Resilience mechanisms of Arctic ground squirrel neurons

北极地松鼠神经元的恢复机制

• 批准号: 10550125
• 负责人: NEEL SINGHAL
• 金额: --
• 依托单位: VETERANS AFFAIRS MED CTR SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10550125
• 关键词: Acute; Acute Brain Injuries; Amino Acid Substitution; Amino Acids; Animal Model; Animals; Annexins; Apoptosis; Behavioral; Binding; Biochemical; Bioenergetics; Biological Assay; Biology; Blood flow; Brain Diseases; Brain Injuries; Brain Ischemia; CASP3 gene; COX7A2L Protein; COX7A2L gene; CRISPR/Cas technology; Candidate Disease Gene; Caring; Cell Death; Cell Survival; Cells; Cellular Assay; Cerebral Ischemia; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complex; Cytoprotection; Data; Development; Development Plans; Engineering; Evaluation; Exposure to; Female; Funding; Genes; Genetic; Genetic study; Glucose; Goals; Hand; Health; Hibernation; Human; Individual; Injury; Ischemia; Ischemic Stroke; K-Series Research Career Programs; Knock-in; Link; Mammals; Measurement; Measures; Mediating; Membrane Potentials; Mentors; Mentorship; Metabolic; Metabolic stress; Middle Cerebral Artery Occlusion; Mitochondria; Morbidity - disease rate; Mus; Nature; Neurons; Neuroprotective Agents; Outcome; Oxygen; Pathway interactions; Patient-Focused Outcomes; Permeability; Physicians; Physiological; Physiology; Positioning Attribute; Property; Proteins; Public Health; RIPK3 gene; Research; Research Personnel; Resistance; Role; Signal Pathway; Stroke; Techniques; Technology; Testing; Time; Toxin; Training; Training Programs; United States; Variant; Veterans; Visualization; Work; arctic ground squirrel; biological adaptation to stress; brain cell; cDNA Expression; career; career development; cellular resilience; clinical training; comparative genomics; complex IV; deprivation; disability; experience; experimental study; functional improvement; genome editing; improved; improved outcome; in vitro Model; in vivo; in vivo Model; insight; interest; male; metabolic phenotype; military veteran; mitochondrial membrane; mortality; nervous system disorder; neural; neuron loss; neuronal metabolism; neuronal survival; neuroprotection; neurovascular; new therapeutic target; novel; novel therapeutic intervention; oxidative damage; prevent; programs; promote resilience; protein expression; protein function; resilience; respiratory; statistics; stressor; stroke model; timeline; tool; translational potential; treatment strategy

Dr. Singhal's long-term goal is to be a VA physician investigator, elucidating mechanisms of neuronal bioenergetics in order to improve outcomes for veterans with acute brain injuries and other neurological diseases. The mechanisms linking cellular bioenergetics and cell death in ischemic brain diseases such as stroke are incompletely understood, and a greater understanding of these details will lead to the development of urgently needed neuroprotective agents for Veterans. During the Career Development Award-2 period, Dr. Singhal's goal is to acquire the training and implement the studies needed to understand and develop novel treatment strategies for conditions he treats in the Neurointensive care unit such as stroke. As such, he proposes a training program focused on identifying the mechanisms underlying the dramatic ischemia tolerance observed in one of nature's most resilient animals, the Arctic ground squirrel. Through comparative genomics and cell resilience-based cDNA expression screens, he discovered unique amino acid substitutions in cytoprotective proteins conferring increased resilience to metabolic stressors. The research proposed will build on his preliminary data and bring two candidate genes forward for rigorous study in neural cells using metabolic and cell death assays, visualization techniques, and biochemical measurements. He will use CRISPR-Cas9 knock-in technology to precisely edit the genes of interest, which are not fully characterized to date, and study the effect of their editing on neuronal cell resilience (Aim 1) and elucidate their cell physiologic mechanisms of action (Aim 2). Finally, he will test the candidate genes in the transient middle cerebral artery occlusion stroke model in vivo (Aim 3). The in-depth characterization using in vivo and in vitro models will establish a critical connection between the Arctic ground squirrel cytoprotective proteins and neuroprotective mechanisms, and importantly, provide valuable insights into novel drug targets. Dr. Singhal has assembled a diverse mentorship team comprised of experts in the fields of stroke, genetics, statistics, and mitochondrial physiology. Dr. Singhal's career development plan includes a clear timeline for individual tutorials with mentors and scientific advisors, hand-on experience, formal seminars, dissemination of findings, and plans for independent projects and funding. The mentored research and training in gene editing, neurovascular biology, mitochondrial physiology, and statistics described in this proposal will complement Dr. Singhal's training and facilitate his goal of launching an independent research career at the VA.

Singhal博士的长期目标是成为一名退伍军人事务部内科研究员，阐明神经元的机制 生物能量学，以改善患有急性脑损伤和其他神经疾病的退伍军人的预后。 在中风等缺血性脑疾病中，细胞生物能量学和细胞死亡之间的联系机制如下 不完全理解，而更多地了解这些细节将导致迫切的发展 退伍军人所需的神经保护剂。在职业发展奖-2期间，辛格尔博士的目标是 获得培训并实施所需的研究，以了解和开发新的治疗策略 他在神经重症监护病房治疗中风等疾病。因此，他提出了一项培训计划 重点研究了在自然界观察到的最严重的 北极地松鼠是一种有韧性的动物。通过比较基因组学和基于细胞弹性的cDNA 在表达筛查中，他发现细胞保护蛋白中独特的氨基酸替代增加了 对新陈代谢应激源的韧性。提议的研究将以他的初步数据为基础，并带来两名候选人 利用新陈代谢和细胞死亡分析、可视化技术和 生化测量。他将使用CRISPR-Cas9敲入技术来精确编辑感兴趣的基因， 到目前为止还没有完全表征的，并研究它们的编辑对神经细胞弹性的影响(目标1)和 阐明它们的细胞生理作用机制(目标2)。最后，他将测试候选基因在 短暂性大脑中动脉闭塞性卒中在体动物模型(目标3)。利用活体进行深入的表征 体外模型将在北极地松鼠细胞保护蛋白和 神经保护机制，重要的是，为新的药物靶点提供了有价值的见解。Singhal博士已经 组建了一个多元化的指导团队，由中风、遗传学、统计学和 线粒体生理学。Singhal博士的职业发展计划包括明确的个人教程时间表 导师和科学顾问，实践经验，正式研讨会，研究成果的传播，以及 独立的项目和资金。在基因编辑、神经血管生物学、 线粒体生理学和本提案中描述的统计数据将补充Singhal博士的培训和 促进他在退伍军人管理局开展独立研究事业的目标。