Connecting Genomic Alterations in Liposarcomas with Drug Responses and Identification of New Therapeutic Approaches

项目摘要 2015年，美国约有12，000人将患上软组织肉瘤。在成人中，脂肪肉瘤 (LPS)是最常见的软组织肉瘤之一。虽然罕见，LPS肿瘤通常非常 发病率和死亡率高。对常规化疗药物的耐药性和缺乏 有效的靶向治疗迫切需要为患有这种疾病的患者开发新的治疗选择。 毁灭性的疾病我们和其他团体最近的大规模分析提供了一个 全面 LPS的基因组景观。尽管有新的见解，这些研究，在大多数情况下， 翻译成 明确的治疗策略。一个挑战是确定个体基因 - 毒品 关系 影响药物反应 第二个挑战是LPS中鉴定的许多遗传改变， 还没有与合适的靶向治疗配对。合成杀伤力是一种有吸引力的方法， 治疗靶向缺乏其他明显的可操作的遗传弱点的癌细胞。大规模 癌症细胞系筛选有助于揭示将遗传决定因素与药物联系起来的合成致命相互作用 然而，由于大多数癌细胞系的复杂性，区分单个突变的相关性 药物的影响仍然很困难。在目前的提案中，我们将采用创新的方法来解决 这些挑战。我们的目标是将特定LPS相关的遗传改变与对 多种临床相关化合物。因此，我们的研究将开始，以弥合之间的差距过多的 我们和其他人在LPS基因组和临床实施中发现的遗传改变。我们将使用一个 基于部分转化的间充质干细胞（MSC）的新筛选策略，我们将遗传 工程化以表达一系列特异性、复发性LPS基因型改变。由此产生的同基因组 克隆和亲本对照细胞将进行高通量小分子和siRNA筛选 （与波特兰俄勒冈州大学的J.泰纳博士和拉霍亚的桑福德·伯纳姆合作）。结果将 在LPS细胞系和动物模型中进行验证和功能研究。此外，我们将执行一项 尖端的全基因组CRISPR筛选，以识别与我们认为是最重要的化合物协同作用的药物。 将在我们的初始屏幕中识别。影响：我们将对药物敏感性进行综合分析 一组同基因MSC克隆中的通路改变和基因型特异性依赖性;结果可能是 发现以前意想不到的基因-药物相互作用，使我们能够提名可行的治疗药物， 靶向并建议联合治疗以减轻耐药性。最终，我们的研究将有助于指导 新的临床试验的各种研究，并帮助开发新的基因型- , FDA批准的药物， 目前缺乏令人满意的治疗选择的LPS患者亚群的定向疗法。

PROJECT SUMMARY Approximately 12,000 individuals in the USA will develop soft tissue sarcoma in 2015. In adults, liposarcoma (LPS) is one of the most common types of soft tissue sarcoma. Though rare, LPS tumors are often very aggressive with high morbidity and mortality rates. Resistance to conventional chemotherapeutics and lack of effective targeted therapies create a critical need to develop novel treatment options for patients with this devastating disease. Recent large-scale analyses by us and other groups have provided a comprehensive view of the genomic landscape of LPS. Despite the new insights, these studies, for the most part have not translated into definitive therapeutic strategies. One challenge is determining how individual gene -drug relationships affect drug responses ; a second challenge is that many of the genetic alterations identified in LPS have not been paired to a suitable targeted therapy. Synthetic lethality is an attractive approach for selective therapeutic targeting of cancer cells lacking otherwise obvious actionable genetic vulnerabilities. Large-scale cancer cell line screens helped uncover synthetic lethal interactions that link genetic determinants to drug responses; yet due to the complexity of most cancer cell lines, distinguishing the relevance of single mutations to the impact of a drug remains difficult. In the current proposal we will use an innovative approach to address these challenges. Our aim is to link specific LPS associated genetic alterations with sensitivities to multiple clinically relevant compounds. Thus, our study will begin to bridge the gap between the plethora of genetic alterations that we and others identified in LPS genomes and clinical implementation. We will use a novel screening strategy based on partly transformed mesenchymal stem cells (MSC) which we will genetically engineer to express a series of specific, recurrent LPS genotypic alterations. The resulting panel of isogenic clones and parental control cells will be subjected to high-throughput small molecule and siRNA screens (Collaboration with Dr. J. Tyner, Oregon University, Portland, and Sanford Burnham, La Jolla). Results will be validated and functionally investigated in LPS cell lines and animal models. In addition, we will perform a cutting-edge, genome-wide CRISPR screen to identify drugs that synergize with a leading compound that we will identify in our initial screens. Impact: We will conduct an integrative analysis of drug sensitivities p athway alterations and genotype-specific dependencies in a panel of isogenic MSC clones; results are likely to uncover previously unexpected gene-drug interactions that will allow us to nominate actionable therapeutic targets and suggest combinational treatments to mitigate drug resistance. Ultimately, our study will help guide new clinical trials for various investigational and help develop new genotype- , FDA-approved and drugs, directed therapies for subsets of LPS patients currently lacking satisfactory therapeutic options.