Characterizing the Role of Leucyl Aminoacyl tRNA Synthetase (LARS) in Breast Cancer Metastasis

表征亮氨酰 tRNA 合成酶 (LARS) 在乳腺癌转移中的作用

• 批准号: 9909283
• 负责人: Maria Christina Passarelli
• 金额: $ 5.05万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-09 至 2023-12-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9909283
• 关键词: Academic Medical Centers; Acids; Affect; Amino Acids; Amino Acyl-tRNA Synthetases; Aminoacylation; Attenuated; Biogenesis; Biological Assay; Biology; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Treatment; Breast cancer metastasis; Cancer Biology; Cancer Etiology; Cancer Prognosis; Cell Line; Cell physiology; Cells; Charge; Clinical; Committee Members; Communication; Data; Development; Diagnosis; Disease; Doctor of Philosophy; Elements; Ensure; Environment; Enzymes; Foundations; Future; Genes; Genetic; Genetic Translation; Human; In Vitro; Internal Medicine; Knock-out; Knockout Mice; Knowledge; Leucine; Leucine-Specific tRNA; Literature; Malignant Neoplasms; Mediating; Medical; Mentors; Mentorship; Metastasis Suppression; Metastatic breast cancer; Modeling; Molecular; Molecular Target; Mouse Mammary Tumor Virus; Mus; Neoplasm Metastasis; Patient-Focused Outcomes; Patients; Phenotype; Physicians; Play; Process; Proteins; Proteomics; RNA; Research; Research Personnel; Residencies; Resources; Ribosomal RNA; Rice; Role; Scientist; Soft Agar Assay; Solid; Testing; Therapeutic; Training; Transfer RNA; Transfer RNA Aminoacylation; Translations; Tumor Biology; Tumor stage; Universities; Untranslated RNA; Woman; Work; Xenograft procedure; base; cancer cell; cancer subtypes; cell growth; combat; comparative; doctoral student; in vitro Assay; in vivo; insight; knock-down; malignant breast neoplasm; mortality; mouse model; neoplastic cell; novel; outcome forecast; overexpression; programs; ribosome profiling; skills; small hairpin RNA; therapeutic target; transcriptome sequencing; tumor; tumor initiation; tumor progression

PROJECT SUMMARY/ABSTRACT Breast cancer metastasis is a leading cause of mortality in US women. Despite advances in treatment for patients with early stage tumors, prognosis remains poor for patients with metastatic disease. Prior work has demonstrated a direct role for transfer RNAs (tRNAs) as drivers of breast cancer metastatic progression, through differential abundance resulting in changes in cell proteomic landscape. I hypothesize that aminoacylation status, as in, whether or not a tRNA is charged with an amino acid, and the requisite aminoacyl tRNA synthetase (aaRS) responsible for tRNA charging, will also play a role in breast cancer metastatic progression. To test this hypothesis, charged tRNAs were profiled across cells of differing metastatic potential, which identified key reductions in leucine tRNA charging in cells of higher metastatic potential. Preliminary xenograft and syngeneic mouse metastasis studies further identified leucyl aaRS (LARS), responsible for charging leucine tRNAs, as a metastasis suppressor. In this proposal, I will examine and further characterize the role of LARS in cancer metastasis. In Aim 1, I will further characterize the effects of LARS manipulation (1) through knockdown and overexpression in xenograft and syngeneic cancer mouse models, and (2) through genetic knockout in mouse models of breast cancer. In Aim 2, I will investigate (1) the cellular phenotype of metastasis suppression through in vitro assays and (2) the mechanism of charged tRNA-mediated metastasis suppression through downstream RNA sequencing analysis and ribosomal profiling. These proposed studies will fill a gap in literature: though tRNAs play a role in cancer progression, molecules related to their biogenesis have yet to be identified as therapeutically meaningful targets in cancer metastasis. Successful completion of these aims will lay the groundwork for future therapeutic targets with novel mechanisms, in treating metastatic breast cancer. I am an MD-PhD student at the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD program, completing the proposed aims in the lab of Dr. Sohail Tavazoie at Rockefeller University. Dr. Tavazoie, along with my committee members Drs. Charles Rice, Ping Chi and Kivanc Birsoy, provide strong scientific expertise, guidance and resources to ensure successful completion of the project. Together, we have taken steps to ensure my training plan also allows for development of necessary verbal and written communication skills as well as mentorship opportunities. My clinical mentors, Drs. Pamela Charney and Ping Chi, will offer critical advice on maintaining clinical acumen as I transition towards completion of medical training and select research-track residency programs in Internal Medicine. Successful completion of the proposed plan in this stellar training environment will enhance my foundational knowledge of RNA biology and teach me skills in cancer biology and mouse modeling, setting me solidly on the path towards becoming a physician scientist and independent investigator at an academic medical center.

项目概要/摘要 乳腺癌转移是美国女性死亡的主要原因。尽管治疗取得了进步 对于早期肿瘤患者，转移性疾病患者的预后仍然较差。之前的工作有 证明了转移 RNA (tRNA) 作为乳腺癌转移进展驱动因素的直接作用，通过 丰度差异导致细胞蛋白质组景观的变化。我假设氨酰化状态， 例如，tRNA 是否带有氨基酸，以及必需的氨酰 tRNA 合成酶 (aaRS) 负责 tRNA 充电，也将在乳腺癌转移进展中发挥作用。为了测试这个 假设，带电 tRNA 在不同转移潜能的细胞中进行了分析，从而确定了关键的 具有较高转移潜力的细胞中亮氨酸 tRNA 电荷减少。初步异种移植和同基因 小鼠转移研究进一步确定了负责亮氨酸 tRNA 充电的亮氨酰 aaRS (LARS) 转移抑制剂。在本提案中，我将研究并进一步描述 LARS 在癌症中的作用 转移。在目标 1 中，我将通过击倒和 在异种移植和同基因癌症小鼠模型中过度表达，以及（2）通过小鼠中的基因敲除 乳腺癌模型。在目标 2 中，我将研究 (1) 通过以下方式抑制转移的细胞表型： 体外测定和（2）带电 tRNA 介导的下游转移抑制机制 RNA 测序分析和核糖体分析。这些拟议的研究将填补文献空白：尽管 tRNA 在癌症进展中发挥作用，与其生物发生相关的分子尚未被确定为 癌症转移中具有治疗意义的靶点。这些目标的成功完成将为 为未来治疗转移性乳腺癌的新机制的治疗目标奠定基础。 我是威尔康奈尔/洛克菲勒/斯隆凯特琳三机构医学博士-博士生 项目，在洛克菲勒大学 Sohail Tavazoie 博士的实验室中完成了拟议的目标。塔瓦佐伊博士， 以及我的委员会成员博士。 Charles Rice、Ping Chi 和 Kivanc Birsoy 提供了强有力的科学依据 专业知识、指导和资源，以确保项目的成功完成。我们一起采取了 确保我的培训计划也允许进行必要的口头和书面沟通的步骤 技能以及指导机会。我的临床导师，博士。帕梅拉·查尼 (Pamela Charney) 和平池 (Ping Chi) 将提供 在我完成医学培训和选择的过渡过程中，关于保持临床敏锐度的重要建议 内科住院医师研究项目。圆满完成本次拟定计划 一流的培训环境将增强我的 RNA 生物学基础知识，并教会我以下方面的技能： 癌症生物学和小鼠模型，使我坚定地走上成为一名医学科学家的道路， 学术医疗中心的独立调查员。