Determining the Microenvironmental Contribution to Acute Myeloid Leukemia Chemoresistance

确定微环境对急性髓系白血病化疗耐药性的影响

• 批准号: 10602784
• 负责人: Shovik Bandyopadhyay
• 金额: $ 3.52万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10602784
• 关键词: Ablation; Acute Myelocytic Leukemia; Acute leukemia; Adult; Antibodies; Biological Assay; Biopsy; Bone Marrow; Bone Marrow Aspiration; Bone Marrow Cells; Bone Marrow Neoplasms; Bone marrow biopsy; CRISPR/Cas technology; CXCR4 gene; Cell Communication; Cells; Cellularity; Chemoresistance; Chemosensitization; Clinical; Coculture Techniques; Complement; Cytarabine; Cytotoxic Chemotherapy; Detection; Diagnosis; Diagnostic; Disease; Epitopes; Genetic; Genetic Transcription; Goals; Hematologic Neoplasms; Hematopoiesis; Hematopoietic stem cells; Human; Image; Immunologic Deficiency Syndromes; Immunophenotyping; In Situ; Knock-out; Leukemic Cell; Ligands; Marrow; Measures; Mediating; Mediator; Membrane Proteins; Methods; Microanatomy; Morphology; Mus; Mutation; NPM1 gene; Patient Selection; Patients; Physiology; Population; Prognosis; Prognostic Factor; Proliferation Marker; Proteins; Relapse; Residual Neoplasm; Residual state; Resistance; Resolution; Sampling; Source; Stromal Cell-Derived Factor 1; Stromal Cells; Survival Rate; System; Time; Tissues; Visualization; Work; acute myeloid leukemia cell; c-myc Genes; cell type; chemotherapy; clinical remission; hematopoietic stem cell quiescence; human tissue; improved; indexing; leukemia; mouse model; multiplexed imaging; mutant; neoplastic cell; new therapeutic target; novel therapeutics; overexpression; patient derived xenograft model; receptor; relapse risk; response; self-renewal; single-cell RNA sequencing; standard of care; stem cell niche; targeted treatment; therapy development; transcriptomics; treatment response; tumor

Acute Myeloid Leukemia (AML) is the most fatal hematologic malignancy in adults, largely due to high rates of relapse following chemotherapy. Residual disease, the persistence of immunophenotypically, morphologically, or genetically defined leukemia cells following frontline chemotherapy, is a poor prognostic factor for AML. Residual AML cells have been shown to localize to endosteal bone marrow regions in murine models, where they may utilize microanatomical niches specialized to support long-term quiescent hematopoietic stem cells to become dormant and gain resistance to chemotherapy, which targets dividing cells. However, these niches are ill-defined in human bone marrow, and it remains unclear whether human AML cells actually interact with canonical stem cell niches as a mechanism to evade chemotherapy. Our central hypothesis is that AML cells interact with cells in protective niches within bone marrow by overexpressing stem cell niche retention receptors, which allows them to acquire chemoresistance. In Aim 1, we will employ Co-Indexing by Epitopes (CODEX), a recently developed method to visualize dozens of antibody markers on a single-cell level from fixed human tissue, to dissect the exact composition and spatial organization of human bone marrow niches. For the first time, we will be able to determine where chemoresistant cells localize in clinical bone marrow biopsies from AML patients. We will assess whether or not non-dividing residual AML cells localize preferentially to endosteal stem cell niches in the bone marrow in response to chemotherapy. In Aim 2, we will determine whether stem cell niche retention receptors are overexpressed on residual AML cells and promote chemoresistance. To achieve this, in Aim 2A we will first perform single-cell RNA sequencing of AML bone marrow aspirate samples from diagnosis and time of residual disease determination, as well as healthy controls, providing an expression profile of AML cells, AML-associated microenvironmental cells, and their healthy counterparts. We will determine whether receptors mediating niche retention are overexpressed. In Aim 2B, we will assess whether they are functionally important for chemoresistance using CRISPR-Cas9 mediated knockout of overexpressed receptor-ligand pairs which mediate AML-stromal interaction in an AML- stormal co-culture system that will be treated with cytotoxic chemotherapy. If AML cells indeed localize to these endosteal niches and gain chemoresistance through receptor-ligand mediated interaction in the stem cell niche, these receptor/ligand pairs would represent an attractive target for chemosensitization, which may help reduce AML relapse risk. Together, this project is poised to evaluate whether leukemic cells hijack healthy stem cell niches to evade standard-of-care chemotherapy, and this work has the potential to reveal new therapeutic opportunities to better treat this deadly disease.

急性髓细胞性白血病（AML）是成年人最致命的血液系统恶性肿瘤，这主要是由于高率 化疗后复发。残留疾病，从形态上，形态学上的免疫表型的持久性， 或一线化学疗法后遗传定义的白血病细胞是AML的预后因素较差。 残留的AML细胞已显示在鼠模型中的内骨骨髓区域，其中 他们可以利用专门支持长期静止造血干细胞的微解剖壁ches 变得休眠并获得对靶向分裂细胞的化学疗法的抵抗力。但是，这些利基是 在人体骨髓中定义不明，目前尚不清楚人类AML细胞是否实际相互作用 规范干细胞生态位作为逃避化学疗法的机制。我们的中心假设是AML细胞 通过过表达的干细胞生态率保留骨髓内的保护性小裂细胞中的细胞 受体，这使他们能够获得化学抗性。在AIM 1中，我们将通过表位采用共同指数 （codex），一种最近开发的方法，可在单细胞水平上可视化数十个抗体标记 固定人体组织，以剖析人体骨髓壁ni的确切组成和空间组织。 我们将第一次能够确定化学剂细胞在临床骨髓中的位置 来自AML患者的活检。我们将评估非分明残留AML单元是否本地化 优先响应化学疗法的骨髓中的内骨干细胞壁ni。在AIM 2中，我们将 确定干细胞生态位保留受体是否在残留AML细胞上过表达并促进 化学抗性。为此，在AIM 2A中，我们将首先执行AML骨的单细胞RNA测序 从诊断和确定残留疾病的时间和健康的骨髓吸入样品 对照，提供AML细胞，与AML相关的微环境细胞的表达曲线及其 健康的对手。我们将确定介导利基保留率的受体是否过表达。在 AIM 2B，我们将评估使用CRISPR-CAS9的化学耐药性功能在功能上重要 介导过表达的受体配体的敲除，这些对体配体对介导AML-中介导AML - 基质相互作用 风暴共培养系统将通过细胞毒性化疗进行治疗。如果AML细胞确实本地化为这些 内传壁细分市场并通过干细胞中的受体配体介导的相互作用获得化学耐药性 利基市场，这些受体/配体对将代表化学敏化的有吸引力的目标，这可能有助于 降低AML复发风险。该项目共同评估白血病细胞是否健康 干细胞生态位以逃避护理标准化疗，这项工作有可能揭示新的 治疗机会更好地治疗这种致命疾病。