Molecular understanding of leukemic bone marrow cytokine-Ras signals and metabolic dependence

白血病骨髓细胞因子-Ras 信号和代谢依赖性的分子理解

基本信息

批准号：
10363571
负责人：
JEROEN ROOSE
金额：
$ 38.36万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10363571
关键词：
20 year old Acute Lymphocytic Leukemia Acute T Cell Leukemia Bar Codes Biochemical Biological Bone Marrow Bone Marrow Diseases Bone Marrow Stem Cell CCL21 gene CRISPR interference Cancer Etiology Cell Line Cells Cellular Metabolic Process Cessation of life Characteristics Childhood Childhood Acute Lymphocytic Leukemia Clinic Clinical Combined Modality Therapy DNA Sequence Alteration Data Dependence Development Epithelial Cells FRAP1 gene Flow Cytometry Genetic Goals Grant Guanosine Triphosphate Hematopoiesis Hematopoietic Hematopoietic stem cells Heterogeneity Immune In Vitro KRASG12D Lead Leukemic Cell Malignant Neoplasms Measurement Measures Metabolic Metabolism Methods Mission Modeling Molecular Mus Mutation Nutrient Oncogenic Pathway interactions Patients Persons Phenotype Phosphatidylinositide 3-Kinase Inhibitor Phosphatidylinositols Phosphotransferases Point Mutation Progress Reports Proto-Oncogene Proteins c-akt RALGDS gene Research Research Personnel Resistance Resolution Role Sampling Signal Transduction T-Cell Leukemia T-Lymphocyte Technology Testing Time Tubulin United States National Institutes of Health Vincristine biobank cancer type chemotherapy cytokine design experience fitness genome-wide high dimensionality in vivo in vivo evaluation inhibitor inhibitor therapy innovation insight interest kinase inhibitor leukemia mouse model new combination therapies novel overexpression pre-clinical precision medicine preclinical trial programs small molecule inhibitor stem stem cells synergism

项目摘要

PROJECT SUMMARY/ABSTRACT Childhood Acute Lymphoblastic Leukemia (ALL) is a disease of the bone marrow with expansion of immature cells and treated with non-specific chemotherapy. ALL remains one of the leading causes of cancer death in persons <20 years old. The developmental origin of T cell ALL (T-ALL) is and profoundly impacts the clinic. Genetic mutations pointed aberrant Ras signals with causal role in 50% of T-ALL, but how such aberrant signals impact developmental trajectories in T-ALL is not known. Similarly, it is by and large unknown how to effectively inhibit aberrant Ras signals. During 1R01CA187318 we uncovered a new molecular paradigm in Ras signaling; Oncogenic Ras mutations (KRasG12D) and overexpression of the Ras activator RasGRP1 drive two very distinctive leukemic Ras signals. In the progress report with our new RoLoRiG/Mx1CRE mouse model, we show that these two aberrant Ras signals both induce abnormal hematopoiesis, but with opposing stem cell features. Capitalizing on our quantitative, multiplex flow cytometry platform we determined that Ras and RasGRP1 frequently connect to PI3K (phosphoinositide 3-kinase)-AKT/mTOR signaling in T-ALL. Next, we performed four extensive synthetic lethality screens with PI3K inhibitors to identify effective combination therapy in T-ALL. We confirmed ten predicted combination therapies with small molecule inhibitors and the PI3K inhibitor GDC0941 with tubulin inhibitor Vincristine yields broad synergy in five cancer types and in our preclinical mouse trials in vivo. The screens predict many metabolic targets, which remained unexplored. Our renewal focuses on understanding the connection between aberrant Ras-PI3K signals and cell metabolism with the goal of resolving the developmental origin of T-ALL and testing new combination therapies. We will determine the connections between Ras-PI3K signals and cell metabolism in our panel of 10 T-ALL cell lines by investigating six nutrient transporters (SLCs), identified in our screens (Aim 1). We will obtain mechanistic understanding of the signals, metabolic features, and hematopoietic composition and trajectories, as a function of leukemic Ras-PI3K signals with unprecedented resolution. We will continue efforts on RoLoRiG/Mx1CRE and KRasG12D/Mx1CRE mouse models that display completely opposite phenotypes in bone marrow stem cells (Aim 2). Lastly, we will obtain a comprehensive characterization of developmental-, biochemical- and metabolic- programs in pediatric T-ALL with single cell resolution. We will combine these efforts with preclinical trial to test the in vivo efficacy of PI3Kγ- and δ- inhibition, inhibition of SLC13A2 and SLC25A44, and combinations in our T-ALL/NGS platform (Aim 3). The synergistic aims, metabolic insights, new mouse models, patient sample-NSG pipeline, and innovative, single cell-resolution technology platforms (CyTOF, SCENITH, phospho-flow) will allow us to make significant contributions towards molecular understanding of T-ALL and precision medicine in cancer.

