DISSECTING THE ROLES OF FAT1 AND NEAT1 IN SOFT TISSUE SARCOMA DEVELOPMENT AND METASTASIS USING NOVEL IN VIVO SARCOMA MODELS

使用新颖的体内肉瘤模型剖析 FAT1 和 NEAT1 在软组织肉瘤发展和转移中的作用

• 批准号: 10697997
• 负责人: Jianguo Huang
• 金额: $ 16.2万
• 依托单位: PROVIDENCE HEALTH & SERVICES - OREGON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-07 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697997
• 关键词: Adult; Biology; CRISPR/Cas technology; Cells; Clinical; Code; Combined Modality Therapy; Complex; Data; Development; Diagnosis; Distant; FAT gene; Foundations; Funding Agency; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Screening; Genetically Engineered Mouse; Genomics; Goals; Grant; High-Throughput RNA Sequencing; Human; Immunohistochemistry; In Vitro; K22 Award; Knock-out; Knowledge; Lung; Malignant Fibrous Histiocytoma; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mesenchymal Cell Neoplasm; Metastatic Neoplasm to the Lung; Methods; Modeling; Modification; Mus; Muscle; Mutate; Mutation; Neoplasm Metastasis; Oncogenes; Oncogenic; Organ; Pathway interactions; Patients; Play; Population; Prognosis; Protein Splicing; Proteins; RNA; RNA Splicing; Research; Resistance; Role; Signal Pathway; Soft tissue sarcoma; Spliced Genes; Survival Rate; Testing; Therapeutic; Tissue Microarray; Translating; Tumor Suppressor Proteins; Untranslated RNA; Up-Regulation; Wild Type Mouse; chemotherapy; design; genome-wide; human data; improved; improved outcome; in vivo; in vivo Model; inhibitor; insight; mouse model; novel; novel therapeutic intervention; overexpression; pharmacologic; preclinical trial; programs; sarcoma; targeted treatment; therapeutically effective; transcriptome sequencing; treatment strategy; tumor; tumor growth

Soft tissue sarcomas (STSs) are rare heterogeneous mesenchymal tumors that have more than 75 subtypes. STSs are understudied tumors for which there are few established research models and a lack of funding sources. Over decades, there has been little improvement in the therapeutic strategies for STSs, which are often resistant to current therapies and can be frequently fatal as 50% of patients develop metastasis in distant organs. To solve this unmet clinical problem, in vivo models that accurately recapitulate this spectrum of cancers provide a unique and effective platform for studying sarcoma biology and preclinical trials before novel therapeutic strategies translate to limited population of sarcoma patients. However, there are very few in vivo sarcoma models available because the tumor suppressor and oncogenic drivers for sarcoma development and metastasis remain unknown. Therefore, I performed genome-wide in vitro genetic screens and direct in vivo CRISPR/Cas9 knockout screens in wild type mice to identify driver genes whose mutation is required for sarcoma initiation. From these screens, I generate a novel in vivo sarcoma model driven by the mutation of Fat1 which is frequently mutated in human STSs. This is a de novo in vivo model that recapitulates a subset of human STSs and, to our knowledge, the first determination that Fat1 is a potent tumor suppressor in human STSs. Furthermore, using in vivo sarcoma models and high throughput RNA sequencing, I also identified the long non-coding RNA (lncRNA) Neat1 as an oncogenic driver for sarcoma metastasis. This K22 award will allow me to build my own research platform to further characterize the critical signaling pathways and target genes in sarcoma development and metastasis using these unique in vivo sarcoma models. In Specific Aim 1, we will dissect the mechanism by which the Hippo pathways and their effectors Yap1/Taz drive sarcomas through the mutation of Fat1. In addition, we will use my novel in vivo sarcoma models to test and optimize the best combination treatment strategies that suppress sarcoma tumor growth. In Specific Aim 2, we will determine the mechanisms by which lncRNA Neat1 drives sarcoma metastasis. My preliminary results suggest that RNA splicing regulating genes, such as Khsrp, interact with Neat1 and promote sarcoma metastasis. We will use my unique in vivo sarcoma models to dissect the mechanisms governing sarcoma metastasis and the implications of these genes for targeted therapies in treating metastatic sarcoma patients. In conclusion, completion of this proposal will determine the functional consequences of expression of the coding gene Fat1 and the non-coding gene Neat1 in sarcoma development and metastasis and provide novel candidate pathways and genes for designing effective targeted therapies to improve outcomes for sarcoma patients.

软组织肉瘤 (STS) 是罕见的异质性间质肿瘤，有超过 75 种亚型。 STS 是 尚未得到充分研究的肿瘤，几乎没有建立的研究模型并且缺乏资金来源。几十年来， STS 的治疗策略几乎没有任何改进，STS 常常对当前的疗法产生耐药性 由于 50% 的患者发生远处器官转移，因此通常可能致命。为了解决这个未满足的临床问题， 准确概括这一系列癌症的体内模型为研究提供了独特而有效的平台 新的治疗策略转化为有限的肉瘤人群之前的肉瘤生物学和临床前试验 患者。然而，由于肿瘤抑制因子和致癌基因，可用的体内肉瘤模型非常少。 肉瘤发展和转移的驱动因素仍然未知。因此，我进行了全基因组体外遗传 在野生型小鼠中进行筛选和直接体内 CRISPR/Cas9 敲除筛选，以鉴定其突变为的驱动基因 肉瘤发生所需的。从这些屏幕中，我生成了一种由突变驱动的新型体内肉瘤模型 Fat1 在人类 STS 中经常发生突变。这是一个从头开始的体内模型，概括了人类的一个子集 STS，据我们所知，首次确定 Fat1 是人类 STS 中的有效肿瘤抑制因子。此外， 使用体内肉瘤模型和高通量 RNA 测序，我还鉴定了长非编码 RNA (lncRNA) Neat1 作为肉瘤转移的致癌驱动因素。这个 K22 奖项将使我能够建立自己的研究平台 利用这些进一步表征肉瘤发展和转移中的关键信号通路和靶基因 独特的体内肉瘤模型。在具体目标 1 中，我们将剖析 Hippo 通路及其作用的机制。 效应子 Yap1/Taz 通过 Fat1 突变驱动肉瘤。另外，我们会用我的小说体内肉瘤 模型来测试和优化抑制肉瘤肿瘤生长的最佳组合治疗策略。具体来说 目标 2，我们将确定 lncRNA Neat1 驱动肉瘤转移的机制。我的初步结果 表明 RNA 剪接调节基因（例如 Khsrp）与 Neat1 相互作用并促进肉瘤转移。我们将 使用我独特的体内肉瘤模型来剖析控制肉瘤转移的机制及其影响 这些基因可用于治疗转移性肉瘤患者的靶向治疗。综上所述，本提案的完成 将决定编码基因 Fat1 和非编码基因 Neat1 表达的功能后果 肉瘤的发展和转移，并为设计有效的靶向药物提供新的候选途径和基因 改善肉瘤患者预后的疗法。