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PARP-1 is a Negative Modulator of the Heart Shock Response

PARP-1 是心脏休克反应的负调节剂

基本信息

批准号：
7457944
负责人：
RAJESH K. ANEJA
金额：
$ 12.29万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-03 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7457944
关键词：
Adenosine Triphosphate Animal Model Animals Binding Biology Cells Cessation of life Chairperson Childhood Condition Critical Care Critically ill children DNA DNA Binding Data Development Plans Electrophoretic Mobility Shift Assay Environment Enzymes Eukaryotic Cell Faculty Fibroblasts Functional disorder Gene Silencing Genes Genetic Genetic Transcription Goals Grant Heart Heat shock proteins Heat-Shock Proteins 70 Heat-Shock Response Immunoprecipitation In Vitro Indium Investigation K-Series Research Career Programs Knockout Mice Lead Learning Light Link Lipopolysaccharides Liver Lung Mediating Medicine Mentors Mentorship Messenger RNA Mitochondria Modeling Molecular Biology Mus Nicotinamide adenine dinucleotide Nuclear Outcome PARP inhibition Pathogenesis Pediatric Hospitals Play Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Production Promoter Regions Proteins RNA Research Personnel Research Project Grants Resistance Role Sepsis Septic Shock Series Shock Small Interfering RNA Stress Techniques Temperature Testing Training Transactivation Transcription Factor AP-2 Alpha Translational Research Universities Up-Regulation Wild Type Mouse biological adaptation to stress career cell type design heat shock transcription factor heat-shock factor 1 improved in vivo in vivo Model inhibitor/antagonist member novel poly(ADP-ribose)polymerase-1, mouse programs promoter protein expression repaired research study response transcription factor

项目摘要

DESCRIPTION (provided by applicant): I am a junior faculty member in the Department of Critical Care Medicine at the University of Pittsburgh. I am trained in Pediatric Critical Care Medicine and I recently completed a T-32 training grant at Cincinnati Children's Hospital. My short-term goal is to acquire additional training in molecular biology and learn additional techniques relevant to my long term goal: conduct translational research related to the biology of sepsis to improve outcomes for critically ill children. With the guidance of my mentor Dr. Mitchell P. Fink, (Chairman of my department at University of Pittsburgh) and key co-mentors, I have developed a program of formal coursework and tutorials that will form the educational and training core necessary to achieve my goals. My research project "Poly (ADP-ribose) polymerase (PARP) -1 is a negative modulator of the heat shock response" is designed to both provide vital new information and serve as an educational opportunity that compliments the other components of my career development plan. PARP-1 is a nuclear enzyme that upon activation mediates progression of septic shock. Genetic deletion or pharmacological inhibition of PARP is protective in animal models of sepsis. Another major cytoprotective stress response that is upregulated in critical care illness is the heat shock response (HSR). My preliminary data suggests that PARP-1 inhibition or genetic absence of PARP-1 is associated with upregulation of the HSR. DNA binding by the main transcription factor responsible for the HSR, heat shock factor (HSF)-1 in wild type (wt) cells is increased when heat-shocked cells are pretreated with a PARP-1 inhibitor (DIQ). Heat shocked PARP-/- cells demonstrate abundant HSP-70 expression and HSP-70 promoter activity, hallmarks of the HSR. However, DNA binding of HSF-1 in heat-shocked PARP-1-/- cells is not detectable by electrophoretic mobility shift assay (EMSA). Some of the experiments outlined in the present proposal should shed some light on this paradox (i.e. increased transcription of a HSF-1-dependent gene but absent HSF-1 DNA binding in heat shocked PARP-1-/- cells). Thus, in Aim 1,1 will confirm the data obtained in PARP-/- cells by studying wt cells transfected with small interfering RNA (siRNA) directed against PARP-1. Aim 2 examines the effect of PARP-1 inhibition on HSF-1-mediated transactivation. Aim 3 seeks to study these phenomena in an in vivo setting, specifically examining the effects of heat shock in wt and PARP-/- animals.

描述（由申请人提供）：我是匹兹堡大学重症监护医学系的一名初级教员。我接受过儿科重症监护医学培训，最近在辛辛那提儿童医院完成了 T-32 培训补助金。我的短期目标是获得分子生物学方面的额外培训，并学习与我的长期目标相关的其他技术：进行与脓毒症生物学相关的转化研究，以改善危重儿童的治疗结果。在我的导师米切尔·P·芬克博士（匹兹堡大学我系系主任）和主要共同导师的指导下，我制定了一个正式课程和教程计划，它将构成实现我的目标所需的教育和培训核心。我的研究项目“聚（ADP-核糖）聚合酶（PARP）-1 是热休克反应的负调节剂”旨在提供重要的新信息，并作为一个教育机会，补充我职业发展计划的其他组成部分。 PARP-1 是一种核酶，激活后可介导感染性休克的进展。 PARP 的基因缺失或药理学抑制对脓毒症动物模型具有保护作用。在重症监护疾病中上调的另一种主要细胞保护性应激反应是热休克反应（HSR）。我的初步数据表明 PARP-1 抑制或 PARP-1 基因缺失与 HSR 上调有关。当用 PARP-1 抑制剂 (DIQ) 预处理热休克细胞时，野生型 (wt) 细胞中负责 HSR 的主要转录因子热休克因子 (HSF)-1 与 DNA 的结合会增加。热休克的 PARP-/- 细胞表现出丰富的 HSP-70 表达和 HSP-70 启动子活性，这是 HSR 的标志。然而，电泳迁移率变动分析 (EMSA) 无法检测到热休克 PARP-1-/- 细胞中 HSF-1 的 DNA 结合。本提案中概述的一些实验应该可以阐明这一悖论（即热休克的 PARP-1-/- 细胞中 HSF-1 依赖性基因的转录增加但不存在 HSF-1 DNA 结合）。因此，在目标 1,1 中，将通过研究用针对 PARP-1 的小干扰 RNA (siRNA) 转染的 wt 细胞来确认在 PARP-/- 细胞中获得的数据。目标 2 检查 PARP-1 抑制对 HSF-1 介导的反式激活的影响。目标 3 旨在研究体内环境中的这些现象，特别是检查热休克对 wt 和 PARP-/- 动物的影响。