Plasmalemma permeability and necroptosis: New targets for intracerebral hemo

质膜通透性和坏死性凋亡：脑内血液的新靶标

基本信息

批准号：
8617306
负责人：
MICHAEL J WHALEN
金额：
$ 36.53万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8617306
关键词：
Acute Acute Brain Injuries Autologous Biochemical Biological Markers Blood Blood - brain barrier anatomy Brain Brain Edema CD95 Antigens Cell Death Cell Survival Cell membrane Cells Cerebral hemisphere hemorrhage Cessation of life Combined Modality Therapy Corpus striatum structure Data Development Dose Edema Employee Strikes FDA approved Fatal Outcome Formularies Functional disorder Genetic Hematoxylin and Eosin Staining Method Human In Situ Nick-End Labeling Injury Ischemic Stroke Knock-out Knockout Mice Label Lasers Mediating Mediator of activation protein Membrane Microscopy Modeling Motor Mus Necrosis Neurologic Dysfunctions Neurons Outcome Outcome Measure Patients Permeability Phenotype Physiologic pulse Play Propidium Diiodide Protocols documentation Public Health RIPK1 gene RIPK3 gene Reporting Role Signal Transduction Speed Staining method Stains TNF gene Testing Time Tissues Traumatic Brain Injury Traumatic cerebral hemorrhage Tumor Necrosis Factor-alpha Work YOYO-1 base brain cell cell injury cell type clinically relevant cohort collagenase combinatorial controlled cortical impact copolymer fluoro jade functional outcomes human TNF protein improved in vivo indexing inhibitor/antagonist injured motor function improvement new therapeutic target novel pill programs research study restoration small molecule therapeutic target translational approach vinyl acetate

项目摘要

DESCRIPTION (provided by applicant): Non-traumatic intracerebral hemorrhage (ICH) is a major public health problem that lacks specific therapy. We present several lines of evidence suggesting that plasmalemma permeability and necrosis contribute to cell death after collagenase induced ICH in mice. Following ICH, plasmalemma permeability to propidium iodide (PI) is a key feature of cellular injury and death; plasmalemma permeability after ICH is reduced by 50% in mice deficient in TNF alpha and Fas receptor; necrotic-like cell death, assessed by EM, histological, and biochemical criteria occurs after ICH; and PI+ cells are reduced after ICH by RIPK3 knockout (a key molecule governing programmed necrosis). These preliminary findings strongly suggest that necroptosis (initiated by TNF/Fas and mediated by RIPK1 and RIPK3) contribute to cell death, and perhaps neurological dysfunction, after ICH. Necrostatins are small molecule inhibitors of RIPK1 that inhibit necroptosis in cells and protect against cell death and functional deficits in brain trauma and ischemic stroke models. Kollidon VA64 is a FDA approved formulary component in pill manufacture that reseals injured cells after CCI and ICH in vivo. VA64 administration reduces blood brain barrier damage, brain edema, cell death, and motor dysfunction after CCI, and reseals injured cells and reduces BBB damage after collagenase ICH in mice. Using mouse collagenase and autologous blood ICH models, we propose 3 specific aims to test the central hypotheses that (1) plasmalemma damage is a marker and mediator of cell death after ICH, and (2) that programmed necrosis contributes to cell death and functional outcome after ICH. Aim 1 will use pulse labeling experiments to follow the fate of injured cells in vivo and test the hypothesis that loss of plasmalemma integrity is a biomarker of fatal cellular injury after ICH; Aim 2 will test the hypothesis that VA64 restores plasmalemma integrity, reduces secondary injury and rescues injured brain cells from death after ICH; Aim 3 will test the hypothesis that necroptosis induces loss of plasmalemma integrity and cell death after ICH, using RIPK3 knockout mice and necrostatins, and a translational approach using necrostatins plus VA64. Completion of these Aims will establish necrosis as central to ICH, and may establish VA64 and necrostatins as novel therapies for ICH.

描述（由申请人提供）：非创伤性脑内出血（ICH）是缺乏特定疗法的主要公共卫生问题。我们提供了几种证据，表明胶原酶在小鼠中诱导的ICH后的血浆渗透性和坏死有助于细胞死亡。在ICH之后，对碘化丙啶（PI）的血浆渗透性是细胞损伤和死亡的关键特征。在缺乏TNFα和Fas受体的小鼠中，ICH后的血浆渗透性降低了50％。由EM，组织学和生化标准评估的坏死样细胞死亡发生在ICH之后。通过RIPK3基因敲除ICH后，PI+细胞（PI CROVENDENDENED坏死）减少了。这些初步发现强烈表明坏死性（由TNF/FAS引发并由RIPK1和RIPK3介导）在ICH之后导致细胞死亡，也许会导致神经功能障碍。坏死蛋白是RIPK1的小分子抑制剂，可抑制细胞中坏死性，并预防脑创伤和缺血性中风模型中细胞死亡和功能缺陷。 Kollidon VA64是FDA批准的药丸制造中的配方组件，可在CCI和ICH体内重新密封受伤的细胞。 VA64给药可减少CCI后血液脑屏障损伤，脑水肿，细胞死亡和运动功能障碍，并重新密封受伤的细胞并减少小鼠胶原酶ICH后BBB损伤。使用小鼠胶原酶和自体血液ICH模型，我们提出了3个特定的目的，以测试（1）（1）浆膜损伤是ICH后细胞死亡的标记和介体，（2）编程坏死会导致细胞死亡和ICH后的功能性结果。 AIM 1将使用脉冲标记实验在体内遵循受伤细胞的命运，并检验假设血浆完整性的丧失是ICH后致命细胞损伤的生物标志物。 AIM 2将检验以下假设：VA64恢复血浆完整性，减少次要损伤并救出ICH后死亡的脑细胞受伤。 AIM 3将检验以下假设：使用RIPK3基因敲除小鼠和坏死蛋白在ICH后诱导血浆完整性和细胞死亡的丧失，以及使用Necrostatins和VA64的转化方法。这些目标的完成将确立坏死的核心，并可能建立VA64和坏死蛋白作为ICH的新疗法。