Defining the biological consequences of somatic mutations in high risk AML using integrative single cell approaches

项目摘要 急性髓系白血病（AML）是一种侵袭性血癌，患者预后极差。 骨髓肿瘤的系统测序研究已经确定了AML中的复发突变， 为靶向治疗（如IDH 1/2和FLT 3）和分子指导的临床治疗的发展提供了机会。 护理（诊断，风险分层）。尽管最近取得了这些进展，但这些突变如何特别驱动 发病机制和疾病进展以及确定治疗反应的认识较少。定义 这些关系受到AML基因组遗传和克隆多样性的挑战。这项建议旨在 研究获得性突变在两种互补背景下的生物学和临床作用：1）IDH 1/2突变 诊断时和IDH抑制剂治疗（靶向治疗）下的AML; 2）TP 53突变型克隆造血 在实体瘤的细胞毒性治疗下，AML的转化（早期检测和干预）。 为了解决这个问题，我们建立了一种方法来获得单细胞基因型和基因表达。我们 表明我们可以将单个细胞分配到亚克隆中，并在AML中提供亚克隆特异性基因表达谱。 IDH 1和IDH 2突变的靶向抑制剂最近获得FDA批准用于复发性/难治性AML， 然而，反应仍然很不稳定。在这些临床试验中，信号传导亚克隆突变的患者 或剪接基因的人更有可能是无反应者。第一个具体目标是界定后天的作用 诊断时和使用整合性单细胞的IDH抑制剂治疗下IDH 1/2突变型AML的突变 接近。我们假设信号传导和剪接突变的生物效应子驱动亚克隆 介导抗性的基因表达谱，不同于对烟草敏感的显性克隆。 我们的初步数据确定了克隆性造血中TP 53的突变与随后的 获得等位基因失衡和在实体瘤化疗下转化为AML。第二 具体目的是研究TP 53突变体克隆性造血向治疗相关转化的机制 使用整合性单细胞方法的髓样肿瘤。我们将检验TP 53突变 为等位基因失衡提供肥沃的土壤，这些畸变共同推动AML的进展。 总之，这些目标告知个体突变如何在疾病期间促进骨髓发病机制 开始，进展，并在治疗的选择性压力下。这将有助于了解AML疾病 生物学、临床反应以及新的早期检测和治疗策略的开发。 Elli Papaemmanuil博士和Ross Levine博士，在功能和临床后果方面具有专业知识， 骨髓恶性肿瘤中的体细胞突变作为本申请的申办者。关于Gerstner Sloan Kettering 研究生院为申请人提供最佳的培训环境。此次培训将推动 申请人的目标是成为癌症生物学和基因组学前沿的主要研究者。

Project Summary Acute Myeloid Leukemia (AML) is an aggressive blood cancer with exceedingly poor patient outcomes. Systematic sequencing studies in myeloid neoplasms have identified the recurrent mutations in AML, raising opportunities for the development of targeted therapeutics (e.g. IDH1/2 and FLT3) and molecularly guided clinical care (diagnosis, risk stratification). Despite these recent advances, how these mutations specifically drive pathogenesis and disease progression and define treatment responses is less well understood. Definition of these relationships has been challenged by the genetic and clonal diversity of AML genomes. This proposal aims to study the biological and clinical role of acquired mutations in two complementary contexts: 1) IDH1/2 mutant AML at diagnosis and under IDH inhibitor therapy (targeted therapeutics); 2) TP53 mutant clonal hematopoiesis transformation to AML under cytotoxic therapy for solid tumors (early detection and intervention). To address this question, we established a method to derive single cell genotype and gene expression. We show we can assign single cells to subclones and deliver subclone-specific gene expression profiles in AML. Targeted inhibitors for IDH1 and IDH2 mutations recently attained FDA approval in relapsed/refractory AML, however response remains highly variable. In these clinical trials, patients with subclonal mutations in signaling or splicing genes were more likely to be non-responders. The first specific aim is to define the role of acquired mutations in IDH1/2-mutant AML at diagnosis and under IDH inhibitor therapy using integrative single cell approaches. We hypothesize that the biological effectors of signaling and splicing mutations drive subclonal gene expression profiles mediating resistance, distinct from the inhibitor-sensitive dominant clone. Our preliminary data identifies mutations in TP53 at clonal hematopoiesis are associated with subsequent acquisition of allelic imbalances and transformation to AML under chemotherapy for solid tumors. The second specific aim is to study mechanisms of TP53 mutant clonal hematopoiesis transformation to therapy-related myeloid neoplasm using integrative single cell approaches. We will test the hypothesis that TP53 mutations provide fertile ground for allelic imbalances and these aberrations together drive progression to AML. Together, these aims inform how individual mutations contribute to myeloid pathogenesis during disease initiation, progression, and under the selective pressure of treatment. This will provide insight into AML disease biology, clinical response, and the development of new early detection and therapeutic strategies. Dr. Elli Papaemmanuil and Dr. Ross Levine, with expertise in the functional and clinical consequences of somatic mutations in myeloid malignancies, serve as Sponsors of this application. The Gerstner Sloan Kettering Graduate School provides the optimal training environment for the applicant. This training will advance the applicant’s goal to become a principal investigator on the frontier of cancer biology and genomics.