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

项目总结/摘要 乳腺癌转移是美国女性死亡的主要原因。尽管在治疗方面取得了进步 对于早期肿瘤患者，转移性疾病患者的预后仍然很差。以前的工作有 证明了转移RNA（tRNAs）作为乳腺癌转移进展驱动因素的直接作用， 差异丰度导致细胞蛋白质组景观的变化。我假设氨酰化状态， 如tRNA是否带有氨基酸，以及必需的氨酰tRNA合成酶（阿尔斯） 负责tRNA充电，也将在乳腺癌转移进展中发挥作用。为了验证这一 假设，在不同转移潜能的细胞中分析了带电的tRNA，这确定了关键的 在具有较高转移潜能的细胞中亮氨酸tRNA充电的减少。初步异种移植和同基因 小鼠转移研究进一步鉴定了负责装载亮氨酸tRNA的亮氨酰阿尔斯（LARS）， 转移抑制因子在这个建议中，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，我将进一步描述LARS操纵的影响（1）通过击倒， 在异种移植和同基因癌症小鼠模型中的过表达，和（2）通过在小鼠中的基因敲除 乳腺癌的模型在目标2中，我将研究（1）转移抑制的细胞表型， 体外试验和（2）带电的tRNA介导的转移抑制的机制，通过下游 RNA测序分析和核糖体分析。这些拟议的研究将填补文献中的空白：尽管 tRNA在癌症进展中起作用，与其生物发生相关的分子尚未被鉴定为 治疗上有意义的靶点。这些目标的成功实现将为 为具有新机制的未来治疗靶点奠定了基础，用于治疗转移性乳腺癌。 我是威尔康奈尔/洛克菲勒/斯隆凯特琳三机构MD-PhD的MD-PhD学生 计划，完成了洛克菲勒大学Sohail Tavazoie博士实验室提出的目标。塔瓦佐医生， 沿着我的委员会成员查尔斯·赖斯博士、平齐博士和基万茨·伯索伊博士， 专业知识、指导和资源，以确保项目的成功完成。我们一起 确保我的培训计划也允许发展必要的口头和书面沟通的步骤 技能和指导机会。我的临床导师帕梅拉·查尼博士和平智博士将提供 在我完成医学培训和选择的过程中， 研究跟踪住院医师计划在内科。成功完成本计划 一流的培训环境将提高我对RNA生物学的基础知识，并教会我以下技能： 癌症生物学和小鼠建模，使我坚定地走上成为一名医生科学家的道路， 学术医学中心的独立调查员

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.