesis; Mus; Muscle Cells; Muscle Fibers; Muscle function; Myoblasts; Natural regeneration; Peptides; Phosphotransferases; Principal Investigator; Proliferating; Property; Protein Engineering; Proteins; Reporting; Signal Pathway; Skeletal Muscle; Testing; Therapeutic Effect; Time; Tissues; Treatment Efficacy; Tube; Vascularization; angiogenesis; blood perfusion; cell motility; controlled release; critical limb Ischemia; density; diabetic; diabetic patient; effective therapy; efficacy testing; functional improvement; improved; innovation; limb amputation; limb ischemia; migration; mortality; myogenesis; nanoparticle; novel; novel strategies; preservation; programs; repaired; satellite cell; targeted delivery

Program Director/Principal Investigator (Last, First, Middle): GUAN, JIANJUN Project Summary Diabetic patients with critical limb ischemia (CLI) have significantly high rates of limb amputation and mortality. CLI is featured by extremely low blood perfusion and degenerated skeletal muscle. Accordingly, regeneration of vasculature and skeletal muscles will salvage the limbs. Yet the poor endothelial and skeletal muscle cell survival, and inferior cell functions under the hyperglycemia and ischemic conditions of diabetic CLI impair the limb repair. Currently, there is no effective treatment available although growth factor therapy represents a promising strategy. However, growth factor therapy has relatively low therapeutic efficacy in regenerating both vasculature and skeletal muscles, as multiple growth factors are simultaneously needed for vascularization and myogenesis, and these cannot be readily delivered by current approaches. In this project, we propose to use a novel TRIM72 protein with both pro-angiogenic and pro-myogenic properties to regenerate vasculature and skeletal muscles in diabetic CLI. The TRIM72 will be engineered to have longer retention time (slower diffusion rate) in ischemic tissue, thus exhibiting longer therapeutic effect. To deliver the engineered TRIM72 (ETRIM72), it will be encapsulated into ischemic limb-targeting nanoparticles, followed by delivering via clinically attractive IV injection. The nanoparticles will then predominantly accumulate in the ischemic limbs and gradually release ETRIM72. The released protein will promote vascularization and myogenesis by (1) improving the survival of endothelial cell and skeletal muscle cell through cell membrane repair, and activation of cell survival kinase; and (2) stimulating endothelial cell and skeletal muscle cell migration and morphogenesis under the hyperglycemia and ischemic conditions of diabetic CLI. In our preliminary studies, we have developed ETRIM72 by genetically fusing TRIM72 with peptide CSTSMLKAC that targets ischemic environment of ischemic limbs. This first version of ETRIM72 was able to retain in the ischemic limbs significantly longer than TRIM72. After IV injection of ischemic limb-targeting, ETRIM72-releasing nanoparticles, the released ETRIM72 significantly promoted regeneration of both vasculature and skeletal muscles in diabetic ischemic limbs. The function of TRIM72 in promoting vascularization and myogenesis under hyperglycemia and ischemic conditions has not been reported before. Based on our preliminary studies, we hypothesize that controlled release of ETRIM72 will simultaneously increase endothelial and skeletal muscle cell survival, migration and morphogenesis under hyperglycemia and ischemic conditions, leading to accelerated regeneration of both vasculature and skeletal muscles in diabetic ischemic limbs. Aim #1 will test the hypothesis that optimal ETRIM72 release profiles will significantly promote survival, migration and morphogenesis of endothelial cells and myoblasts under high glucose and ischemic conditions. Aim #2 will test efficacy of the ETRIM72-releasing nanoparticles using diabetic murine limb ischemia model. This project is innovative because it engineers a novel proangiogenic and promyogenic protein to simultaneously regenerate vasculature and skeletal muscles in diabetic ischemic limbs. The longer tissue retention time of the engineered protein, together with localized and controlled release are expected to significantly improve therapeutic efficacy. OMB No. 0925-0001/0002 (Rev. 03/2020 Approved Through 02/28/2023) Page Continuation Format Page

项目主任/首席研究员（后、一、中）：管建军 项目概要 患有严重肢体缺血（CLI）的糖尿病患者肢体截肢率和死亡率显着升高。 CLI 的特点是血液灌注极低和骨骼肌退化。因此，再生 脉管系统和骨骼肌将挽救四肢。然而内皮细胞和骨骼肌细胞的存活率较差， 糖尿病 CLI 的高血糖和缺血条件下的劣质细胞功能会损害肢体修复。 目前，尽管生长因子疗法是一种有前途的策略，但尚无有效的治疗方法。 然而，生长因子疗法在血管和骨骼再生方面的治疗效果相对较低。 肌肉，因为血管形成和肌肉生成同时需要多种生长因子，而这些不能 可以通过当前的方法轻松实现。 在这个项目中，我们建议使用一种具有促血管生成和促肌生成特性的新型 TRIM72 蛋白 再生糖尿病性 CLI 中的脉管系统和骨骼肌。 TRIM72 将被设计为具有更长的 在缺血组织中的保留时间（较慢的扩散速率），从而表现出更长的治疗效果。为了交付 工程化的 TRIM72 (ETRIM72)，它将被封装到缺血肢体靶向纳米颗粒中，然后 通过临床上有吸引力的静脉注射进行输送。然后纳米颗粒将主要积聚在缺血区域 四肢并逐渐释放ETRIM72。释放的蛋白质将通过 (1) 促进血管形成和肌生成 通过细胞膜修复和细胞活化提高内皮细胞和骨骼肌细胞的存活率 生存激酶； (2)刺激内皮细胞和骨骼肌细胞迁移和形态发生 糖尿病 CLI 的高血糖和缺血状况。 在我们的初步研究中，我们通过将TRIM72与肽CSTSMLKAC进行基因融合来开发ETRIM72 针对缺血肢体的缺血环境。 ETRIM72 的第一个版本能够保留在缺血 四肢明显长于 TRIM72。静脉注射靶向缺血肢体、释放 ETRIM72 的纳米颗粒后， 释放的 ETRIM72 显着促进糖尿病患者血管和骨骼肌的再生 四肢缺血。 TRIM72在高血糖和肌生成中促进血管化和肌生成的功能 以前没有报道过缺血性疾病。 根据我们的初步研究，我们假设 ETRIM72 的控释会同时增加 高血糖和缺血条件下内皮和骨骼肌细胞的存活、迁移和形态发生 导致糖尿病缺血肢体的脉管系统和骨骼肌加速再生。 目标 #1 将测试最佳 ETRIM72 释放曲线将显着促进生存和迁移的假设 高葡萄糖和缺血条件下内皮细胞和成肌细胞的形态发生。目标 #2 将进行测试 使用糖尿病小鼠肢体缺血模型研究 ETRIM72 释放纳米颗粒的功效。 该项目具有创新性，因为它设计了一种新型的促血管生成和促肌生成蛋白 同时再生糖尿病缺血肢体的脉管系统和骨骼肌。组织保留时间越长 工程蛋白的时间以及局部和受控释放预计将显着改善 治疗功效。 OMB 编号 0925-0001/0002（修订版 03/2020 已批准至 02/28/2023） 页面延续格式页面