Role of PINCH in neuronal response to HIV infection of the CNS

PINCH 在中枢神经系统 HIV 感染的神经元反应中的作用

基本信息

批准号：
7685556
负责人：
Dianne Teresa LANGFORD
金额：
$ 35.63万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7685556
关键词：
Address Amino Acid Sequence Binding Biological Brain Brain Injuries Cells Central Nervous System Diseases Communication Cysteine Data Dementia Disease Disease Progression Exposure to Extracellular Matrix Focal Adhesions Goals Growth Factor HIV HIV Infections Hand Impaired cognition Impairment In Vitro Individual Infection Inflammatory Injury Integration Host Factors Integrins Life Mediating Neurites Neuroglia Neurologic Neurons Nuclear Nuclear Export Pathway interactions Patients Pattern Peptide Sequence Determination Peptide Signal Sequences Pharmaceutical Preparations Play Post-Translational Protein Processing Protein Isoforms Proteins Quality of life Recovery Recruitment Activity Reporting Research Role Severities Signal Pathway Signal Transduction Site Site-Directed Mutagenesis Small Interfering RNA Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Synapses Testing Time Urticaria Viral Viral Proteins adapter protein brain cell fitness histidine-rich proteins improved integrin-linked kinase interest laser capture microdissection neuron loss neuronal survival protein complex protein expression protein function public health relevance release factor repaired response trafficking

项目摘要

DESCRIPTION (provided by applicant): The long-term objective is to characterize mechanisms by which a host-derived protein called PINCH promotes neuronal fitness in response HIV infection of the CNS with the overall goal to understand PINCH's role in neurons exposed to viral and inflammatory factors. Synaptodendrtic injury correlates closely with the presence and severity of cognitive impairment in HIV patients. Since synaptodendritic damage in HIV infection of the brain is partially reversible and can occur in the absence of significant neuronal loss, information gained from these studies will enhance our ability to improve therapies to treat HIV infection of the brain and alleviate neurological complications observed in many HIV patients. In the brains of HIV infected individuals, PINCH is expressed exclusively by neurons showing signs of synaptodendritic damage. PINCH is an adapter protein that mediates bidirectional signal transduction between the extracellular matrix and intracellular networking pathways via interactions with integrin linked kinase (ILK) and Nck2. Thus, our overall hypothesis is that in response to disruptions in neuronal signaling caused by HIV infection of the CNS, PINCH protein is stabilized by sumoylation to promote proper interactions with ILK and Nck2. We propose two AIMS that investigate in vitro I) mechanisms by which PINCH expression is regulated, and II) the biological consequences of PINCH expression on neurons exposed to viral protein and host inflammatory factors produced during HIV infection of the CNS. Primary neurons treated with TNF-a or Tat to mimic some aspects of HIV infection of the brain will be assessed for PINCH expression via qRT-PCR, and Northern and Western analyses, and by siRNA knockdown. Sumoylation of PINCH will be assessed by reciprocal IP and Western analyses after exposure to TNFa and/or Tat. We further hypothesize that disruptions in PINCH's normal communication with ILK and Nck2 may diminish neuronal recovery. Neurons exposed to TNFa and/or Tat will be assessed for ILK and Nck2 expression and functional interactions with PINCH. To address biological consequences of PINCH in the context of HIV infection of the CNS, site-directed mutagenesis, PINCH protein accumulation and MALDI-TOF analyses will be used. Expanded studies of HIV patients' brains with MCMD, HIVE, dementia and importantly HAND in the absence of HIVE as well as HIV patients who died without CNS complications will be analyzed via laser capture microdissection of neurons and MALDI-TOF analyses will also be conducted. PUBLIC HEALTH RELEVANCE: RELEVANCE HIV patients are much living longer due to effective anti-HIV medications. Anti-HIV medications are less effective at treating HIV in the brain. So, many HIV patients suffer from neurological impairments. In this context, PINCH protein may contribute to neuron recovery during HIV infection of the brain. Understanding mechanisms by which PINCH functions will contribute to improved therapies to protect the brain from damage by HIV, and may improve the quality of life in HIV patients.

描述（由申请人提供）：长期目标是描述一种称为 PINCH 的宿主衍生蛋白促进神经元适应中枢神经系统 HIV 感染的机制，总体目标是了解 PINCH 在暴露于病毒和炎症的神经元中的作用因素。突触树突损伤与 HIV 患者认知障碍的存在和严重程度密切相关。由于大脑 HIV 感染中的突触树突损伤是部分可逆的，并且可以在没有明显神经元损失的情况下发生，因此从这些研究中获得的信息将增强我们改进治疗大脑 HIV 感染的疗法并减轻在许多 HIV 中观察到的神经系统并发症的能力患者。在 HIV 感染者的大脑中，PINCH 只由表现出突触树突损伤迹象的神经元表达。 PINCH 是一种接头蛋白，通过与整合素连接激酶 (ILK) 和 Nck2 的相互作用介导细胞外基质和细胞内网络通路之间的双向信号转导。因此，我们的总体假设是，为了应对中枢神经系统 HIV 感染引起的神经元信号传导中断，PINCH 蛋白通过苏酰化作用得以稳定，以促进与 ILK 和 Nck2 的正确相互作用。我们提出了两个体外研究的目标：I) PINCH 表达的调节机制，以及 II) PINCH 表达对暴露于 HIV 感染中枢神经系统期间产生的病毒蛋白和宿主炎症因子的神经元的生物学影响。用 TNF-a 或 Tat 处理的原代神经元模拟大脑 HIV 感染的某些方面，将通过 qRT-PCR、Northern 和 Western 分析以及 siRNA 敲低来评估 PINCH 表达。暴露于 TNFa 和/或 Tat 后，将通过相互 IP 和 Western 分析来评估 PINCH 的苏酰化。我们进一步假设 PINCH 与 ILK 和 Nck2 的正常通讯中断可能会削弱神经元的恢复。将评估暴露于 TNFa 和/或 Tat 的神经元的 ILK 和 Nck2 表达以及与 PINCH 的功能相互作用。为了解决中枢神经系统 HIV 感染背景下 PINCH 的生物学后果，将使用定点突变、PINCH 蛋白积累和 MALDI-TOF 分析。对患有 MCMD、HIVE、痴呆症和重要的 HAND 的 HIV 患者大脑以及没有 CNS 并发症而死亡的 HIV 患者的大脑的扩展研究将通过神经元的激光捕获显微切割进行分析，并且还将进行 MALDI-TOF 分析。公共卫生相关性：相关性由于有效的抗艾滋病毒药物，艾滋病毒患者的寿命更长。抗艾滋病毒药物在治疗大脑中的艾滋病毒方面效果较差。因此，许多艾滋病毒患者患有神经系统损伤。在这种情况下，PINCH 蛋白可能有助于 HIV 感染大脑期间的神经元恢复。了解 PINCH 功能的机制将有助于改进治疗方法，保护大脑免受 HIV 损害，并可能改善 HIV 患者的生活质量。