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

项目总结/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增的疾病， 细胞并接受非特异性化疗。ALL仍然是癌症死亡的主要原因之一， 20岁以下的人。T细胞急性淋巴细胞白血病（T-ALL）的发育起源对临床有着深远的影响。 基因突变指出异常Ras信号在50%的T-ALL中起因果作用，但这种异常Ras信号如何在T-ALL中发挥作用？ 信号影响T-ALL的发育轨迹尚不清楚。同样，它是由和大部分未知如何 有效抑制异常Ras信号。 在1 R 01 CA 187318期间，我们发现了Ras信号传导中的一种新的分子模式：致癌Ras 突变（KRasG 12 D）和Ras激活剂RasGRP 1的过表达驱动两种非常独特的白血病细胞。 Ras信号。在我们新的RoLoRiG/Mx 1CRE小鼠模型的进展报告中，我们表明这两个 异常Ras信号均诱导异常造血，但具有相反的干细胞特征。资本化 在我们的定量、多重流式细胞仪平台上，我们确定Ras和RasGRP 1经常在 连接到T-ALL中的PI 3 K（磷酸肌醇3-激酶）-AKT/mTOR信号传导。接下来，我们表演了四个 使用PI 3 K抑制剂进行广泛的合成致死性筛选，以确定T-ALL的有效联合治疗。我们 证实了10种预测的小分子抑制剂和PI 3 K抑制剂GDC 0941联合治疗 与微管蛋白抑制剂阿曲斯汀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， vivo.这些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-PI 3 K信号与细胞凋亡之间的联系。 目的是解决T-ALL的发育起源和测试新的组合 治疗我们将在我们的10人小组中确定Ras-PI 3 K信号与细胞代谢之间的联系 T-ALL细胞系通过研究我们筛选中鉴定的六种营养转运蛋白（SLC）（目的1）。我们将 获得对信号、代谢特征和造血组成的机制性理解， 轨迹，作为白血病Ras-PI 3 K信号的函数，具有前所未有的分辨率。我们将继续努力 在RoLoRiG/Mx 1CRE和KRasG 12 D/Mx 1CRE小鼠模型中， 骨髓干细胞（Aim 2）。最后，我们将获得一个全面的表征发展， 单细胞分辨率的儿科T-ALL的生化和代谢程序。我们将联合收割机 通过临床前试验测试PI 3 K γ-和δ-抑制、SLC 13 A2抑制和 SLC 25 A44和我们的T-ALL/NGS平台中的组合（Aim 3）。协同的目标，代谢的见解， 新的小鼠模型、患者样本-NSG管道和创新的单细胞分辨率技术平台 （CyTOF，SCENITH，磷酸流）将使我们能够对分子生物学做出重大贡献。 了解T-ALL和癌症精准医疗。

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.