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

基本信息

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
项目类别：
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

项目摘要

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

项目总监/首席研究员（最后，第一，中间）：江恩项目摘要临界肢体缺血（CLI）的糖尿病患者的肢体截肢和死亡率显着高。 CLI以极低的血液灌注和退化的骨骼肌为特征。因此，再生脉管系统和骨骼肌将挽救四肢。然而，内皮和骨骼肌细胞的存活不良糖尿病CLI的高血糖和缺血状态下的下细胞功能会损害肢体修复。当前，尽管生长因子疗法代表了一种有希望的策略，但尚无有效的治疗方法。但是，生长因子治疗在再生脉管系统和骨骼的再生方面具有相对较低的治疗疗效肌肉，作为血管形成和肌发生同时需要多种生长因素，这些因素不能很容易通过当前的方法传递。在这个项目中，我们建议使用具有促血管生成和促生物基因的新型TRIM72蛋白在糖尿病性CLI中再生脉管系统和骨骼肌。 TRIM72将设计为更长的缺血组织中的保留时间（扩散速率较慢），因此表现出更长的治疗作用。交付设计的TRIM72（ETRIM72），它将封装在缺血性肢体靶向纳米颗粒中，其次是通过临床上有吸引力的静脉注射递送。然后，纳米颗粒将主要积聚在缺血性四肢并逐渐释放Etrim72。释放的蛋白质将通过（1）促进血管化和肌发生通过细胞膜修复改善内皮细胞和骨骼肌细胞的存活，并激活细胞生存激酶；（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，我们通过遗传融合的TRIM72与肽CSMLKAC开发了ETRIM72 该靶向缺血性四肢的缺血环境。 Etrim72的第一个版本能够保留在缺血性中四肢明显长于TRIM72。注射静脉注射缺血性肢体靶向后，ETRIM72释放纳米颗粒，释放的ETRIM72显着促进了糖尿病患者血管和骨骼肌的再生缺血性四肢。 TRIM72在高血糖和促进血管形成和肌发生中的功能缺血状况以前没有报道。基于我们的初步研究，我们假设受控的ETRIM72的释放将同时增加内皮和骨骼肌细胞存活，迁移和形态发生在高血糖和缺血下糖尿病缺血性肢体中脉管系统和骨骼肌的加速加速。 AIM＃1将检验以下假设：最佳ETRIM72释放曲线将显着促进生存，迁移在高葡萄糖和缺血性条件下，内皮细胞和成肌细胞的形态发生。 AIM＃2将测试使用糖尿病鼠肢体缺血模型，ETRIM72释放纳米颗粒的功效。该项目具有创新性，因为它正在设计一种新型的促血管生成和临时蛋白糖尿病性缺血性肢体中同时再生脉管系统和骨骼肌。较长的组织保留率工程蛋白的时间以及局部和受控释放的时间有望显着改善治疗功效。 OMB No. 0925-0001/0002（Rev. 03/2020通过02/28/2023批准）页面延续格式页面