Trajectory and Architecture of Microenvironment-Mediated Resistance in AML

AML 中微环境介导的耐药性的轨迹和结构

• 批准号: 10517761
• 负责人: Anupriya Agarwal
• 金额: $ 36.96万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10517761
• 关键词: ATAC-seq; Acute Myelocytic Leukemia; Address; Aftercare; Architecture; BCL2 gene; Biology; Bone Marrow; CRISPR screen; Cell Line; Cell Survival; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Combined Modality Therapy; Communication; Complex; DNA sequencing; Data; Data Set; Development; Drug Combinations; Drug Sensitization; Drug resistance; Engineering; Environment; Epigenetic Process; Event; Evolution; Exposure to; FGF2 gene; FLT3 gene; FLT3 inhibitor; Flow Cytometry; Funding; Gene Expression; Genetic Heterogeneity; Goals; Growth; Growth Factor; Heterogeneity; Immune; Immune Evasion; Immune Targeting; Immune signaling; Immunofluorescence Immunologic; Immunologic Factors; Inflammatory; Interleukin-1; JAK2 gene; Leukemic Cell; MEKs; Malignant - descriptor; Maps; Mediating; Medical Genetics; Methylation; Modeling; Molecular; Multiomic Data; Mutation; Natural Killer Cells; Organoids; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Play; Process; Proteomics; Recurrent disease; Refractory; Refractory Disease; Relapse; Residual state; Resistance; Role; Route; Sampling; Screening Result; Signal Pathway; Signal Transduction; Stress; Stromal Cells; T-Lymphocyte; Testing; Therapeutic; Therapeutic Intervention; acquired drug resistance; acute myeloid leukemia cell; base; cancer cell; cytokine; data integration; deep sequencing; disorder control; drug development; drug sensitivity; genome-wide; improved; inhibitor; leukemia; monocyte; neoplastic cell; network models; new combination therapies; novel; novel therapeutic intervention; predictive modeling; progenitor; resistance mechanism; response; screening; single-cell RNA sequencing; small molecule inhibitor; targeted treatment; therapeutic target; therapy resistant; transcriptome sequencing; transcriptomics; translational impact; treatment response; tumor; tumor microenvironment

PROJECT SUMMARY/ABSTRACT: Project 2 Outcomes for acute myeloid leukemia (AML) patients have improved little over four decades. This is largely due to the development of acquired drug resistance and refractory disease. Thus, there is an urgent need for new strategies to target residual AML cells or refractory clones before they trigger disease relapse. The tumor microenvironment is a key factor in the development of malignant progression and therapy resistance. The bi- directional communication between cancer cells and microenvironmental cells is much more complex than initially perceived. Thus, one strategy to reduce relapse is to target the signaling mediated by growth factors secreted by stromal or immune cells within the bone marrow microenvironment that play a critical role in promoting leukemia cell survival, development of drug resistance, and immune evasion. Therefore, the long- term goal of this project is to deconvolute the comprehensive network of events and cellular heterogeneity that contribute to acquired resistance in a context dependent manner to identify new therapeutic approaches. As part of our prior U54 DRSN funding, omics-based analysis of 350 primary AML patient samples found that multiple secreted cytokines and immune factors may contribute to drug resistance. Therefore, we hypothesize that reprogramming of the bone marrow niche modulates drug response and drives acquired resistance in AML. Comprehensive understanding of these mechanisms will lead to the identification of new combination therapies. We will focus on 5 essential therapeutic targets in AML: BCL2, FLT3, JAK2, MEK, and epigenetic pathways and will address this hypothesis following three well integrated aims. In Aim 1, we will map microenvironmental signatures and tumor cross-talk mechanisms against drug response features of AML by performing single cell gene expression and epigenetic analyses in the context of bone marrow niche. Tumor microenvironment cross- talk will be further explored in Aim 2 focusing on monocytes and stromal cells and in the context of NK and T cells in Aim 3 by performing CRISPR/Cas screening with co-culture platforms. Identified targets/pathways will be validated to fully define microenvironmental mechanisms of acquired drug resistance. These results will build predictive models of signaling crosstalk in response to therapeutic stress and identify combination therapies to overcome acquired resistance, particularly in the context of the microenvironment. We will test the effect of these inhibitors on cell viability, cellular heterogeneity, differentiation, and target inhibition using multi-parametric flow cytometry and immunofluorescence analysis. We will integrate these findings with Project 1 and prioritized targets will be tested for their translational impact in Project 3. Overall, elucidating the influence of microenvironment-driven signaling on drug response and survival of AML cells will help identify novel tractable targets for combination therapies to overcome acquired drug resistance in AML.

项目总结/摘要：项目2 急性髓性白血病（AML）患者的预后在过去40年中几乎没有改善。这主要是由于 获得性耐药性和难治性疾病的发展。因此，迫切需要新的 针对残留的AML细胞或难治性克隆，在它们触发疾病复发之前，肿瘤 微环境是恶性进展和治疗抗性发展的关键因素。双- 癌细胞和微环境细胞之间的定向通讯比 最初被感知。因此，减少复发的一个策略是靶向由生长因子介导的信号传导 由骨髓微环境中的基质细胞或免疫细胞分泌，在 促进白血病细胞存活、产生耐药性和免疫逃避。因此，长期以来- 该项目的长期目标是去卷积事件和细胞异质性的综合网络， 以背景依赖的方式促进获得性耐药性，以鉴定新的治疗方法。一部分 在我们之前的U54 DRSN资金中，对350例原发性AML患者样本进行的基于omics的分析发现， 分泌的细胞因子和免疫因子可能导致耐药性。因此，我们假设 在AML中，骨髓小生境的重编程调节药物反应并驱动获得性耐药性。 对这些机制的全面理解将导致新的联合治疗的确定。 我们将专注于AML的5个基本治疗靶点：BCL 2，FLT3，JAK 2，MEK和表观遗传途径， 将在三个综合目标之后讨论这一假设。在目标1中，我们将绘制微环境地图， 针对AML的药物反应特征的信号和肿瘤串扰机制， 基因表达和表观遗传学分析的背景下，骨髓生态位。肿瘤微环境交叉- 我将在目标2中进一步探讨单核细胞和基质细胞，以及NK和T细胞的背景下， 通过用共培养平台进行CRISPR/Cas筛选，在Aim 3中培养细胞。确定的目标/途径将 以充分确定获得性耐药性的微环境机制。这些结果将建立 响应于治疗应激的信号传导串扰的预测模型，并鉴定联合疗法， 克服获得性抗性，特别是在微环境中。我们将测试这些的效果 使用多参数流对细胞活力、细胞异质性、分化和靶抑制的抑制剂 流式细胞术和免疫荧光分析。我们将把这些发现与项目1结合起来， 将在项目3中测试目标的转化影响。总的来说，阐明 微环境驱动的信号对药物反应和AML细胞的存活将有助于确定新的易处理的 联合治疗的靶点，以克服AML的获得性耐药性。