Expanding high-impact mentorship and research in the Bracht Laboratory

扩大布拉赫特实验室的高影响力指导和研究

• 批准号: 10792325
• 负责人: John Russell Bracht
• 金额: $ 31.36万
• 依托单位: AMERICAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-07 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10792325
• 关键词: American; Apoptosis; Arginine; Astrocytes; Autoimmune Diseases; Award; Caenorhabditis elegans; Cellular Stress; Complex; Data; Environment; Exhibits; Funding; Gene Expression; Genes; Genome; Gold; High temperature of physical object; Human Pathology; Hypoxia; Investigation; Laboratories; Link; Malignant Neoplasms; Mentors; Mentorship; Metabolic; Methane; Mitochondria; Molecular; Mutate; Nematoda; Nerve Degeneration; Neurodegenerative Disorders; Organism; Parents; Pathway interactions; Physiology; Process; Proteins; Proton Pump; Publishing; Pump; RNA Interference; Regulation; Research; Site; South Africa; South African; Starvation; Stress; Students; Tissues; Toxin; Tumor stage; Universities; Whole Organism; cell injury; cytochrome c oxidase; feeding; human disease; insulin signaling; mitochondrial genome; neurotrophic factor; parent grant; repaired; resilience; response; single cell sequencing; single-cell RNA sequencing; stress resilience; stressor; transcriptome sequencing

Abstract of Supplement: The proposed supplemental funding will be used to both expand mentoring of a diverse student research group in the Bracht laboratory at American University, and expand the research proposed in the parent grant (1R15GM146207). The parent grant abstract is reproduced below; the supplemental research is within the scope of this parent grant. Abstract of parent award: A critical and well-studied cellular stress response pathway, the Unfolded Protein Response (UPR), protects organisms against several stressors including heat, hypoxia, starvation, and toxins. Helping to repair cellular damage, the UPR can also trigger apoptosis if the stress is ongoing, severe, and unrecoverable. Therefore, proper regulation of this pathway is essential, particularly since its malfunction contributes to human pathologies including autoimmune disorders, cancer, and neurodegenerative diseases. The Bracht lab recently published the genome of a nematode, Halicephalobus mephisto, isolated from the deep terrestrial subsurface of South Africa, over a kilometer underground. This organism has adapted to a stressful environment: hot, hypoxic, and rich in methane. Therefore the organism displays a naturally evolved resilience to stresses that would normally cause lethality; we also found that its UPR pathway is a site where adaptation has occurred. We have confirmed that RNA Interference (RNAi) by feeding can be used to modulate gene expression in this organism, setting the stage for a molecular investigation of stress resilience. New supplemental aims: Aim 1. Single-cell RNA-seq of ARMET RNAi in H. mephisto. In the parent grant, we described a gene, Arginine-Rich, Mutated in Early-stage Tumors / Mesencephalic Astrocyte derived Neurotrophic Factor (ARMET/MANF), that is highly expressed when the nematodes are grown at high temperatures. Surprisingly, our RNAseq data have uncovered an exciting and previously unknown link between the UPR pathway and insulin signaling. We propose to perform single-cell sequencing to identify which tissues are implicated in this process. Aim 2. Analysis of a more efficient mitochondrial proton pump in H. mephisto. In this aim we extend the research into H. mephisto adaptation into hypoxia, an environmental stress described in the parent grant. We found that the mitochondrial genome of H. mephisto exhibits an altered proton pump in the cytochrome c oxidase complex. We show that this pump is more effective than that encoded by C. elegans, a standard laboratory nematode, non-hypoxia adapted worm. We propose to perform extensive metabolic analysis of H. mephisto to confirm and extend these findings to whole- organism physiology.

补充摘要：拟议的补充资金将用于扩展 在美国大学的Bracht实验室中指导一个多元化的学生研究小组， 扩展父母赠款（15GM146207）中提出的研究。父母赠款摘要是 在下面复制；补充研究在该父母赠款的范围内。 父母奖励摘要：一个关键且研究良好的细胞应力反应途径， 展开的蛋白质反应（UPR）可保护生物免受多种压力源，包括热，缺氧， 饥饿和毒素。有助于修复细胞损伤，如果应力，UPR也会触发凋亡 正在进行，严重且无法恢复。因此，对这一途径的适当调节至关重要， 特别是因为它的故障导致了包括自身免疫性疾病在内的人类病理，所以 癌症和神经退行性疾病。 Bracht Lab最近发布了线虫的基因组， 与南非深层地下隔离的Halicephalobus Mephisto超过一公里 地下。这种生物已经适应了压力大的环境：热，低氧且富含甲烷。 因此，生物体表现出对通常会导致的压力的自然发展的弹性 致死我们还发现其UPR途径是发生适应性的站点。我们已经确认 通过喂养的RNA干扰（RNAi）可用于调节该生物体中的基因表达， 为压力弹性的分子研究奠定了基础。 新的补充目的： AIM1。H.Mephisto中的RNAi的单细胞RNA-seq。 在父授予中，我们描述了一个基因，富含精氨酸的基因，在早期肿瘤 / 中脑星形胶质细胞衍生的神经营养因子（Ambet/manf），高度表达 线虫在高温下生长。令人惊讶的是，我们的RNASEQ数据发现了 UPR途径和胰岛素信号传导之间令人兴奋且以前未知的联系。我们建议 执行单细胞测序，以识别哪些组织与此过程有关。 AIM 2。分析H. Mephisto中更有效的线粒体质子泵。 在此目的中，我们将研究扩展到梅菲斯托链球菌适应缺氧，这是一种环境压力 在父母赠款中描述。我们发现，梅菲斯托的线粒体基因组表现出改变 细胞色素C氧化酶复合物中的质子泵。我们表明，该泵比这更有效 由秀丽隐杆线虫（标准实验室线虫）编码，非催眠素适应蠕虫。我们建议 对H. Mephisto进行广泛的代谢分析，以确认并将这些发现扩展到整个 有机体生理。