Expanding high-impact mentorship and research in the Bracht Laboratory

补充摘要：拟议的补充资金将用于扩大 指导美国大学Bracht实验室的一个多元化学生研究小组， 扩大在父母补助金（1 R15 GM 146207）中提出的研究。家长补助金摘要是 转载如下;补充研究是在这个母基金的范围内。 家长奖摘要：一个关键的和充分研究的细胞应激反应途径， 未折叠蛋白质反应（UPR），保护生物体免受几种应激因素的影响，包括热，缺氧， 饥饿和毒素帮助修复细胞损伤，UPR也可以触发细胞凋亡，如果压力 正在持续严重且无法恢复因此，对该途径的适当调节是至关重要的， 特别是因为它的功能障碍会导致包括自身免疫性疾病在内的人类疾病， 癌症和神经退行性疾病。Bracht实验室最近发表了一种线虫的基因组， Halicebusmephisto，从南非的深层地表下分离出来，超过一公里 地下这种生物已经适应了一个紧张的环境：炎热，缺氧，富含甲烷。 因此，生物体表现出自然进化的弹性，以应对通常会导致 致命性;我们还发现它的UPR途径是一个发生适应的位点。我们已经证实 通过进食的RNA干扰（RNAi）可以用来调节这种生物体中的基因表达， 为压力恢复的分子研究奠定了基础。 新的补充目标： 目标1. ARMET RNAi在H.墨菲斯托 在母基金中，我们描述了一种基因，富含精氨酸，在早期肿瘤中突变。 中脑星形胶质细胞源性神经营养因子（ARMET/MANF），当 线虫在高温下生长。令人惊讶的是，我们的RNAseq数据已经发现了一个 令人兴奋的和以前未知的联系之间的UPR途径和胰岛素信号。我们建议 进行单细胞测序，以确定哪些组织涉及这一过程。 目标2. H中更有效的线粒体质子泵的分析。墨菲斯托 为此，我们将研究扩展到H. mephisto适应缺氧，一种环境压力 在家长补助金中描述。我们发现H.梅菲斯特表现出一种改变了的 细胞色素c氧化酶复合物中的质子泵。我们证明了这种泵比那种泵更有效 由C. elegans，一种标准的实验室线虫，非缺氧适应蠕虫。我们建议 对H进行广泛的代谢分析Mephisto确认并将这些发现扩展到整个- 生物生理学

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.