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

基本信息

批准号：
10296685
负责人：
Maria Christina Passarelli
金额：
$ 5.18万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
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/10296685
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 Prognosis 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 overexpression programs ribosome profiling skills small hairpin RNA therapeutic target transcriptome sequencing tumor tumor initiation tumor progression

项目摘要

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.

项目摘要/摘要乳腺癌转移是美国妇女死亡率的主要原因。尽管治疗进展对于患有早期肿瘤的患者，对于转移性疾病的患者，预后仍然很差。先前的工作有通过差异丰度导致细胞蛋白质组学景观的变化。我假设氨基酰化状态，例如，是否将tRNA充满氨基酸，以及必要的氨基酰基tRNA合成酶（AARS）负责tRNA充电的负责也将在乳腺癌转移性进展中发挥作用。测试这个假设，带电的TRNA在不同的转移电位的细胞之间进行了介绍，该细胞确定了钥匙亮氨酸tRNA的减少在具有较高转移性电位的细胞中。初步异种移植和同步小鼠转移研究进一步鉴定出负责为亮氨酸TRNA充电的亮氨酸AARS（LARS）作为A 转移抑制剂。在此提案中，我将研究并进一步描述LARS在癌症中的作用转移。在AIM 1中，我将进一步描述Lars操纵（1）的影响（1）异种移植物和促癌小鼠模型中的过表达，（2）通过小鼠的遗传敲除乳腺癌的模型。在AIM 2中，我将研究（1）通过体外测定和（2）通过下游带电的TRNA介导的转移的机理 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：不过 TRNA在癌症进展中起作用，与其生物发生相关的分子尚未确定为癌症转移的治疗靶标有意义。这些目标的成功完成将是未来治疗靶标的未来治疗靶标的基础工作，用于治疗转移性乳腺癌。我是Weill Cornell/Rockefeller/Sloan Kettering三机构MD-PHD的MD-PHD学生计划，完成洛克菲勒大学Sohail Tavazoie博士的实验室中提议的目标。 Tavazoie博士，以及我的委员会成员Drs。查尔斯·赖斯（Charles Rice），Ping Chi和Kivanc Birsoy提供了强大的科学专业知识，指导和资源，以确保成功完成项目。我们在一起确保我的培训计划的步骤还允许发展必要的口头和书面交流技能和指导机会。我的临床导师，博士。 Pamela Charney和Ping Chi将提供当我过渡到完成医学培训并选择时，关于保持临床敏锐的关键建议内科研究中的研究轨道居住计划。在此成功完成了拟议计划出色的培训环境将增强我对RNA生物学的基础知识，并教我技能癌症生物学和小鼠建模，使我坚定地成为医师科学家的道路学术医学中心的独立研究员。