New models and therapeutic approaches in alveolar rhabdomyosarcoma

项目总结(30行) 肺泡型横纹肌肉瘤(ARMS)是一种常见的侵袭性肌肉癌，影响数百例 美国每年都有儿童。ARM具有致癌的染色体易位 在PAX基因和叉头转录因子(FOXO1)之间。然而，PAX3/7的通路- Foxo1转录因子调节肿瘤细胞的起源，以及出现的治疗脆弱性 手臂细胞仍然不清楚，这主要是因为缺乏准确概括相同情况的精确动物模型。 在人类手臂中发现的易位融合事件。我们工作的长期目标是从治疗角度揭示 推动武器增长的相关途径。这个应用程序的总体目标是开发斑马鱼 武器模型，以确定可在临床上利用的治疗脆弱性。我们的中心假设是 在PAX3-FOXO1臂中，较差的生存结果反映在起源细胞的差异上， 分子定义的肿瘤增殖细胞(TPC)的数量和转移的易感性。可行性 我们的方法得到了我们使用斑马鱼模拟各种人类癌症的工作的支持，最近 优化Crispr/Cas9方法以使用Cre/Lox和我们的 成功的高含量成像筛选方法识别FDA批准的具有抑制效果的药物 其他RMS亚型的生长。我们研究理由是很少有好的实验动物 精确地模拟人类手臂的潜在遗传学的模型，使我们无法定义 特定的致癌融合会推动癌症的生长。这项工作意义重大，因为它将揭示 解释PAX3/7-FOXO1臂临床表现差异的细胞机制 确定治疗手臂的新疗法，并为易位癌症提供新的建模方法，所有这些 在PA-16-251中表达了由癌症登月倡议支持的目标。目标1将描述 用创新的斑马鱼模型比较PAX3/7-FOXO1诱导的手臂的差异，验证我们的假设 聚合臂在增殖和细胞来源(S)方面有内在的差异。AIM 2将评估PAX3/7-FRKH 对于调节转移和肿瘤增殖潜力的不同作用，检验我们的假设 Pax3-FOXO1臂的临床表现较差，因为它们具有较高的TPC数量和转移能力。 AIM 3将使用创新的高内容成像屏幕来识别FDA批准的杀死TPC和 抑制人类手臂的生长。关于结果，我们的研究将发展出急需的精确度。 武器的动物模型并确定PAX3/7-FOXO1武器的主要差异，包括可能的差异 原始细胞、TPC和转移能力，解释了为什么PAX3-FOXO1臂表现更差 从临床上看。我们的工作还将确定新的治疗方法来杀死人类患者来源的异种移植物(PDX)中的TPC。 这项工作预计将产生积极的翻译影响，通过展示临床前的疗效 靶向人类手臂中的TPC，并确定治疗这种毁灭性癌症的新疗法。

PROJECT SUMMARY (30 lines) Alveolar rhabdomyosarcoma (ARMS) is a common and aggressive muscle cancer that affects hundreds of children annually in the United States. ARMS are pathognomonic with oncogenic chromosomal translocations between the PAX genes and the fork-head transcription factor (FOXO1). Yet, the pathways that the PAX3/7- FOXO1 transcription factors modulate, the tumor cells of origin, and the therapeutic vulnerabilities that arise in ARMS cells is still unclear, largely due to lack of precision animal models that accurately recapitulate the same translocation fusion events found in human ARMS. The long-term goal of our work is to uncover therapeutically relevant pathways that drive ARMS growth. The overall objective of this application is to develop zebrafish models of ARMS to identify therapeutic vulnerabilities that can be exploited clinically. Our central hypothesis is that worse survival outcomes in PAX3-FOXO1+ ARMS are reflected in differences in cells of origin, elevated numbers of molecular defined tumor-propagating cells (TPCs), and predisposition to metastasis. The feasibility of our approach is supported by our work in using zebrafish to model a wide range of human cancers, recent optimization of Crispr/CAS9 approaches to create patient-specific translocations using CRE/Lox, and our successful high-content imaging screening approach to identify FDA approved drugs with efficacy in curbing growth of other RMS subtypes. The rationale for our research is that there are few good experimental animal models that accurately mimic the underlying genetics of human ARMS, obviating our ability to define how specific oncogenic fusions drive cancer growth. This work is significant because it will uncover divergent cellular mechanisms that account for differences in the clinical manifestation of PAX3/7-FOXO1+ ARMS, identify new therapies to treat ARMS, and provide new modeling approaches for translocation+ cancers, all expressed goals outlined in PA-16-251 supported by the Cancer Moonshot Initiative. Aim 1 will characterize differences in PAX3/7-FOXO1-induced ARMS using innovative zebrafish models, testing our hypothesis that fusion+ ARMS have inherent differences in proliferation and cell(s)-of-origin. Aim 2 will assess PAX3/7-FRKH for differential effects on modulating metastasis and tumor propagating potential, testing our hypothesis that PAX3-FOXO1+ ARMS do worse clinically because they have elevated TPC numbers and metastatic capacity. Aim 3 will use an innovative high-content imaging screen to identify FDA-approved drugs that kill TPCs and suppress growth of human ARMS. With respect to outcomes, our research will develop much-needed precision animals models of ARMS and identify key differences in PAX3/7-FOXO1+ ARMS including likely differences in cell-of-origin, TPCs, and metastatic capacity, providing explanation of why PAX3-FOXO1+ ARMS do worse clinically. Our work will also identify novel therapies to kill TPCs in human patient derived xenografts (PDX). This work is expected to have a positive translational impact by demonstrating the pre-clinical efficacy of targeting TPCs in human ARMS and identifying new therapies for the treatment of this devastating cancer.