The function of MS4A3 in normal and malignant hematopoiesis

MS4A3在正常和恶性造血中的功能

• 批准号: 10593588
• 负责人: Michael W. Deininger
• 金额: $ 46.66万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10593588
• 关键词: ABL1 gene; Acute; Acute leukemia; Affect; Aggressive-Phase Myeloid Leukemia; Apoptosis; Attenuated; Blast Phase; Blood; Bone Marrow Transplantation; CD34 gene; Cell Cycle; Cell Differentiation Induction; Cells; Chronic Myeloid Leukemia; Chronic-Phase Myeloid Leukemia; Clinical; Constitution; Constitutional; Cytokine Receptors; Cytokine Signaling; Data; Dependence; Endocytosis; Family; Fluorouracil; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoiesis; Hematopoietic; Hematopoietic System; Human; IL3 Gene; Image; In Vitro; Knock-out; Leukemic Cell; Malignant - descriptor; Maps; Mass Spectrum Analysis; Mediating; Membrane; Messenger RNA; Mouse Strains; Mus; Myelogenous; Myeloid Cells; Normal Cell; Outcome; Pathway interactions; Patients; Phase; Phenotype; Philadelphia Chromosome; Phosphotransferases; Physiological; Prevalence; Printing; Protein Array; Protein Tyrosine Kinase; Proto-Oncogene Proteins c-abl; Publishing; Recurrence; Relapse; Reporting; Residual state; Resistance; Role; Signal Pathway; Signal Transduction; Signaling Protein; Stress; Testing; Therapeutic Agents; Tumor Suppressor Proteins; Tyrosine Kinase Inhibitor; Undifferentiated; Work; Xenograft procedure; biological adaptation to stress; chronic myeloid leukemia cell; cofactor; cytokine; drug sensitivity; improved; in vivo; inhibitor; inhibitor therapy; knock-down; leukemia; leukemic stem cell; leukemogenesis; live cell imaging; member; nanoparticle; novel; overexpression; prevent; receptor; response; single-cell RNA sequencing; stem; stem cells; therapy resistant; transcriptome

Chronic myeloid leukemia (CML) is caused by BCR-ABL1, a constitutively active tyrosine kinase generated from the Philadelphia chromosome. In the chronic phase of CML (CP-CML), myeloid cells are expanded, but maintain terminal differentiation. Most CP-CML patients achieve durable responses to BCR-ABL1 tyrosine kinase inhibitors (TKIs), and their extended survival is reflected by a steep rise in CML prevalence. Unfortunately, TKIs fail to eliminate quiescent CML stem cells (LSCs) whose survival is independent of BCR-ABL1, necessitating lifelong TKI therapy to prevent CML recurrence. In 5-10% of patients, a differentiation block converts CP-CML into blast phase CML (BP-CML), an aggressive acute leukemia that is often BCR-ABL1-independent and TKI resistant. Our overarching hypothesis is that blocked differentiation is central to the BCR-ABL1 independence that characterizes the extremes of the clinical CML spectrum: Persistence of residual leukemia despite TKI therapy and TKI-resistant BP-CML. We have discovered that expression of MS4A3, a member of the MS4A (membrane-spanning four A) family of signaling proteins, is profoundly reduced in quiescent, TKI resistant and BP CML cells, and that low MS4A3 correlates with shorter survival. MS4A3 knockdown (KD) in CML CD34+ cells inhibits myeloid differentiation, and promotes TKI resistance, while ectopic MS4A3 expression has opposite effects (Zhao et al. Blood. 2021;Epub ahead of print. PMID: 34780648). Our preliminary data suggest that MS4A3 promotes IL-3 and GM-CSF signaling in CML CD34+ cells by promoting endocytosis of their cognate β common chain (βc) receptors. We hypothesize that MS4A3 promotes myeloid differentiation by enhancing response to βc cytokines in leukemic stem and progenitor cells (LSPCs). TKI resistant CML cells downregulate MS4A3 to blunt response to differentiation-inducing cytokines, thereby maintaining a primitive, therapy-resistant state. Re- establishment of MS4A3 expression will enforce differentiation and enhance drug sensitivity. In Aim 1, we will delineate how MS4A3 regulates endocytosis and signaling of βc cytokine receptors. We will track endocytosis by high-throughput immunofluorescent and confocal live cell imaging, identify MS4A3 cofactors by mass spectrometry, and MS4A3-regulated signaling pathways by reverse phase protein array. In Aim 2, we will determine the function of MS4A3 in normal hematopoiesis. We will generate mouse strains with hematopoietic- specific conditional Ms4a3 knockout or inducible overexpression and characterize their hematopoietic system at steady state and under stress. In Aim 3, we will delineate the role of MS4A3 in CML hematopoiesis and as a therapeutic agent in CML. We will test whether modulating MS4A3 expression in LSPCs affects leukemogenesis and TKI response, and whether MS4A3-loaded nanoparticles attenuate BP-CML in xenografts. If successful, we will establish MS4A3 as a novel master regulator of βc cytokine signaling that governs signal strength by modulating endocytosis. BP-CML remains mostly incurable, and most CP-CML patients require lifelong TKI therapy. Our work may provide proof of concept for using forced MS4A3 expression to overcome TKI resistance.

