Elucidating the role of RNA splicing in the metastatic seeding and outgrowth of aggressive cancers.

阐明 RNA 剪接在侵袭性癌症的转移播种和生长中的作用。

• 批准号: 10665061
• 负责人: Kimberly Parker
• 金额: $ 4.32万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-13 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10665061
• 关键词: Address; Affect; Alternative Splicing; Automobile Driving; Binding; Bioinformatics; Biology; Breast; Breast Cancer Cell; Breast cancer metastasis; Cancer Etiology; Cell Survival; Cells; Cessation of life; Clinical; Clinical Research; Coiled-Coil Domain; Complex; Data; Development; Disease; Disseminated Malignant Neoplasm; Distal; Epigenetic Process; Event; Fibroblasts; Foundations; Future; Genes; Genetic; Goals; Grant; Heterogeneity; Immune Evasion; Immunologic Surveillance; In Vitro; Individual; Integrins; International; Invaded; Investigation; Learning; Ligase; Malignant Neoplasms; Mediating; Mentors; Mentorship; Metastatic breast cancer; Methods; Molecular; Neoplasm Metastasis; Organ; Paper; Patient-Focused Outcomes; Phase; Phenotype; Play; Population; Postdoctoral Fellow; Primary Neoplasm; Process; Proliferating; Protein Isoforms; Proteins; RNA; RNA Processing; RNA Splicing; RNA-targeting therapy; Regulation; Relapse; Research; Research Activity; Research Institute; Research Project Grants; Research Proposals; Role; Scientist; Signal Transduction; Site; T-Lymphocyte; Techniques; Therapeutic; Tissues; Transcript; United States; Untranslated RNA; Validation; Variant; Work; Writing; angiogenesis; cancer cell; cancer stem cell; career; crosslinking and immunoprecipitation sequencing; improved; in vivo; malignant breast neoplasm; meetings; neoplastic cell; novel; novel therapeutic intervention; overexpression; post-doctoral training; potential biomarker; pre-doctoral; preclinical study; programs; response; self-renewal; skills; stem cell biomarkers; student mentoring; symposium; targeted treatment; therapeutic target; tool; transcriptome sequencing; transcriptomics; triple-negative invasive breast carcinoma; tumor growth; tumor initiation; tumor microenvironment; tumorigenic

Project Summary/Abstract Metastasis is the process in which disseminated tumor cells (DTCs) spread to distal tissues and organs, and it is the leading cause of cancer-related deaths. In order to complete the metastatic cascade, DTCs must be able to invade distal sites, initiate tumor growth, proliferate, and promote a tumor microenvironment (TME) to evade immune surveillance and promote angiogenesis. Elucidating novel mechanisms that underlie these metastatic phenotypes in DTCs can identify key drivers of metastasis, as well as potential therapeutic vulnerabilities which can better target metastatic tumors. My dissertation research focuses on elucidating the mechanisms in which the lncRNA BORG (BMP/OP-Responsive Gene) drives breast cancer (BC) metastasis. My studies determined that metastatic BC cells are reliant upon BORG complexing with E3 SUMO ligase TRIM28 to induce breast cancer stem cells (BCSCs) to expand and self-renew both in vitro and in vivo. This work establishes BORG as a novel driver of BCSCs, and therefore increases our understanding of the mechanisms driving the accumulation of these malignant cell populations. My recent preliminary data suggests BORG:TRIM28 complexes downregulate the expression of splicing factors in BC cells, which contributes to BCSC phenotypes. Interestingly, accumulating evidence indicates that dysregulation of RNA splicing, a complex molecular tool that can control cellular phenotypes through transcriptomic regulatory mechanisms, contributes to a variety of tumorigenic phenotypes and even metastatic relapse. Therefore, during the F99 portion of this proposal, I seek to elucidate the mechanism whereby BORG:TRIM28 complexes dysregulate RNA splicing operant in driving metastatic and BCSC phenotypes in BC. To complete this research during my dissertation, I will (i) determine which splicing factors dysregulated by BORG:TRIM28 complexes drive metastatic phenotypes in BC in vitro and in vivo, and (ii) use RNAseq and subsequent validation to determine the specific alternative splicing events affected by a subset of these splicing factors in BORG-expressing cells. This work will identify novel mechanisms of RNA splicing regulation that play key roles in metastatic phenotypes of BC cells. During the K00 phase of this proposal, I plan to work closely with my postdoctoral mentor to elucidate the mechanisms in which RNA splicing contributes to tumor cell crosstalk with the TME to promote immune evasion and subsequent metastatic outgrowth. Specifically, I plan to elucidate (i) how alternative splicing in tumor cells drive changes in the metastatic TME to promote immune evasion, and (ii) how alternative splicing changes in TME cells, such as T cells and fibroblasts, contribute to the metastatic niche and subsequent metastatic outgrowth. The current and future goals of my research are to elucidate mechanisms in which dysregulated RNA splicing contributes to metastatic seeding and outgrowth in aggressive cancers. My predoctoral and postdoctoral research will provide me with expertise in the role of RNA splicing in cancer metastasis, and provide me with essential research and professional skills to launch me into my independent research career at a leading research institute.

