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

• 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
• 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

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）：癌基因指导的癌症治疗和抗菌疗法通常会因迅速获取耐药性而挫败，这是持续缓解的主要障碍。有希望的见解正在从看似遥远的场 - 蛋白质折叠中出现。为了起作用，蛋白质必须采用复杂的，通常是亚稳定的构象。危险的是，许多疾病是由于单个蛋白质的折叠或错误折叠引起的。我以前的研究集中在HSP90上，HSP90是一种分子伴侣，它折叠了对致癌转化和信号传导至关重要的可稳定蛋白。通过影响调节蛋白精英队列的褶皱和功能，该伴侣有能力影响新特征的演变。我将在剩余的受过指导培训和最初的独立研究职业中解决以下目标：（i）确定HSP90如何改变遗传变异。 HSP90强烈影响多态性在MEC1/ATR（癌症信号传导中的核心参与者）中的影响，从而使对某些遗传毒性应激的反应以牺牲他人为代价。从生化和功能上讲，我将研究这如何影响DNA损伤响应，细胞周期和活力。此外，将研究HSP90如何影响PDR8的转录网络，PDR8的转录因子具有对许多药物的耐药性。最后，我将研究HSP90抑制如何改变NDI1顺式调节元件中多态性的影响，从而产生对氧化应激的强烈抗性。 （ii）研究HSP90凸型表型的同化。为了研究最终的突破性耐药性，我将最初和同化后分离出病因变异（当电阻不依赖于Hsp90时）。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）识别并表征了允许高度突变细胞生存，增殖和发展新特征的其他因素。癌细胞必须维持巨大的突变负荷，因此有毒蛋白质组不稳定。我将筛选蛋白质，这些蛋白质可以挽救高度突变的菌株的生长，但不会影响未分离的父母。 （iv）识别并表征促进适应新环境的其他基于蛋白质的机制。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[PSI+]和[GAR+]的两个pr象，并确定它们如何影响对不断变化的环境的适应。这些研究的结果将为HSP90和蛋白质稳态如何更广泛地提供详细的见解，控制着信号通路，这些途径能够耐药性以及这些机制如何促进突破性的抵抗力。他们还将暴露出所有癌症常见的阿喀琉斯的脚跟 - 对能够维持大规模突变负荷的因素上瘾。 公共卫生相关性：最初的癌症指导癌症疗法和多种抗菌治疗的最初承诺通常会因迅速获得耐药性而挫败。了解使复杂系统能够在选择，增殖和发展的新特征中生存的分子机制将提供急需的见解，以了解如何克服这种持续缓解和消除疾病的障碍。