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.

慢性粒细胞白血病（CML）是由BCR-ABL 1引起的，BCR-ABL 1是一种组成型活性酪氨酸激酶，由 费城染色体在慢性期CML（CP-CML），髓系细胞扩增，但维持在正常水平。 终末分化大多数CP-CML患者对BCR-ABL 1酪氨酸激酶实现持久应答 抑制剂（TKI），其延长的生存期反映在CML患病率的急剧上升。不幸的是， 不能消除存活不依赖于BCR-ABL 1的静止期CML干细胞（LSC）， 终身TKI治疗以预防CML复发。在5-10%的患者中，分化阻滞使CP-CML转化为 急变期CML（BP-CML），一种侵袭性急性白血病，通常是BCR-ABL 1非依赖性的， 抵抗我们的总体假设是，阻断分化是BCR-ABL 1独立性的核心。 临床CML谱的极端特征：尽管TKI，残留白血病持续存在 治疗和TKI耐药BP-CML。我们已经发现，MS 4A基因的成员MS 4A 3的表达， （跨膜四A）家族的信号蛋白，在静止期，TKI抗性和 BP CML细胞，低MS 4A 3与较短的生存期相关。CML CD 34+细胞中的MS 4A 3敲低（KD） 抑制骨髓分化，促进TKI耐药，而异位MS 4A 3表达则相反。 效果（Zhao等人，Blood. 2021年;Epub先于印刷。PMID：34780648）。我们的初步数据表明，MS 4A 3 通过促进其同源β共同体的内吞作用促进CML CD 34+细胞中的IL-3和GM-CSF信号传导 链（βc）受体。我们假设MS 4A 3通过增强对βc的反应促进髓系分化， 白血病干细胞和祖细胞（LSPC）中的细胞因子。TKI耐药CML细胞下调MS 4A 3以钝化 细胞对诱导分化的细胞因子的应答，从而维持原始的治疗抗性状态。再 MS 4A 3表达的建立将促进分化并增强药物敏感性。在目标1中，我们 描述MS 4A 3如何调节βc细胞因子受体的内吞作用和信号传导。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，按质量鉴定MS 4A 3辅因子 质谱和MS 4A 3调节的信号传导途径通过反相蛋白阵列。在目标2中，我们将 确定MS 4A 3在正常造血中的功能。我们会培育出具有造血功能的小鼠- 特异性条件Ms 4a 3敲除或诱导型过表达并表征其造血系统， 稳定状态和压力下。在目的3中，我们将描述MS 4A 3在CML造血中的作用，并作为一种免疫调节剂。 CML的治疗药物。我们将测试调节MS 4A 3在LSPC中的表达是否影响白血病的发生 和TKI反应，以及加载MS 4A 3的纳米颗粒是否减弱异种移植物中的BP-CML。如果成功，我们 将确立MS 4A 3作为βc细胞因子信号传导的新型主调节因子，其通过以下方式控制信号强度： 调节内吞作用。BP-CML仍然大多无法治愈，大多数CP-CML患者需要终身TKI 疗法我们的工作可能为使用强制MS 4A 3表达来克服TKI抗性提供概念证明。