项目总结/摘要 转移是播散性肿瘤细胞（DTC）扩散到远端组织和器官的过程， 是癌症相关死亡的主要原因。为了完成转移级联，DTC必须能够 侵入远端部位，启动肿瘤生长，增殖，并促进肿瘤微环境（TME）逃避 免疫监视和促进血管生成。阐明这些转移性肿瘤的新机制 DTC的表型可以识别转移的关键驱动因素，以及潜在的治疗弱点， 可以更好地靶向转移性肿瘤。我的博士论文研究的重点是阐明机制， 博格（BMP/OP-应答基因）驱动乳腺癌（BC）转移。我的研究表明 转移性BC细胞依赖于博格与E3 SUMO连接酶TRIM 28复合以诱导乳腺癌的发生。 癌症干细胞（BCSC）在体外和体内扩增和自我更新。这项工作将博格确立为 一种新的BCSC驱动因子，因此增加了我们对驱动BCSC积累的机制的理解。 这些恶性细胞群。我最近的初步数据表明博格：TRIM 28复合物 下调BC细胞中剪接因子的表达，这有助于BCSC表型。有趣的是， 越来越多的证据表明，RNA剪接的失调，一个复杂的分子工具，可以控制 细胞表型通过转录调控机制，有助于多种致瘤性 表型甚至转移复发。因此，在本建议的F99部分，我要求澄清 博格：TRIM 28复合物在驱动转移和转移中的RNA剪接操纵子失调的机制， BC中的BCSC表型。为了在我的论文中完成这项研究，我将（i）确定 由博格：TRIM 28复合物失调的因子在体外和体内驱动BC中的转移表型，以及 (ii)使用RNAseq和随后的验证来确定特定的可变剪接事件， 在BORG表达细胞中这些剪接因子的子集。这项工作将确定RNA的新机制 剪接调控在BC细胞的转移表型中起关键作用。在本提案的K 00阶段， 我计划与我的博士后导师密切合作，阐明RNA剪接有助于 肿瘤细胞与TME相互作用以促进免疫逃避和随后的转移性生长。 具体来说，我计划阐明（i）肿瘤细胞中的选择性剪接如何驱动转移性TME的变化， 促进免疫逃避，和（ii）TME细胞，如T细胞和成纤维细胞， 有助于转移小生境和随后的转移生长。我的当前和未来目标 研究旨在阐明RNA剪接失调导致转移性播种的机制， 恶性肿瘤的副产物。我的博士前和博士后研究将为我提供专业知识 RNA剪接在癌症转移中的作用，并为我提供必要的研究和专业技能， 让我在一家领先的研究机构开始我的独立研究生涯。