Role of Hedgehog Signaling in JAK2V617F Associated Myelofibrosis

Hedgehog 信号转导在 JAK2V617F 相关骨髓纤维化中的作用

• 批准号: 9916647
• 负责人: Akil Merchant
• 金额: $ 42.5万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9916647
• 关键词: Acute Myelocytic Leukemia; Adult; Affect; Alleles; Animal Model; Autoimmune Diseases; Award; Binding; Blood; Bone Marrow; Cells; Chronic Kidney Failure; Clinical; Complex; Cytometry; Disease; Embryonic Development; Erinaceidae; Event; FRAP1 gene; Fibrosis; Genes; Image; Imaging Techniques; Immune; Immune response; Inflammation; JAK2 gene; Knockout Mice; Ligands; Liver Cirrhosis; MADH2 gene; MAP Kinase Gene; MEKs; Malignant Neoplasms; Marrow; Mediating; Megakaryocytes; Modeling; Molecular; Mus; Mutation; Myelofibrosis; Myeloid Cells; Myeloproliferative disease; NF-kappa B; Non-Malignant; Oncogenic; Paracrine Communication; Pathogenesis; Pathway interactions; Patients; Phenotype; Polycythemia Vera; Primary Myelofibrosis; Production; Proto-Oncogene Proteins c-akt; Pulmonary Fibrosis; Reaction; Recurrence; Regulation; Research; Resolution; Role; SHH gene; Sampling; Signal Transduction; Source; Splenomegaly; Stromal Cells; Therapeutic; Tissues; Transforming Growth Factor beta; Transgenic Mice; Transgenic Organisms; Transplantation; United States; anti-tumor immune response; autocrine; cytokine; disease phenotype; inhibitor/antagonist; injury and repair; mutant; paracrine; promoter; smoothened signaling pathway; targeted treatment; therapeutic target; tumor

Myelofibrosis (MF) is the deadliest of the various myeloproliferative neoplasms (MPN) that affect approximately 150,000 people in the United States. MPN are characterized by abnormal proliferation of one or more of the blood lineages, bone marrow fibrosis, splenomegaly, and progression to acute myeloid leukemia. The identification of recurrent mutations in these patients, the most common being the JAK2V617F mutation found in 95% of polycythemia vera (PV) and 65% of primary MF (PMF) patients, suggests a common molecular pathogenesis centered on aberrant JAK/STAT signaling; however, JAK inhibitors alone offer only modest clinical benefit, without cure. Combinations of JAK inhibitors with other inhibitors of oncogenic pathways such as AKT, MEK/ERK, mTOR, or Hedgehog (Hh), show greater efficacy then JAK inhibitors alone, suggesting that a complex signaling network drives MPN. During my K08 award period, we have demonstrated that: 1) JAK2V617F transgenic mice show increased sonicHh (Shh) ligand expression and activation of Smoothened (Smo)/Hh signaling 2) Treatment with the Hh/Smo inhibitor, PF-04449913 (PF-913, glasdegib), reduces the splenomegaly, cytokine production, marrow fibrosis and JAK2V617F allele burden 3) JAK2V617F transgenic tissues show increased MAPK and NFKB signaling as well as increased TGF-B levels, which decrease with Smo inhibition 4) JAK2V617F mutant cells cause Hh ligand dependent activation of Hh target genes and TGF-B/SMAD2 signaling in stromal cells. Therefore, we hypothesize that Hedgehog signaling is essential for JAK2V617F driven diseases and represents and important therapeutic target. The aims of this study are to determine how JAK2V617F leads to Hh pathway activation and delineate the autocrine vs paracrine effects of abnormal Hh signaling in the bone marrow microenvironment. We will focus on understanding how TGF-B mediates fibrosis and alters the host immune response to MPN. This study will make use of a breakthrough imaging technique known as Imaging Mass Cytometry, to characterize with 40+ parameters the paracrine signaling events and immune response that drives the fibrosis reaction. Understanding the interaction between mutant JAK signaling and hedgehog signaling has implications beyond MPN, as abnormal JAK/STAT and Hedgehog signaling have been implicated in numerous cancers as well as in non-cancerous conditions such as autoimmune diseases, graft verses host disease, inflammation and liver cirrhosis.

骨髓纤维化 (MF) 是影响骨髓增生性肿瘤 (MPN) 的最致命的一种 美国约有 150,000 人。 MPN 的特点是异常 一种或多种血统的增殖、骨髓纤维化、脾肿大和 进展为急性髓系白血病。这些中反复发生的突变的鉴定 患者中，最常见的是 95% 的真性红细胞增多症中发现的 JAK2V617F 突变 (PV) 和 65% 的原发性 MF (PMF) 患者，表明存在共同的分子发病机制 以异常的 JAK/STAT 信号传导为中心；然而，JAK 抑制剂单独提供的效果有限 临床获益，无需根治。 JAK抑制剂与其他致癌抑制剂的组合 AKT、MEK/ERK、mTOR 或 Hedgehog (Hh) 等途径比 JAK 表现出更高的功效 单独的抑制剂，表明 MPN 是由一个复杂的信号网络驱动的。在我的K08期间 颁奖期间，我们已经证明：1) JAK2V617F 转基因小鼠表现出增加 sonicHh (Shh) 配体表达和 Smoothened (Smo)/Hh 信号传导的激活 2) 使用 Hh/Smo 抑制剂 PF-04449913（PF-913，glasdegib）治疗可减少 脾肿大、细胞因子产生、骨髓纤维化和 JAK2V617F 等位基因负担 3) JAK2V617F 转基因组织显示出 MAPK 和 NFKB 信号传导增强以及 TGF-B 水平，随着 Smo 抑制而降低 4) JAK2V617F 突变细胞导致 Hh 配体 基质细胞中 Hh 靶基因和 TGF-B/SMAD2 信号传导的依赖性激活。 因此，我们假设 Hedgehog 信号对于 JAK2V617F 驱动至关重要 疾病并代表重要的治疗靶点。本研究的目的是 确定 JAK2V617F 如何导致 Hh 通路激活并描绘自分泌与 骨髓微环境中异常 Hh 信号传导的旁分泌效应。我们将重点 了解 TGF-B 如何介导纤维化并改变宿主对 MPN 的免疫反应。 这项研究将利用一种称为成像质量的突破性成像技术 细胞计数，用 40 多个参数表征旁分泌信号事件和免疫 驱动纤维化反应的反应。了解突变型 JAK 之间的相互作用 信号传导和刺猬信号传导的影响超出了 MPN，因为异常的 JAK/STAT 和 Hedgehog 信号传导与多种癌症以及非癌性病变有关 自身免疫性疾病、移植物抗宿主病、炎症和肝脏等疾病 肝硬化。