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 VA 医师研究员，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系统疾病的退伍军人的治疗结果。 缺血性脑部疾病（如中风）中细胞生物能学和细胞死亡之间的联系机制是 不完全理解，对这些细节的更好理解将导致迫切需要的开发 退伍军人需要神经保护剂。在职业发展奖 2 期间，Singhal 博士的目标是 获得理解和开发新的治疗策略所需的培训和实施研究 他在神经重症监护病房治疗中风等疾病。因此，他提出了一个培训计划 重点是确定在自然界中最严重的缺血耐受性之一中观察到的潜在机制 坚韧的动物，北极地松鼠。通过比较基因组学和基于细胞弹性的 cDNA 在表达筛选中，他发现细胞保护蛋白中独特的氨基酸取代可提高 对代谢压力源的抵抗力。拟议的研究将建立在他的初步数据的基础上，并带来两名候选人 使用代谢和细胞死亡测定、可视化技术和技术，将基因用于神经细胞的严格研究 生化测量。他将利用CRISPR-Cas9敲入技术来精确编辑感兴趣的基因， 迄今为止尚未完全表征，并研究它们的编辑对神经元细胞弹性的影响（目标 1）和 阐明其细胞生理作用机制（目标 2）。最后，他将测试候选基因 短暂性大脑中动脉闭塞体内卒中模型（目标 3）。使用体内的深入表征 体外模型将在北极地松鼠细胞保护蛋白和 神经保护机制，更重要的是，为新的药物靶标提供了有价值的见解。辛格尔博士有 组建了一支多元化的导师团队，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个人教程的明确时间表 导师和科学顾问、实践经验、正式研讨会、研究结果的传播和计划 独立的项目和资金。在基因编辑、神经血管生物学、 本提案中描述的线粒体生理学和统计数据将补充 Singhal 博士的培训和 促进他在退伍军人管理局开展独立研究生涯的目标。