Leveraging in vivo models to identify epigenetic vulnerabilities in leukemia

利用体内模型识别白血病的表观遗传脆弱性

• 批准号: 8878386
• 负责人: ANDREW L KUNG
• 金额: $ 17.4万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8878386
• 关键词: Acute T Cell Leukemia; Acute leukemia; Address; Biological Assay; Cancer Biology; Cancer Model; Cell Culture Techniques; Cell Line; Chemicals; Chimeric Proteins; Clinic; Clinical Trials; Complex; DNA Methylation; Dependency; Derivation procedure; Development; Disease; Drug Targeting; Emerging Technologies; Enzymes; Epigenetic Process; Explosion; Failure; Frequencies; Gene Expression; Genes; Genetic; Genetic Screening; Genetically Engineered Mouse; Goals; Growth; Histone Deacetylase Inhibitor; Human; In Vitro; Individual; Induction of Apoptosis; Laboratories; Libraries; Link; Malignant Neoplasms; Metabolism; Methods; Mixed-Lineage Leukemia; Modeling; Modification; Mus; Mutate; Mutation; Oncogenes; Patients; Phosphotransferases; Play; Population; Predictive Value; Proteins; Publications; Role; Terminal Disease; Therapeutic; Translations; Validation; Xenograft Model; Xenograft procedure; base; cancer therapy; cell growth; clinically relevant; drug development; drug discovery; drug testing; epigenetic regulation; epigenome; genetic regulatory protein; genome-wide; improved; in vitro Model; in vivo; in vivo Model; inhibitor/antagonist; innovative technologies; interest; leukemia; monolayer; mouse model; next generation sequencing; outcome forecast; predictive modeling; pressure; public health relevance; screening; small hairpin RNA; subcutaneous; success; targeted treatment; therapeutic target; tumorigenic

 DESCRIPTION (provided by applicant): Mouse models of cancer are a critical link between basic laboratory discovery and translation to human clinical trials. For target validation, in vivo models are often the ultimate arbiters of validity (e.g., for publication), and in drug development no new chemical entity advances without evidence of efficacy utilizing in vivo models. Although the predictive value of mouse cancer models is subject to debate, in vivo models do recapitulate many aspects of cancer biology that are absent from in vitro cell line models under non-physiological cell culture conditions. For pragmatic reasons, however, most large-scale target identification efforts strictly utilize cell line-based screening methods despite the clear limitatons of in vitro models. We hypothesize that incorporating in vivo models into primary target identification screens will facilitate the identification of clinically relevant targets, and may identify targets that would not be evident using only in vitro cell line screening approaches. Given the explosion of interest in epigenetic therapies, in this proposal we will use a pooled shRNA approach to comprehensively identify epigenetic vulnerabilities in T-cell acute lymphoblastic leukemia (T- ALL) and mixed lineage leukemia (MLL). Using a library of 3,040 shRNAs targeting 449 known and putative epigenetic regulatory proteins, we will simultaneously assess for dependencies using in vitro cell line models and in vivo orthotopic leukemia models. In the first Aim, we will surmount one of the major impediments to in vivo screening by deriving highly leukemogenic sub-lines representing T-ALL and MLL. There is an unmet need for better therapies for these poor prognosis leukemia subclasses. In the second Aim, we will use pooled shRNA assays, performed simultaneously in vitro and in vivo, to identify epigenetic vulnerabilities in these two subclasses of leukemia. Finally, in the third Aim, we will fully validte critical vulnerabilities, including achieving in vivo proof-of-concept for therapeutic targeting. Together, these studies utilize a convergence of innovative and emerging technologies to identify epigenetic vulnerabilities in leukemia. Success in these studies would establish a new paradigm for target discovery leveraging in vivo models at the inception of discovery campaigns. In addition to rapid elimination of false positive hits, this approach may identify new target classes that are opaque to standard in vitro screening methods.

 描述（由申请人提供）：小鼠癌症模型是基础实验室发现和转化为人体临床试验之间的关键环节。用于体内靶点验证 模型通常是有效性的最终仲裁者（例如，出版），以及药物开发 在没有利用体内模型的有效性证据的情况下，没有新的化学实体进展。尽管小鼠癌症模型的预测价值受到争议，但体内模型确实概括了在非生理细胞培养条件下体外细胞系模型中不存在的癌症生物学的许多方面。然而，出于实用的原因，尽管体外模型有明显的局限性，但大多数大规模的靶点鉴定工作严格利用基于细胞系的筛选方法。 我们假设，将体内模型纳入初级靶标鉴定筛选将有助于临床相关靶标的鉴定，并且可以鉴定仅使用体外细胞系筛选方法不明显的靶标。鉴于对表观遗传疗法的兴趣激增，在本提案中，我们将使用汇集的shRNA方法来全面鉴定T细胞急性淋巴细胞白血病（T-ALL）和混合谱系白血病（MLL）中的表观遗传脆弱性。使用靶向449个已知和推定的表观遗传调节蛋白的3，040个shRNA文库，我们将使用体外细胞系模型和体内原位白血病模型同时评估依赖性。在第一个目标中，我们将通过衍生代表T-ALL和MLL的高度致白血病的亚系来克服体内筛选的主要障碍之一。对这些预后不良的白血病亚类的更好疗法存在未满足的需求。在第二个目标中，我们将使用合并的shRNA检测，同时在体外和体内进行，以确定这两个白血病亚类的表观遗传脆弱性。最后，在第三个目标中，我们将充分验证关键漏洞，包括实现治疗靶向的体内概念验证。 总之，这些研究利用创新和新兴技术的融合来识别白血病的表观遗传脆弱性。这些研究的成功将在发现活动开始时利用体内模型为目标发现建立新的范式。除了快速消除假阳性命中，这种方法可以识别新的目标类别是不透明的标准体外筛选方法。