Investigation of the impact of pegylated Interferon on clonal trajectory and inflammatory cytokine production in MPN patients

聚乙二醇干扰素对 MPN 患者克隆轨迹和炎性细胞因子产生的影响的研究

• 批准号: 10357190
• 负责人: Andrew Jeffrey Dunbar
• 金额: $ 24.82万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10357190
• 关键词: Acute leukemia; Alleles; Attenuated; Biological Assay; Biology; Bone Marrow; Cell Compartmentation; Cell Cycle; Cell division; Cells; Clinical; Clonal Hematopoietic Stem Cell; DNA sequencing; Data; Disease; Erythroid; Erythroid Cells; Event; Gene Expression Profile; Gene Expression Profiling; Genetic Transcription; Hematology; Hematopoiesis; Hematopoietic stem cells; Hemorrhage; Hemorrhagic Thrombocythemia; Heterogeneity; Human; Individual; Inflammation; Inflammatory; Interferons; Investigation; JAK2 gene; Link; Megakaryocytes; Molecular; Mononuclear; Mutation; Myelofibrosis; Myeloid Cells; Myeloproliferative disease; Output; Pathologic; Pathway interactions; Patients; Phase; Play; Polycythemia Vera; Population; Production; Reporting; Risk; Role; Sampling; Serum; Signal Pathway; Signal Transduction; Symptoms; Work; cytokine; detection limit; disorder control; exhaustion; experimental study; fitness; hydroxyurea; insight; interferon therapy; mutant; neutrophil; patient subsets; peripheral blood; phase 2 study; phase 3 study; phase III trial; predict clinical outcome; progenitor; responders and non-responders; response; response biomarker; single cell technology; stem; thrombotic; transcriptomics; treatment response

PROJECT SUMMARY The myeloproliferative neoplasms (MPNs) which include Polycythemia Vera (PV), Essential Thrombocythemia (ET) and Myelofibrosis (MF) are clonal hematopoietic stem cell disorders molecularly characterized by aberrant activation of the JAK-STAT signaling pathway. The JAK-STAT pathway is most often activated by mutations in JAK2, CALR, or MPL. Biologically, theses disease are characterized by proliferation of mature myeloid and erythroid cells, and overproduction of inflammatory cytokines. Clinically, these diseases result in increased risk of thrombotic and hemorrhagic events, progressive bone marrow fibrosis, a high degree of symptom burden, and ultimately transformation to acute leukemia. Cytoreductive therapy, including hydroxyurea (HU) and pegylated Interferon (pegINF) are commonly used agents used to control disease manifestations in patients with ET and PV. Notably, pegINF has demonstrated the ability to procure clinical response (including hematologic and pathologic response in patients with ET and PV). Further, peripheral blood DNA sequencing studies of mature neutrophils and mononuclear cells have demonstrated that pegINF can results in decreases of JAK2V617F allele burden, in some cases to levels below the limits of detection of employed assays. However, the mechanisms by which INF exerts its effects in MPN, including its effects on MPN hematopoietic stem and progenitor cells (HSPCs), remains to be resolved. The observation that pegINF therapy can normalize peripheral blood counts, decrease mutant JAK2 allele burden in the peripheral blood, and alter cytokine expression in the serum of MPN patients suggests the possibility that pegINF may work by depleting JAK2 mutant HSPCs, alter the fitness of JAK2 mutant HSCs, or alter the transcriptional profile of HSPCs to reduce megakaryocyte/erythroid lineage bias. However, no comprehensive analysis of the impact of pegINF therapy on mutant and wildtype HPSCs in MPN patients has been reported. We hypothesize that pegINF therapy results in hematologic and molecular responses by reducing the clonal output of JAK2mut HSPCs and attenuates inflammatory cytokine production by reducing the number of mature and immature myeloid and erythroid cells derived from JAKmut HSPCs as well as the cytokine production per cell. Using primary patient samples for the recently reported phase II study of pegINF in ET and PV patients (MPD-RC 111 study) and the Phase III trial of hydroxyurea versus INF in untreated PV and ET patients (MPD- RC 112 study) we will undertake single-cell approaches to determine the impact of pegINF on the lineage trajectory of JAK2 mutant HSPCs, the transcriptional output of HSPCs, and on cytokine production by these populations. Our experiments will reveal insights into how pegINF alters hematopoiesis and inflammation in the bone marrow of MPN patients, and identify predictors of disease response.

项目概要 骨髓增生性肿瘤 (MPN)，包括真性红细胞增多症 (PV)、原发性 血小板增多症（ET）和骨髓纤维化（MF）是分子克隆性造血干细胞疾病 其特征是 JAK-STAT 信号通路的异常激活。 JAK-STAT 途径最常见 由 JAK2、CALR 或 MPL 突变激活。从生物学角度来看，这些疾病的特点是增殖 成熟的骨髓细胞和红细胞的数量，以及炎症细胞因子的过量产生。临床上，这些疾病 导致血栓和出血事件、进行性骨髓纤维化、高度 症状负担，最终转变为急性白血病。细胞减灭疗法，包括 羟基脲 (HU) 和聚乙二醇化干扰素 (pegINF) 是控制疾病的常用药物 ET 和 PV 患者的表现。值得注意的是，pegINF 已证明有能力获得临床 反应（包括 ET 和 PV 患者的血液学和病理学反应）。此外，外围 成熟中性粒细胞和单核细胞的血液 DNA 测序研究表明，pegINF 可导致 JAK2V617F 等位基因负担减少，在某些情况下降低至低于检测限的水平 采用的分析方法。然而，INF 在 MPN 中发挥作用的机制，包括其对 MPN 造血干细胞和祖细胞 (HSPC) 仍有待解决。 pegINF治疗可使外周血计数正常化、减少突变型JAK2的观察结果 外周血中的等位基因负荷以及 MPN 患者血清中细胞因子表达的改变表明 pegINF 可能通过消耗 JAK2 突变型 HSPC 发挥作用，改变 JAK2 突变型 HSC 的适应性，或 改变 HSPC 的转录谱以减少巨核细胞/红细胞谱系偏差。然而，没有 综合分析pegINF治疗对MPN患者突变型和野生型HPSC的影响 被举报。我们假设 pegINF 治疗通过以下方式导致血液学和分子反应： 减少 JAK2mut HSPC 的克隆输出并减弱炎症细胞因子的产生 减少源自 JAKmut HSPC 的成熟和未成熟骨髓细胞和红细胞的数量 以及每个细胞的细胞因子产量。 使用主要患者样本进行最近报道的 pegINF 在 ET 和 PV 患者中的 II 期研究 （MPD-RC 111 研究）以及在未经治疗的 PV 和 ET 患者中羟基脲与 INF 的 III 期试验（MPD- RC 112 研究）我们将采用单细胞方法来确定 pegINF 对谱系的影响 JAK2 突变体 HSPC 的轨迹、HSPC 的转录输出以及这些细胞产生的细胞因子 人口。我们的实验将揭示 pegINF 如何改变造血和炎症 MPN 患者的骨髓，并确定疾病反应的预测因子。