Quantitative analysis of the evolving genotype-to-phenotype map

不断演变的基因型到表型图谱的定量分析

基本信息

批准号：
8600700
负责人：
Daniel Jarosz
金额：
$ 24.27万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8600700
关键词：
Address Adopted Affect Assimilations Bacteria Biochemistry Carbon Cell Cycle Cells Chemicals Clinical Collaborations Communication Complex DNA Damage DNA damage checkpoint Dependence Disease Disease remission Distant Drug resistance Enabling Factors Environment Evolution Fungal Drug Resistance Genes Genetic Genetic Polymorphism Genetic Techniques Genetic Variation Genotoxic Stress Genotype Growth Homeostasis Human Pathology Infection Institutes Maintenance Malignant Neoplasms Maps Mentors Messenger RNA Molecular Molecular Chaperones Molecular Conformation Multi-Drug Resistance Mutate Mutation NADH dehydrogenase (ubiquinone)Nerve Degeneration Oncogenes Oncogenic Oxidative Stress Parents Pharmaceutical Preparations Phenotype Phosphotransferases Prions Process Proliferating Protein Conformation Protein Structure Initiative Proteins Proteome Quantitative Genetics Regulatory Element Research Resistance Ribosomes Saccharomyces cerevisiae Sentinel Shapes Signal Pathway Signal Transduction Source Stress System Techniques Therapeutic Intervention Toxic effect Training Variant Yeasts antimicrobial bacterial genetics base cancer addiction cancer cell cancer therapy career cohort digital environmental change genetic regulatory protein genome wide association study insight next generation sequencing novel protein folding protein function reconstruction resistance mechanism response trait transcription factor yeast genetics

项目摘要

PROJECT SUMMARY Oncogene-directed cancer treatments and antimicrobial therapies are often thwarted by rapid acquisition of drug resistance, a chief barrier to lasting remission. Promising insight is emerging from a seemingly distant field - protein folding. To function, proteins must adopt complex, often metastable conformations. Perilously, many diseases arise from folding or misfolding of a single protein. My previous studies have focused on Hsp90, a molecular chaperone that folds metastable proteins critical for oncogenic transformation and signaling. By influencing the fold and function of an elite cohort of regulatory proteins, this chaperone has the power to influence the evolution of new traits. I will address the following aims during my remaining mentored training and initial independent research career: (I) Determine how Hsp90 transforms genetic variation. Hsp90 strongly impacts the effects of polymorphisms in Mec1/ATR, a central player in cancer signaling, enabling responses to certain genotoxic stresses at the expense of others. Biochemically and functionally, I will examine how this affects the DNA damage response, cell cycle, and viability. Additionally, is will investigate how Hsp90 impacts the transcriptional network of Pdr8, a transcription factor that enables resistance to many drugs. Finally, I will examine how Hsp90 inhibition transforms the effects of polymorphisms in cis-regulatory elements of NDI1, to create strong resistance to oxidative stresses. (II) Investigate assimilation of Hsp90-contingent phenotypes. To investigate eventual breakthrough drug resistance I will isolate causative variation initially and after assimilation (when resistance is Hsp90-independent). High-throughput genetic techniques and next-generation sequencing will provide a mechanistic understanding of this phenomenon. (III) Identify and characterize additional factors that allow highly mutated cells to survive, proliferate, and evolve new traits. Cancer cells must sustain massive mutation loads and consequently toxic proteome destabilization. I will screen for proteins that rescue growth of highly mutated strains but do not affect unmutated parents. (IV) Identify and characterize additional protein-based mechanisms that facilitate adaptation to new environments. Using digital ribosome profiling, yeast and bacterial genetics, and biochemistry I will characterize two prions, [PSI+] and [GAR+], and determine how they affect adaptation to changing environments. The results of these studies will offer detailed insight into how Hsp90, and protein homeostasis more generally, controls signaling pathways that enable drug resistance and how these mechanisms contribute to breakthrough resistance. They will also expose an Achilles' heel common to all cancers - addiction to factors that enable maintenance of massive mutation loads.

项目概要癌基因导向的癌症治疗和抗菌疗法常常因快速的治疗而受阻获得耐药性是持久缓解的主要障碍。有希望的见解正在出现来自一个看似遥远的领域——蛋白质折叠。为了发挥作用，蛋白质必须采用复杂的、通常是亚稳态构象。危险的是，许多疾病都是由单个基因的折叠或错误折叠引起的。蛋白质。我之前的研究主要集中在 Hsp90，一种折叠的分子伴侣亚稳态蛋白对于致癌转化和信号转导至关重要。通过影响折叠和一群精英调节蛋白的功能，该伴侣有能力影响新特征的演变。在我剩余的指导期间，我将实现以下目标培训和最初的独立研究生涯：（一）确定Hsp90如何转化遗传变化。 Hsp90 强烈影响 Mec1/ATR 多态性的影响，Mec1/ATR 是一个核心参与者在癌症信号传导中，能够以牺牲其他应激为代价来响应某些基因毒性应激。从生化和功能角度，我将研究这如何影响 DNA 损伤反应、细胞周期和活力。此外，我们还将研究 Hsp90 如何影响转录 Pdr8 网络是一种能够对多种药物产生耐药性的转录因子。最后，我会研究 Hsp90 抑制如何改变顺式调控中多态性的影响 NDI1 元素，对氧化应激产生强大的抵抗力。（二）调查 Hsp90 偶然表型的同化。研究最终突破性药物阻力我将在最初和同化之后隔离致病变异（当阻力是 Hsp90 独立）。高通量遗传技术和下一代测序将提供对这种现象的机械理解。（三）识别和表征允许高度突变的细胞生存、增殖和进化新细胞的其他因素特征。癌细胞必须承受大量的突变负荷，从而产生有毒的蛋白质组不稳定。我将筛选能够挽救高度突变菌株生长但不能恢复生长的蛋白质影响未突变的父母。 (IV) 识别和表征其他基于蛋白质的促进适应新环境的机制。使用数字核糖体分析、酵母和细菌遗传学以及生物化学我将表征两种朊病毒，[PSI+] 和 [GAR+]，并确定它们如何影响对不断变化的环境的适应。结果这些研究将提供关于 Hsp90 和蛋白质稳态如何更多的详细见解一般来说，控制导致耐药性的信号通路以及这些机制如何有助于突破阻力。他们还将暴露所有人共有的致命弱点癌症——对能够维持大量突变负荷的因素的成瘾。