项目概要/摘要儿童急性淋巴细胞白血病 (ALL) 是一种骨髓未成熟细胞扩张的疾病细胞和非特异性化疗治疗仍然是癌症死亡的主要原因之一。 20 岁以下的人群 T 细胞 ALL (T-ALL) 的发育起源对临床产生深远的影响。基因突变表明异常的 Ras 信号与 50% 的 T-ALL 具有因果关系，但这种异常是如何发生的信号对 T-ALL 发育轨迹的影响尚不清楚。同样，大体上也不知道如何有效抑制异常Ras信号。在 1R01CA187318 期间，我们发现了致癌 Ras 信号传导的新分子范例；突变 (KRasG12D) 和 Ras 激活剂 RasGRP1 的过度表达导致两种非常独特的白血病在我们新的 RoLoRiG/Mx1CRE 小鼠模型的进度报告中，我们显示了这两个信号。异常的 Ras 信号都会导致造血异常，但具有相反的干细胞特征。在我们的定量、多重流式细胞术平台上，我们确定 Ras 和 RasGRP1 经常连接到 T-ALL 中的 PI3K（磷酸肌醇 3 激酶）-AKT/mTOR 信号传导接下来，我们进行了四次。使用 PI3K 抑制剂进行广泛的合成致死率筛选，以确定 T-ALL 的有效联合疗法。确认了小分子抑制剂和 PI3K 抑制剂 GDC0941 的 10 种预测联合疗法与微管蛋白抑制剂长春新碱在五种癌症类型以及我们的临床前小鼠试验中产生广泛的协同作用屏幕预测了许多尚未探索的代谢目标。我们的更新重点是了解异常 Ras-PI3K 信号与细胞之间的联系代谢，目标是解决 T-ALL 的发育起源并测试新的组合我们将在 10 人小组中确定 Ras-PI3K 信号与细胞代谢之间的联系。我们将通过研究筛选中确定的六种营养转运蛋白 (SLC) 来研究 T-ALL 细胞系（目标 1）。获得对信号、代谢特征和造血成分的机制理解，我们将继续努力，以前所未有的分辨率研究白血病 Ras-PI3K 信号的轨迹。在 RoLoRiG/Mx1CRE 和 KRasG12D/Mx1CRE 小鼠模型上显示完全相反的表型最后，我们将获得发育性骨髓干细胞的全面表征。我们将把这些项目结合起来，用于儿科 T-ALL 的生化和代谢项目。努力进行临床前试验，测试 PI3Kγ- 和 δ- 抑制、SLC13A2 抑制和 SLC25A44，以及我们的 T-ALL/NGS 平台中的组合（目标 3）。新的小鼠模型、患者样本-NSG 管道以及创新的单细胞分辨率技术平台（CyTOF、SCENITH、phosph-flow）将使我们能够在分子生物学领域做出重大贡献了解 T-ALL 和癌症精准医学。