Investigating the role of the JAK/STAT3 Pathway in Clonal Proliferation and Immune Dysfunction in Myelodysplastic Syndrome

研究 JAK/STAT3 通路在骨髓增生异常综合征克隆增殖和免疫功能障碍中的作用

基本信息

批准号：
10223409
负责人：
PAUL B FERRELL
金额：
$ 16.27万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2023-07-31
项目状态：
已结题

项目摘要

Project Summary Myelodysplastic syndrome (MDS) is a heterogeneous group of pre-leukemic bone marrow failure disorders with a yearly incidence of approximately 13,000 in the U.S. Approximately one third of MDS patients will go on to develop acute myeloid leukemia (AML) with a dismal survival of 5-10%. Standard of care therapy only extends life by a median of 9 months in those requiring treatment and there is no cure for the disease outside of stem cell transplant. As the population ages, this disease will only become more common, thus there is an urgent need for more effective treatments in MDS. MDS pathogenesis is marked by both clonal myeloid proliferation and bone marrow immune dysfunction. In MDS bone marrow, a suppressive immune microenvironment develops in response to initial inflammation and contributes to the proliferation of developing blasts, which are free to grow unchecked by the immune system. In fact, myeloid-derived suppressor cells (MDSCs), myeloid cells which are suppressive to T cell function, have been implicated in disease pathogenesis and progression. The JAK/STAT3 pathway is critical in myeloid differentiation and is aberrantly activated in both myeloblasts and MDSCs, both of which increase as MDS progresses. In order to better understand JAK/STAT3 signaling in MDS, we will use single cell and functional genomic assays to profile STAT3 in primary patient samples, with particular focus on myeloblasts and MDSCs – two separate but central cells types in leukemic progression of MDS. Given our experience studying single cell phenotype and intracellular signaling in AML, we will adapt our methods to study MDS, with particular focus on p-STAT3. An advantage of this approach is the ability to measure all cells within a sample simultaneously and create single cell signaling profiles for each sample and in response to inhibition. We propose three specific aims to investigate JAK/STAT3 signaling in MDS. In Aim I, the basal and cytokine-induced responses of STAT3 signaling will be measured in banked MDS samples to understand the role of STAT3 signaling in leukemic progression. Aim II will test functional and signaling responses to various inhibitors of the JAK/STAT3 pathway. Finally, Aim III will determine the impact of standard therapy on JAK/STAT3 signaling and STAT3 targets in resistant MDS. Chromatin immunoprecipitation (ChIP-seq) will be used here in order to profile the gene targets of STAT3 in MDS. These studies will significantly contribute to our basic understanding of MDS and the therapeutic potential of the JAK/STAT3 pathway moving forward in this disease. This proposal is ultimately designed to allow Dr. Ferrell to develop further skills in mass cytometry, phospho- flow, immunology techniques and ChIP and to provide him with the critical mentoring and training necessary to become an independent physician-scientist.

项目摘要骨髓增生综合征（MDS）是一组异质性的骨髓骨髓衰竭障碍疾病在美国，每年约有13,000人的事件将继续发展急性髓样白血病（AML），其生存率为5-10％。护理疗法仅扩展在需要治疗的人中，中位数为9个月，无法治愈茎以外的疾病细胞移植。随着人口的年龄，这种疾病只会变得更加普遍，因此很紧急需要在MDS中进行更有效的治疗。 MDS发病机理由两种克隆髓样增殖标记和骨髓免疫功能障碍。在MDS骨髓中，一种抑制性免疫微环境响应初始炎症而发展，并有助于发展爆炸的扩散，这是免疫系统可以自由生长。实际上，髓样衍生的抑制细胞（MDSC），髓样细胞抑制T细胞功能的，在疾病的发病机理和进展中已隐含。 JAK/STAT3途径在髓样分化中至关重要，并且在肌细胞中都被异常激活 MDSC，两者随着MDS的进展而增加。为了更好地了解MDS中的JAK/STAT3信号，我们将使用单细胞和功能性基因组测定来介绍主要患者样本中的STAT3，特别是专注于骨髓细胞和MDSC - 两种单独但中央细胞类型的MDS进展。给出我们研究AML中研究单细胞表型和细胞内信号传导的经验，我们将使我们的方法适应研究MD，特别关注P-Stat3。这种方法的优点是能够测量所有细胞在样本中简单地在每个样本中创建单个单元信号剖面，并响应于抑制。我们提出了三个特定的目的，以研究MDS中的JAK/STAT3信号。在目标一世中，基本和细胞因子诱导的STAT3信号反应将在银行的MDS样本中测量，以了解 STAT3信号在白血病进展中的作用。 AIM II将测试对各种的功能和信号响应 JAK/STAT3途径的抑制剂。最后，AIM III将确定标准疗法对JAK/STAT3的影响抗性MDS中的信号传导和STAT3靶标。染色质免疫沉淀（CHIP-SEQ）将在这里使用为了概述MDS中STAT3的基因靶标。这些研究将极大地促进我们的基本对MDS的了解以及JAK/STAT3途径在该疾病中前进的治疗潜力。该建议最终旨在允许Ferrell博士发展质量细胞仪，磷酸化的进一步技能。流动，免疫学技术和芯片，并为他提供必要的关键心理和培训成为独立的身体科学家。