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：
8166021
负责人：
Daniel Jarosz
金额：
$ 9万
依托单位：
WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
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 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8166021
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 addiction antimicrobial bacterial genetics base cancer cell cancer therapy career cohort digital environmental change genetic regulatory protein genome wide association study insight next generation novel protein folding protein function reconstruction resistance mechanism response trait transcription factor yeast genetics

项目摘要

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Initial the initial promise of oncogene-directed cancer therapies and diverse antimicrobial treatments is often thwarted by rapid acquisition of drug resistance. Understanding molecular mechanisms that enable complex systems to survive selections, proliferate, and evolve new traits will provide much needed insights into how this hurdle to lasting remission and disease eradication can be overcome.

描述（由申请人提供）：癌基因导向的癌症治疗和抗菌疗法常常因快速获得耐药性而受阻，这是持久缓解的主要障碍。有希望的见解正在从一个看似遥远的领域——蛋白质折叠——中浮现出来。为了发挥作用，蛋白质必须采用复杂的、通常是亚稳态的构象。危险的是，许多疾病都是由单一蛋白质的折叠或错误折叠引起的。我之前的研究主要集中在 Hsp90，这是一种分子伴侣，可折叠对致癌转化和信号转导至关重要的亚稳态蛋白。通过影响精英调节蛋白群体的折叠和功能，该伴侣有能力影响新性状的进化。在我剩余的指导培训和最初的独立研究生涯中，我将实现以下目标：（I）确定 Hsp90 如何改变遗传变异。 Hsp90 强烈影响 Mec1/ATR 多态性的影响，Mec1/ATR 是癌症信号传导的核心参与者，能够以牺牲其他应激为代价来响应某些基因毒性应激。我将从生化和功能角度研究这如何影响 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 Pdr8 的转录网络，Pdr8 是一种能够对多种药物产生耐药性的转录因子。最后，我将研究 Hsp90 抑制如何改变 NDI1 顺式调控元件多态性的影响，从而产生对氧化应激的强大抵抗力。 (II) 研究 Hsp90 偶然表型的同化。为了研究最终突破性的耐药性，我将在最初和同化后（当耐药性与 Hsp90 无关时）分离出致病变异。高通量遗传技术和下一代测序将提供对这一现象的机械理解。 (III) 识别和表征允许高度突变的细胞生存、增殖和进化新性状的其他因素。癌细胞必须承受大量的突变负荷，从而导致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影响未突变亲本的蛋白质。 (IV) 识别并表征其他有助于适应新环境的基于蛋白质的机制。利用数字核糖体分析、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。这些研究的结果将提供关于 Hsp90 以及更广泛的蛋白质稳态如何控制导致耐药性的信号通路以及这些机制如何促成突破性耐药的详细见解。他们还将暴露所有癌症共有的致命弱点——对能够维持大量突变负荷的因素的成瘾。公共卫生相关性：癌基因导向的癌症治疗和多种抗菌治疗的最初承诺常常因快速获得耐药性而受挫。了解使复杂系统能够在选择中生存、增殖和进化新性状的分子机制，将为如何克服这一持久缓解和根除疾病的障碍提供急需的见解。