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.

急性髓系白血病 (AML) 是成人中最致命的血液恶性肿瘤，主要是由于高发病率 化疗后复发。残留疾病，免疫表型、形态学上的持续存在， 或一线化疗后基因定义的白血病细胞，是 AML 的不良预后因素。 在小鼠模型中，残留的 AML 细胞已被证明定位于骨内膜骨髓区域，其中 他们可能利用专门支持长期静止造血干细胞的微解剖学生态位 进入休眠状态并获得对针对分裂细胞的化疗的抵抗力。然而，这些利基市场 人类骨髓中的定义不明确，目前尚不清楚人类 AML 细胞是否真的与 典型的干细胞生态位作为逃避化疗的机制。我们的中心假设是 AML 细胞 通过过度表达干细胞生态位保留来与骨髓内保护性生态位中的细胞相互作用 受体，这使它们能够获得化学抗性。在目标 1 中，我们将采用表位联合索引 （CODEX），一种最近开发的方法，可在单细胞水平上可视化数十种抗体标记物 固定人体组织，解剖人体骨髓生态位的确切组成和空间组织。 我们将首次能够确定化疗耐药细胞在临床骨髓中的定位 AML 患者的活检。我们将评估非分裂残留 AML 细胞是否局部化 优先进入骨髓中的骨内膜干细胞生态位以响应化疗。在目标 2 中，我们将 确定干细胞生态位保留受体是否在残留的 AML 细胞上过度表达并促进 化学耐药性。为了实现这一目标，在目标 2A 中，我们将首先对 AML 骨进行单细胞 RNA 测序 诊断时的骨髓抽吸样本和残留疾病测定的时间，以及健康者 对照，提供 AML 细胞、AML 相关微环境细胞及其表达谱 健康的同行。我们将确定介导生态位保留的受体是否过度表达。在 目标 2B，我们将使用 CRISPR-Cas9 评估它们对于化疗耐药性是否具有功能重要性 介导敲除过度表达的受体-配体对，这些受体-配体对介导 AML-基质相互作用 将用细胞毒性化疗治疗的风暴共培养系统。如果 AML 细胞确实定位于这些 骨内膜微环境并通过干细胞中受体-配体介导的相互作用获得化疗耐药性 利基，这些受体/配体对将代表化学增敏的有吸引力的目标，这可能有助于 降低 AML 复发风险。总之，该项目准备评估白血病细胞是否劫持健康细胞 干细胞生态位来逃避标准护理化疗，这项工作有可能揭示新的 更好地治疗这种致命疾病的治疗机会。