New Paradigms for Targeting Truncal Driver Mutations

针对树干驱动突变的新范例

• 批准号: 9978002
• 负责人: WILLIAM G. KAELIN
• 金额: $ 98.25万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978002
• 关键词: Acute Myelocytic Leukemia; Acute Promyelocytic Leukemia; Address; Affect; Antineoplastic Agents; Arsenic Trioxide; Biological Markers; CRISPR screen; Cancer Model; Cells; Chemicals; Chimeric Proteins; Clear Cell; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Drug resistance; Event; Evolution; Gene Frequency; Genes; Genetic; Knowledge; Lesion; Maintenance; Malignant Neoplasms; Molecular; Monitor; Multiple Myeloma; Mutation; Oncoproteins; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Play; Proteins; Renal carcinoma; Research Personnel; Retinoblastoma Protein; Role; Solid Neoplasm; Technology; Thalidomide; Therapeutic Effect; Trees; Tretinoin; Tumor Suppressor Proteins; Work; base; chemical genetics; deep sequencing; driver mutation; interest; loss of function mutation; lung small cell carcinoma; mouse model; mutant; neoplastic cell; pre-clinical; response; small molecule; therapeutic evaluation; therapeutic target; therapy resistant; transcription factor; tumor; tumor progression; ubiquitin ligase; validation studies; virtual

Summary: Solid tumors generate genetically distinct subclones during their evolution and expansion. Deep sequencing followed by quantification of mutant allele frequencies within a given tumor allows one to infer evolutionary trees consisting of shared early driver (“truncal”) mutations and divergent late driver (“branch”) mutations. This knowledge suggests one should therapeutically target truncal mutations, since they are theoretically shared by all of the cells within a tumor, rather than late mutations. Moreover, it is likely that some mutations that occur late during tumor evolution are only advantageous to (or tolerated by) tumor cells because of the mutations that preceded them. In such cases targeting truncal mutations could have therapeutic effects by unmasking deleterious effects to the tumor cell caused by the late mutations. In fact, virtually every successful targeted cancer drug attacks a genetic event that is known or suspected to be truncal. Combining two drugs that inhibit the same truncal lesion in different ways, such as when combining retinoic acid with arsenic trioxide to inhibit the PML-RAR fusion protein in acute promyelocytic leukemia, should enhance efficacy and reduce therapeutic resistance. The Kaelin Lab has had a longstanding interest in pRB and pVHL tumor suppressor proteins and most recently, in IDH oncoproteins. Mutations affecting these proteins occur as early truncal events in specific cancers such as small cell lung cancer (pRB), clear cell renal cancer (pVHL), and acute myelogenous leukemia (IDH1 and IDH2). The Kaelin Lab has played an important role in demonstrating the roles of pRB loss, pVHL loss, and mutant IDH in tumor maintenance and in identifying their pathogenic downstream targets. This proposal seeks to create new paradigms for targeting truncal mutations, including those currently deemed undruggable (for example, loss of function mutations or mutations encoding proteins without druggable pockets). Loss of function mutations will be addressed by exploiting epistatic relationships and synthetic lethal relationships, using both hypothesis-driven and CRISPR-based screening approaches. The Kaelin Lab recently showed that thalidomide-like drugs redirect the cereblon ubiquitin ligase to degrade the IKF1 and IKF3 transcription factors, which play important roles in myeloma. In the course of this work they developed a technology that allows them to screen for proteins that are destabilized (or stabilized) in response to specific chemical or genetic perturbants, as well as to screen for chemical and genetic perturbants that can destabilize (or stabilize) proteins of interest. The former will be used to identify protein-based biomarkers for monitoring molecular pathways of interest and the latter will be used to look for small molecules/targets capable of destabilizing oncoproteins of interest. They also identified a modular degron with IKZF1/3 that can be used to target heterologous proteins for destruction, which will be incorporated into preclinical target validation studies. Finally, CRISPR-based gene editing will be used to rapidly make mouse models of cancer driven by specific truncal mutations and for testing therapeutic and monitoring strategies emerging from these studies.

概括： 实体瘤在其进化和膨胀过程中产生一般不同的亚克隆。深度测序 其次是给定肿瘤内的突变等位基因频率的数量，可以推断出进化 由共享早期驱动器（“截短”）突变和分歧的晚期驱动器（“分支”）突变组成的树木。这 知识表明，历史上应该针对截短突变，因为理论上是由 肿瘤中的所有细胞，而不是晚期突变。而且，某些突变可能发生 由于突变，肿瘤演化期间晚期仅对（或耐受）肿瘤细胞有利 在他们之前。在这种情况下，针对截短突变的靶向 晚期突变引起的肿瘤细胞的有害作用。实际上，几乎每个成功的目标 癌症药物攻击一种遗传事件，该事件已知或怀疑是截短的。结合两种抑制的药物 以不同的方式具有相同的截短病变 急性临时细胞白血病中的PML-RAR融合蛋白应提高效率并降低治疗 反抗。 Kaelin Lab对PRB和PVHL肿瘤抑制蛋白和 最近，在IDH癌蛋白中。影响这些蛋白质的突变是在特定的早期截断事件中发生的 癌症，例如小细胞肺癌（PRB），透明细胞肾癌（PVHL）和急性骨髓 白血病（IDH1和IDH2）。 Kaelin Lab在展示PRB损失的角色方面发挥了重要作用， PVHL损失和肿瘤维持中的突变体IDH并确定其致病性下游靶标。这 提案旨在创建针对截短突变的新范式，包括当前被视为的突变 不可用 口袋）。功能突变的丧失将通过利用认识关系和合成致命来解决 使用基于假设驱动和基于CRISPR的筛选方法的关系。凯林实验室 最近表明，类似沙利度胺的药物重定向脑泛素连接酶，以降解IKF1和IKF3 转录因子，在骨髓瘤中起重要作用。在这项工作的过程中，他们开发了 使他们能够筛选出对特定的蛋白质筛查（或稳定）的蛋白质的技术 化学或遗传扰动者，以及筛选可能破坏稳定稳定的化学和遗传扰动剂 （或稳定）感兴趣的蛋白质。前者将用于识别基于蛋白质的生物标志物进行监测 感兴趣的分子途径和后者将用于寻找能够的小分子/靶标 不稳定感兴趣的癌蛋白。他们还鉴定了一个带有ikzf1/3的模块化degron 靶标异源蛋白用于破坏，将纳入临床前目标验证研究中。 最后，将使用基于CRISPR的基因编辑来快速使癌症的小鼠模型由特异性驱动 截短突变以及用于测试这些研究的治疗和监测策略。