Role of CNPase in TBI

CNPase 在 TBI 中的作用

基本信息

批准号：
8102066
负责人：
EDWIN Kerry JACKSON
金额：
$ 19.2万
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：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8102066
关键词：
2&apos ,3&apos -Cyclic-Nucleotide Phosphodiesterases 2&apos -adenylic acid Adenine Adenine Nucleotides Adenosine Autacoids Behavioral Paradigm Binding Biochemical Biological Assay Brain Brain Injuries Cell membrane Cells Cerebrospinal Fluid Cleaved cell Cognitive Collaborations Contusions Corpus Callosum Cyclic AMP Data Dorsal Electrons Enzymes Functional disorder Goals Grant Harvest High Pressure Liquid Chromatography Hippocampus (Brain)Histologic Human Impairment In Vitro Injury Inosine Investigation Isomerism Journals Kidney Knockout Mice Left Length Life Liquid substance Measures Mediating Memory impairment Messenger RNA Microdialysis Military Personnel Molecular Weight Morbidity - disease rate Motor Mus Myelin Neuroprotective Agents Neurotoxins Nucleotidases Oligodendroglia Outcome Patients Pharmacology Physiology Play Population Positioning Attribute Process Property Proteins Publications Purines RNA Research Personnel Research Project Grants Ribonucleases Role Sampling Second Messenger Systems Sensitivity and Specificity Source Structure TBI Patients Tail Testing Tissues Traumatic Brain Injury Work base cognitive function controlled cortical impact drug mechanism extracellular functional outcomes gray matter in vivo innovation innovative technologies interest interstitial meetings mitochondrial permeability transition pore mortality motor impairment neuroprotection new technology novel strategies nucleotidase phosphodiester phosphoric diester hydrolase prevent public health relevance purine quantum receptor research study second messenger tandem mass spectrometry white matter

项目摘要

DESCRIPTION (provided by applicant): The term "cAMP" is used universally to refer to 3',5'-cyclic adenosine monophosphate (3',5'- cAMP), the famous "second messenger" discovered by Dr. Earl Southerland. Importantly, the vast majority of investigators measure 3',5'-cAMP using various commercially available assay kits, and our group is the ONLY group in the world that routinely measures 3',5'-cAMP using high performance liquid chromatography-tandem mass spectrometry(LC-MS/MS). Dr. Jackson's lab focuses mostly on the role of purines in renal physiology/pharmacology, and routinely employs LC-MS/MS, rather than assay kits, to measure 3',5'-cAMP. In experiments unrelated to the brain, Dr. Jackson's lab serendipitously discovered that the kidney produces more 2',3'-cAMP (a positional isomer of 3',5'- cAMP) than 3',5'-cAMP. Dr. Kochanek's lab focuses primarily on mechanisms of traumatic brain injury (TBI), and has discovered that purines play a major role in protecting the brain from TBI. Because of Dr. Jackson's expertise in measuring purines and Dr. Kochanek's interest in the role of purines in TBI, a natural and productive collaboration evolved between these investigators. After the serendipitous discovery of 2',3'-cAMP in kidneys, Drs. Jackson and Kochanek decided to investigate whether 2',3'-cAMP exists in the brain. In preliminary studies, LC-MS/MS analysis of 44 samples of cerebral spinal fluid (CSF) from TBI patients showed that 2',3'-cAMP is a major constituent of "cAMP" in CSF from patients with TBI. In addition to 2',3'-cAMP, the levels of 2'-AMP, adenosine and inosine (metabolite of adenosine) were also measured in these same samples. Importantly, there was an large and significant correlation between 2',3'-cAMP and 2'-AMP, 2'-AMP and adenosine and 2'-AMP and inosine in human CSF. These surprising discoveries suggested a critical need to explore the role of 2',3'-cAMP in TBI pathophysiology, particularly with respect to the role of 2',3'-cAMP as a precursor for adenosine. Accordingly, the purpose of this exploratory project is to begin to characterize the significance of 2',3'-cAMP in TBI by testing the innovative concept that 2',3'-cAMP is involved in a "CNPase Neuroprotection Mechanism" in which brain injury leads to release of 2',3'-cAMP from mRNA, and 2',3'-cAMP is then metabolized to 2'-AMP by CNPase followed by conversion of 2'-AMP to adenosine. PUBLIC HEALTH RELEVANCE: TBI is a major source of mortality, morbidity and life-long impairment in both the civilian and military populations, with limited treatment options and generally dismal outcomes. The CNPase Neuroprotective Mechanism could be extremely important in producing extracellular adenosine during TBI, thus providing this protective, "retaliatory" metabolite to mitigate cellular damage. Manipulation of this mechanism by drugs could provide novel approaches to prevent and treat TBI.

描述（由申请人提供）：术语“cAMP”普遍用于指代3',5'-环单磷酸腺苷（3',5'-cAMP），它是Earl Southerland博士发现的著名的“第二信使”。重要的是，绝大多数研究人员使用各种市售检测试剂盒测量 3',5'-cAMP，而我们小组是世界上唯一使用高效液相色谱-串联质量常规测量 3',5'-cAMP 的小组光谱分析（LC-MS/MS）。 Jackson 博士的实验室主要关注嘌呤在肾脏生理学/药理学中的作用，并且通常使用 LC-MS/MS（而不是检测试剂盒）来测量 3',5'-cAMP。在与大脑无关的实验中，Jackson 博士的实验室偶然发现肾脏产生的 2',3'-cAMP（3',5'-cAMP 的位置异构体）比 3',5'-cAMP 更多。 Kochanek 博士的实验室主要关注创伤性脑损伤 (TBI) 的机制，并发现嘌呤在保护大脑免受 TBI 侵害方面发挥着重要作用。由于 Jackson 博士在测量嘌呤方面的专业知识以及 Kochanek 博士对嘌呤在 TBI 中的作用的兴趣，这些研究人员之间形成了自然而富有成效的合作。在肾脏中偶然发现 2',3'-cAMP 后，Drs. Jackson 和 Kochanek 决定调查大脑中是否存在 2',3'-cAMP。在初步研究中，对来自 TBI 患者的 44 份脑脊液 (CSF) 样本进行 LC-MS/MS 分析表明，2',3'-cAMP 是 TBI 患者 CSF 中“cAMP”的主要成分。除了 2',3'-cAMP 之外，还在这些相同的样品中测量了 2'-AMP、腺苷和肌苷（腺苷代谢物）的水平。重要的是，人脑脊液中 2',3'-cAMP 与 2'-AMP、2'-AMP 与腺苷、2'-AMP 与肌苷之间存在很大且显着的相关性。这些令人惊讶的发现表明迫切需要探索 2',3'-cAMP 在 TBI 病理生理学中的作用，特别是 2',3'-cAMP 作为腺苷前体的作用。因此，这个探索性项目的目的是通过测试 2',3'-cAMP 参与“CNPase 神经保护机制”这一创新概念来开始表征 2',3'-cAMP 在 TBI 中的重要性。损伤导致 2',3'-cAMP 从 mRNA 中释放，然后 2',3'-cAMP 被 CNPase 代谢为 2'-AMP，随后 2'-AMP 转化为腺苷。公共卫生相关性：TBI 是平民和军人死亡、发病和终身损害的主要根源，治疗选择有限，结果普遍令人沮丧。 CNPase 神经保护机制对于 TBI 期间产生细胞外腺苷极其重要，从而提供这种保护性的“报复性”代谢物来减轻细胞损伤。通过药物操纵这一机制可以提供预防和治疗 TBI 的新方法。