慢性髓样白血病（CML）是由BCR-ABL1引起的，BCR-ABL1是由由组成症活性的酪氨酸激酶产生的 费城染色体。在CML（CP-CML）的慢性阶段，髓样细胞扩展，但保持 终端分化。大多数CP-CML患者对BCR-ABL1酪氨酸激酶的持久反应 抑制剂（TKI）及其延长的存活率反映了CML患病率的钢升高。不幸的是，TKIS 无法消除生存与BCR-ABL1独立的静态CML干细胞（LSC），必要 终身TKI疗法可防止CML复发。在5-10％的患者中，分化块转化了CP-CML 进入BLAST相CML（BP-CML），这是一种侵略性急性白血病，通常是BCR-ABL1独立的，TKI 抵抗的。我们的总体假设是，阻塞的分化对于BCR-ABL1独立性至关重要 这是临床CML光谱的极端的特征：残留白血病任务的持久性TKI 治疗和抗TKI抗BP-CML。我们发现MS4A的成员MS4A3的表达 （跨膜四个a）信号蛋白家族在静止，抗TKI和 BP CML细胞，低MS4A3与较短的存活相关。 CML CD34+细胞中的MS4A3敲低（KD） 抑制髓样分化并促进TKI耐药性，而Ecopic MS4A3表达相反 效果（Zhao等人血液。2021; Epub在印刷前。PMID：34780648）。我们的初步数据表明MS4A3 通过促进其同源β常见的内吞作用，促进CML CD34+细胞中的IL-3和GM-CSF信号传导 链（βC）受体。我们假设MS4A3通过增强对βC的反应来促进髓样分化 白血病和祖细胞（LSPC）中的细胞因子。 TKI抗性CML细胞将MS4A3下调至蓝色 关于- MS4A3表达的建立将实施分化并增强药物敏感性。在AIM 1中，我们将 描述MS4A3如何调节βC细胞因子受体的内吞和信号传导。我们将追踪内吞作用 通过高通量免疫荧光和共聚焦活细胞成像，通过质量识别MS4A3辅助因子 通过反相蛋白阵列通过光谱法和MS4A3调节的信号通路。在AIM 2中，我们将 确定MS4A3在正常造血剂中的功能。我们将产生造血的小鼠菌株 特定的条件MS4A3敲除或诱导的过表达，并表征其造血系统 稳态和压力下。在AIM 3中，我们将描述MS4A3在CML造血中的作用，作为一个 CML中的治疗剂。我们将测试在LSPC中调节MS4A3表达是否影响白血病 和TKI响应，以及MS4A3负载的纳米颗粒是否减弱了异种移植物中的BP-CML。如果成功，我们 将建立MS4A3作为βC细胞因子信号的新型主调节剂，该信号通过 调节内吞作用。 BP-CML仍然无法治愈，大多数CP-CML患者都需要终身TKI 治疗。我们的工作可能会提供概念证明，用于使用强制的MS4A3表达来克服TKI抗性。