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：
10545014
负责人：
JEROEN ROOSE
金额：
$ 37.59万
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/10545014
关键词：
20 year old Acute Lymphocytic Leukemia Acute T Cell Leukemia Bar Codes Biochemical Biological Bone Diseases 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 Childhood Precursor T Lymphoblastic 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 Testing Time Tubulin United States National Institutes of Health Vincristine acute T-cell lymphoblastic leukemia cell 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 ral Guanine Nucleotide Exchange Factor small molecule inhibitor stem stem cells synergism technology platform

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.

项目摘要/摘要儿童期急性淋巴细胞白血病（全部）是骨髓的疾病，未成熟细胞并用非特异性化疗治疗。所有这些仍然是癌症死亡的主要原因之一 <20岁的人。 T细胞全部（T-All）的发育起源是诊所的深刻影响。遗传突变指出了异常的RAS信号，在50％的T-all中具有因果作用，但是这种异常是如何信号影响T-ALL中的发育轨迹尚不清楚。同样，这是很大的未知有效抑制异常的RAS信号。在1R01CA187318期间，我们在RAS信号传导中发现了一个新的分子范式。致癌Ras RAS激活剂RASGRP1的突变（KRASG12D）和过表达驱动两个非常独特的白血病 RAS信号。在我们的新rolorig/mx1cre鼠标模型的进度报告中，我们证明了这两个异常的RAS信号都诱导异常的造血，但具有相反的干细胞特征。大写在我们的定量，多个流式细胞仪平台上，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，我们进行了四个具有PI3K抑制剂的广泛合成致死性筛选，以鉴定T-ALL中有效的联合疗法。我们确认使用小分子抑制剂和PI3K抑制剂GDC0941的十种预测组合疗法微管蛋白抑制剂长春新碱可在五种癌症类型中产生广泛的协同作用，在我们的临床前小鼠试验中体内。屏幕预测许多代谢靶标，这仍然是出乎意料的。我们的续签专注于理解异常的RAS-PI3K信号与单元之间的联系代谢的目的是解决T-All的发展起源并测试新组合疗法。我们将在我们的面板中确定RAS-PI3K信号与细胞代谢之间的连接 T-ALL细胞系通过研究屏幕中鉴定出的六个营养转运蛋白（SLC）（AIM 1）。我们将获得对信号，代谢特征和造血组成的机械理解，轨迹，作为白血病RAS-PI3K信号的函数，具有前所未有的分辨率。我们将继续努力在Rolorig/MX1CRE和KRASG12D/MX1CRE鼠标模型上，显示完全相反的表型骨髓干细胞（AIM 2）。最后，我们将获得对发展的全面表征带有单细胞分辨率的小儿T-All的生化和代谢程序。我们将结合这些临床前试验的努力测试PI3Kγ-和δ-抑制作用的体内效率，抑制SLC13A2和 SLC25A44，以及我们的T-All/NGS平台中的组合（AIM 3）。协同目标，代谢见解，新的鼠标模型，患者样品NSG管道和创新的单细胞分辨率技术平台（cytof，scenith，phosho-flow）将使我们能够为分子做出重大贡献了解癌症中T-All和精确医学。