The role of proBDNF-p75NTR signaling in hind limb ischemia.

proBDNF-p75NTR 信号在后肢缺血中的作用。

基本信息

批准号：
10603645
负责人：
Katherine Aby
金额：
$ 3.98万
依托单位：
UNIVERSITY OF SOUTH DAKOTA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10603645
关键词：
Amputation Apoptosis Attenuated Binding Proteins Biomedical Research Blood flow Brain Brain-Derived Neurotrophic Factor C57BL/6 Mouse Cell Adhesion Molecules Central Nervous System Communication Contralateral Cyclic AMP-Dependent Protein Kinases Data Data Analyses Disease Doctor of Philosophy Endothelial Cells Endothelium Equipment Event Experimental Designs Extracellular Domain Functional disorder Growth Health Impairment In Vitro Inflammation Injury Ischemia Knock-out Knowledge Learning Ligation Limb structure Mediating Mentors Metabolism Modeling Mus Muscle Muscle function NGFR Protein Neurotrophic Tyrosine Kinase Receptor Type 2 Nitric Oxide Operative Surgical Procedures Pathologic Patients Peptide Hydrolases Perfusion Peripheral arterial disease Phosphorylation Play Prevention Production Productivity Proprotein Convertase 1 Protein Secretion Proteins Reperfusion Injury Reperfusion Therapy Research Role Signal Pathway Signal Transduction Skeletal Muscle Testing Tissues Tourniquets Trauma Vascular Diseases Vascular Endothelial Cell Vascular System Wild Type Mouse Work blood perfusion brain-derived neurotrophic factor precursor career cell injury disability endothelial dysfunction experience experimental study extracellular improved in vivo injured limb ischemia neuroinflammation neuron loss neuroprotection new therapeutic target novel overexpression preservation prevent receptor small molecule therapeutic target trans-Golgi Network vascular inflammation vascular injury

项目摘要

The role of proBDNF-p75NTR signaling in hind limb ischemia Brain-derived neurotrophic factor (BDNF) and its precursor proBDNF have been well characterized in their contrasting roles in neuroprotection and neuroinflammation in the central nervous system respectively. Recently, BDNF and proBDNF have been shown to exist in tissues outside of the central nervous system, but their role in these tissues is still unclear. In my study it was shown that whole muscle lysate contains high levels of proBDNF and relatively low levels of mature BDNF compared to whole brain lysate. This project aims to investigate the functional role of skeletal muscle-derived proBDNF in hind limb ischemia-reperfusion (IR) injury. In this model, in which C57BL/6 mice are subjected to ligation of the right hind limb, proBDNF and its receptor p75NTR were both upregulated, and p75NTR downstream signaling pathways were activated in the ischemic muscle compared to the contralateral control limb muscles. The data further showed that treatment with LM11A-31, a p75NTR moderator, following IR injury improved perfusion of the ischemic limb compared to untreated IR controls, indicating improved endothelial function. Moreover, skeletal muscle-specific knockout of BDNF significantly improved the blood perfusion in the ischemic limb compared to wild-type mice. Previous studies have suggested that endothelial p75NTR may mediate vascular injury. Based on these results, we hypothesize that the upregulated proBDNF in skeletal muscle following IR injury may stimulate endothelial p75NTR, promoting cleavage of the intercellular domain (ICD) and subsequent inhibition of PKA-induced phosphorylation of eNOS and nitric oxide production leading to vascular dysfunction and inflammation. To test this hypothesis, mice with skeletal muscle-specific knockout of BDNF will be used to determine the role of muscle-derived proBDNF in IR injury. Then mice with endothelial-specific knockout of p75NTR will be used to examine whether endothelial p75NTR mediates IR injury. Finally, in vitro and in vivo experiments will be used to define the inhibition of PKA by the ICD of p75NTR and subsequent eNOS function being responsible for the IR injury. The results of this project will reveal a novel mechanism underlying IR injury and suggest new targets for the prevention and treatment of IR injury in patients. While under the superior intellectual guidance and support of a mentor with over 20 years of experience in the field, functional knowledge of experimental design, data analysis and interpretation, and scientific communication will be developed, leading to Ph.D. and enabling a successful and productive career in biomedical research. Experiments will be carried out using top-of-the-line research equipment in premier facilities with ample opportunities for learning, growth, and support.

proBDNF-p75NTR信号在后肢缺血中的作用脑源性神经营养因子 (BDNF) 及其前体 proBDNF 已在它们分别在中枢神经系统的神经保护和神经炎症中发挥对比作用。最近，BDNF 和 proBDNF 已被证明存在于中枢神经系统以外的组织中，但它们在这些组织中的作用仍不清楚。在我的研究中表明，整个肌肉裂解物含有高与全脑裂解物相比，proBDNF 水平和成熟 BDNF 水平相对较低。该项目旨在研究骨骼肌来源的 proBDNF 在后肢缺血再灌注 (IR) 中的功能作用受伤。在该模型中，C57BL/6 小鼠的右后肢被结扎，proBDNF 及其受体p75NTR均上调，并且p75NTR下游信号通路在缺血肌肉与对侧对照肢体肌肉相比。数据进一步表明，治疗与 LM11A-31（一种 p75NTR 调节剂）相比，IR 损伤后改善了缺血肢体的灌注未经处理的 IR 对照，表明内皮功能得到改善。此外，骨骼肌特异性敲除与野生型小鼠相比，BDNF 显着改善了缺血肢体的血液灌注。以前的研究表明内皮p75NTR可能介导血管损伤。根据这些结果，我们假设 IR 损伤后骨骼肌中 proBDNF 的上调可能会刺激内皮细胞 p75NTR，促进细胞间结构域 (ICD) 的裂解并随后抑制 PKA 诱导的 eNOS 磷酸化和一氧化氮生成导致血管功能障碍和炎症。测试根据这一假设，骨骼肌特异性敲除 BDNF 的小鼠将被用来确定 IR 损伤中肌肉衍生的 proBDNF。然后，内皮特异性敲除 p75NTR 的小鼠将被用于检查内皮 p75NTR 是否介导 IR 损伤。最后，将使用体外和体内实验定义 p75NTR 的 ICD 和随后的 eNOS 功能对 PKA 的抑制作用红外线损伤。该项目的结果将揭示 IR 损伤的新机制并提出新的靶点用于预防和治疗患者IR损伤。在上级智力指导和拥有超过 20 年该领域经验、实验设计功能知识的导师的支持，将发展数据分析和解释以及科学交流，从而获得博士学位。并启用在生物医学研究领域取得成功且富有成效的职业生涯。实验将使用顶级的一流设施中的研究设备，提供充足的学习、成长和支持的机会。