In Vivo Testing of Novel Drug Combinations for Pediatric Soft Tissue Sarcomas

项目摘要 尽管我们对儿科软组织肉瘤病因的了解取得了进展 (STS)这些疾病的总体生存率在过去20年中没有显著改善。为 患有复发性疾病的儿童，存活率低于30%，长期存活者的存活率增加。 与他们接受的治疗相关的疾病负担。因此，我们的目标是 研究小组的目的是通过整合基本的， 转化和临床研究。在过去的10年里，我们已经同意STS和其他固体 肿瘤患者捐献组织原位植入免疫功能低下的小鼠体内， 原位患者来源的异种移植物（O-PDX）。我们的O-PDX已用于体外高- 使用标准化的临床前I、II、III期范例进行通量药物筛选和体内测试。 横纹肌肉瘤（RMS）是儿童中最常见的STS，基因组研究表明， 罕见的克隆相关细胞亚群可以在治疗后存活并导致疾病复发。 随后使用转录组学、表观遗传学和蛋白质组学数据进行的综合分析表明，RMS 肿瘤保留其发育起源的谱系特异性转录和表观遗传标记。 最近，单细胞和单核RNA-seq（sc/snRNA-seq）和体内谱系追踪（line-tracing）被用于研究细胞内的RNA序列。 表明克隆细胞可以通过正常的发育程序转变。确实，单身-- 细胞ATAC-seq证明，细胞和发育阶段特异性超级增强子活性是 与基因表达的克隆性变化相关。化疗消除了 增殖的肿瘤细胞群，和存活的休眠肿瘤细胞迅速扩大和重新 建立他们的发展层次，这导致疾病复发。这是一个突出的例子 STS中复杂的细胞内在和外在信号传导以及STS与细胞内信号传导之间的复杂联系。 儿童癌症的发展和致癌途径。在本提案中，我们将在体内进行 每年使用我们的STS O-PDX模型测试8-10种药物。最引人注目的途径是 发育激酶途径（Aim 1）、细胞应激途径（Aim 2）和G2/M细胞周期检查点 (Aim 3）。新型药物组合将被测试，以及那些包括传统的 化疗作为标准治疗特别强调的是， 通过减少疾病复发来提高生存率。

PROJECT SUMMARY Despite the advances made in our understanding of the etiology of pediatric soft tissue sarcomas (STS), the overall survival of those diseases has not significantly improved in over 2 decades. For children with recurrent disease, survival is below 30%, and long-term survivors have an increased burden of disease associated with the curative therapies they received. Therefore, the goal of our research team is to improve the survival and quality of life of children with STS by integrating basic, translational, and clinical research. For the past 10 years, we have consented STS and other solid tumor patients to donate tissue for orthotopic implantation into immunocompromised mice to develop orthotopic patient derived xenografts (O-PDXs). Our O-PDXs have been used for ex vivo high- throughput drug screening and in vivo testing using a standardized preclinical phase I, II, III paradigm. Rhabdomyosarcoma (RMS) is the most common STS in children and genomic studies have shown that rare subsets of clonally related cells can survive treatment and contribute to disease recurrence. Subsequent integrated analyses using transcriptomic, epigenetic and proteomic data showed that RMS tumors retain lineage-specific transcriptional and epigenetic signatures of their developmental origins. More recently, single cell and single nucleus RNA-seq (sc/snRNA-seq) and in vivo lineage-tracing showed that clones of cells can transition through their normal developmental programs. Indeed, single- cell ATAC-seq demonstrated that the cell- and developmental stage–specific super-enhancer activity is correlated with those clonal changes in gene expression. Chemotherapy eliminates the most proliferative tumor cell populations, and the surviving dormant tumor cells rapidly expand and re- establish their developmental hierarchy, which leads to disease recurrence. This is a striking example of the complex cell-intrinsic and -extrinsic signaling within STS and the intricate connection between developmental and oncogenic pathways in childhood cancer. In this proposal, we will perform in vivo testing for 8-10 drugs per year using our STS O-PDX models. The most compelling pathways are developmental kinase pathways (Aim 1), cell stress pathways (Aim 2) and G2/M cell cycle checkpoints (Aim 3). Novel drug combinations will be tested as well as those that include conventional chemotherapy for standard of care. Particular emphasis will be placed on eliminating all the clones in the tumor to improve survival by reducing disease recurrence.