Uncovering the role for MSI2 network in hematopoietic stem cells

揭示 MSI2 网络在造血干细胞中的作用

基本信息

批准号：
10265561
负责人：
Michael Kharas
金额：
$ 63.27万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10265561
关键词：
Address Adult Anemia Bar Codes Binding Biology Blood Blood Cells Bone Marrow Cell Lineage Cell Therapy Cell division Cells Clinical Colony-Forming Units Assay Communication Data Daughter Defect Development Disease Disease Progression Dysmyelopoietic Syndromes Engraftment Failure Future Gene Expression Genes Genetic Hematological Disease Hematopoiesis Hematopoietic Hematopoietic Neoplasms Hematopoietic System Hematopoietic stem cells Immune system Journals Laboratories Lead Lymphoid Maintenance Malignant Neoplasms Mammalian Cell Maps Mediating Medicine Messenger RNA Methods Molecular Morphology Myelogenous Nature Normal Cell Outcome Pathway interactions Patients Phenotype Process Prognosis Proteomics RNA-Binding Proteins Regulation Role Signal Transduction Therapeutic Therapeutic Intervention Transforming Growth Factor beta Transplantation Work base bone marrow failure syndrome conditional knockout cytopenia hematopoietic hierarchy hematopoietic stem cell differentiation hematopoietic stem cell fate hematopoietic stem cell self-renewal improved outcome in vivo insight leukemia leukemic stem cell leukemogenesis mouse model new technology novel novel therapeutic intervention overexpression progenitor programs self-renewal single-cell RNA sequencing small hairpin RNA stem stem cell engraftment stem cell expansion stem cell self renewal stem cells transcriptome sequencing

Address Adult Anemia Bar Codes Binding Biology Blood Blood Cells Bone Marrow Cell Lineage Cell Therapy Cell division Cells Clinical Colony-Forming Units Assay Communication Data Daughter Defect Development Disease Disease Progression Dysmyelopoietic Syndromes Engraftment Failure Future Gene Expression Genes Genetic Hematological Disease Hematopoiesis Hematopoietic Hematopoietic Neoplasms Hematopoietic System Hematopoietic stem cells Immune system Journals Laboratories Lead Lymphoid Maintenance Malignant Neoplasms Mammalian Cell Maps Mediating Medicine Messenger RNA Methods Molecular Morphology Myelogenous Nature Normal Cell Outcome Pathway interactions Patients Phenotype Process Prognosis Proteomics RNA-Binding Proteins Regulation Role Signal Transduction Therapeutic Therapeutic Intervention Transforming Growth Factor beta Transplantation Work base bone marrow failure syndrome conditional knockout cytopenia hematopoietic hierarchy hematopoietic stem cell differentiation hematopoietic stem cell fate hematopoietic stem cell self-renewal improved outcome in vivo insight leukemia leukemic stem cell leukemogenesis mouse model new technology novel novel therapeutic intervention overexpression progenitor programs self-renewal single-cell RNA sequencing small hairpin RNA stem stem cell engraftment stem cell expansion stem cell self renewal stem cells transcriptome sequencing

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项目摘要

PROJECT SUMMARY/ABSTRACT Hematopoietic stem cells (HSCs) must navigate important cellular fate choices that include a symmetric self-renewal, symmetric commitment or undergo an asymmetric cell division where one of the cells is fated to differentiate. Alterations in this homeostatic program can lead to hematopoietic disorders and malignancies. Myelodysplastic syndromes (MDS) are a heterogeneous set of clonal disorders characterized by ineffective blood cell development. A common pathophysiologic mechanism in MDS is the presence of dysregulated hematopoietic stem and progenitor cells that fail to normally develop into the diverse set of blood cells necessary for normal function. Our laboratory and others identified MUSASHI2 (MSI2) as a central regulator of HSC and hematopoietic progenitor cell self-renewal (Kharas et al. Nature Medicine 2010). Additionally, we identified that Msi2 loss results in a defect in controlling symmetric and asymmetric division, failure to engraft and results in defective maintenance of myeloid lineage biased HSCs in part through control of the TGFβ pathway (Park et al. 2014 Journal of Experimental Medicine). We found that elevated levels of MSI2 expression predicts poor outcome and using a genetic MDS mouse model that overexpresses MSI2 can drive a more aggressive MDS (Taggart et al. 2016 Nature Communications). To determine if MSI2 is part of a regulatory network, we performed proteomics and in vivo shRNA screen for functional regulators of leukemia self-renewal. Based on this screen, we identified SYNCRIP, an RNA binding protein that shares MSI2 targets and is required in leukemia stem cells (Vu et al., 2017 Nature Genetics). Our preliminary data with a conditional knockout for Syncrip indicates that it is also critical for HSC self-renewal. Our proposal will expand our focus from MSI2 to its associated regulatory network and characterize and identify new molecular determinants for HSC and HSPC symmetric self-renewal and asymmetric fate choice. We have adapted new technologies that include a barcoding, single cell RNA-seq and paired daughter HSC assays (FATE-seq). We have also develop a new way to map direct mRNA targets in HSCs called (HYPERTRIBE), (Nguyen et al. Nature Communications 2020).This proposal will identify new regulators of HSPC fate choice which will lead to novel therapeutic strategies to improve outcomes in MDS patients.

项目摘要/摘要造血干细胞（HSC）必须导航重要的细胞命运选择，包括对称自我更新，对称承诺或经历不对称的细胞分裂其中一个细胞被命中分化。该体内稳态程序的改变可能会导致造血疾病和恶性肿瘤。骨髓增生综合征（MDS）是以无效的血细胞发育为特征的异质性克隆疾病。一个 MDS中常见的病理生理机制是造血失调的存在茎和祖细胞通常无法发展成潜水员的血细胞集正常功能所必需的。我们的实验室和其他人确定Musashi2（MSI2）是 HSC和造血祖细胞细胞自我更新的中央调节剂（Kharas等人。 Medicine 2010）。此外，我们确定MSI2损失导致控制缺陷对称和不对称的划分，未植入并导致缺陷维持髓样谱系偏向HSC，部分通过控制TGFβ途径（Park等，2014 实验医学杂志）。我们发现MSI2表达预测水平升高结果不佳并使用过表达MSI2的遗传MDS小鼠模型可以驱动A 更具侵略性的MD（Taggart等人，2016年自然通信）。确定MSI2是否为一部分监管网络，我们进行了蛋白质组学和体内shRNA筛选的功能白血病自我更新的调节剂。基于此屏幕，我们确定了Syncrip，RNA 具有MSI2靶标的结合蛋白在白血病干细胞中需要的结合蛋白（Vu等，2017 自然遗传学）。我们的初步数据，有条件敲除用于合成的数据表明它是也对HSC自我更新至关重要。我们的建议将使我们的重点从MSI2扩大到它相关的监管网络并表征并确定新的分子决定素 HSC和HSPC对称自我更新和不对称命运选择。我们已经改编了新的包括条形码，单细胞RNA-seq和配对女儿HSC分析的技术（命运seq）。我们还开发了一种新的方法来绘制名为HSC中的直接mRNA靶标（HyperTribe），（Nguyen等人自然通讯2020）。该提案将确定新的 HSPC命运选择的监管机构将导致新的治疗策略改善 MDS患者的结果。