Mechanisms of human pre-B cell differentiation

人类前B细胞分化的机制

基本信息

批准号：
9243971
负责人：
KATIA GEORGOPOULOS
金额：
$ 20.36万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-15 至 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9243971
关键词：
Adhesions Adult Anoikis B cell differentiation B-Cell Acute Lymphoblastic Leukemia B-Lymphocytes Bone Marrow Cell Adhesion Cell Death Cell surface Chromatin Data Dependence Development Developmental Process Disease Drug resistance Event Fibronectins Gene Expression Gene Expression Profile Gene Mutation Generations Genes Genetic Recombination Genetic Transcription Goals Heavy-Chain Immunoglobulins Hematological Disease Hematopoietic Hematopoietic System Human IL7R gene Immune Immunologic Deficiency Syndromes In Vitro Integrins Investigation Knowledge Ligands Light Lymphoid Malignant - descriptor Malignant Neoplasms Mature B-Lymphocyte Modeling Molecular Mus Mutation Pathway interactions Phase Transition Phenotype Process Property Receptors, Antigen, B-Cell Regulation Role Sampling Signal Pathway Signal Transduction Site Staging Stem cells System Testing Transplantation Umbilical Cord Blood Work cell type differentiated B cell fetal follow-up genome editing high risk human disease in vivo leukemia mutant novel strategies pre-B cell receptor prevent public health relevance self-renewal transcription factor

项目摘要

DESCRIPTION (provided by applicant): Regulated transitions through early B cell differentiation are of great importance for the generation and selection of the mature B cell repertoire. We have recently described a transitional phase in early mouse B cell differentiation that in addition to pre-BCR and IL7R signaling also displays integrin-dependent stromal adhesion, strong proliferative capacity and self-renewal. Loss in the transcriptional regulator Ikaros causes arrest at this stromal- adherent pre-B cell stage with a profound increase in integrin signaling, stromal adhesion and self-renewal and loss in pre-BCR differentiation signaling. Although this stage of early B cell differentiation is also important for the developmen of human hematological disorders, including immunodeficiencies and leukemias, little is known of this process in the human hematopoietic system and is the focus of our current investigation. In the first aim, we will evaluate the hierarchy of human B cell precursors that arise in a bone marrow stromal-dependent system. We will examine the in vitro stromal dependence of early human B cell precursors for proliferative expansion and differentiation and identify the signaling pathways and transcriptional factors associated with these events. The differentiation properties of human B cell precursors generated in vivo in immune-deficient NSG mice will be evaluated in a similar fashion. In the second aim, we will examine the role of IKAROS in human pre-B cell differentiation. The IKZF1 locus will be disrupted by genome editing in human B cell precursors to model mutations in human disease. IKAROS mutant B cell precursors will be tested for differentiation, proliferation and adhesion properties in in vitro stromal cultures and in vivo aftr transplantation in NSG mice. Cellular and molecular data generated from IKAROS deficient human pre-B cell precursors and its normal counterparts can serve as baselines to which hematological disorders can be compared. Together these studies will generate critical new knowledge on the molecular pathways that drive the human developmental process and its aberrant manifestations. This work will enable us to move seamlessly between the mouse and human to discover better approaches for treating hematological disorders.

描述（由申请人提供）：早期 B 细胞分化的调节转变对于成熟 B 细胞库的产生和选择非常重要，我们最近描述了除了前 BCR 之外的早期小鼠 B 细胞分化的过渡阶段。 IL7R 信号传导还表现出整合素依赖性基质粘附、强大的增殖能力和自我更新能力。转录调节因子 Ikaros 的缺失会导致这种基质粘附的前 B 细胞停滞。尽管早期 B 细胞分化的这一阶段对于人类血液疾病（包括免疫缺陷和白血病）的发展也很重要，但其作用却很少。人类造血系统中的这一过程是已知的，也是我们当前研究的重点。第一个目标是，我们将评估骨髓基质依赖性中产生的人类 B 细胞前体的层次结构。我们将检查早期人类 B 细胞前体增殖扩张和分化的体外基质依赖性，并确定与这些事件相关的信号通路和转录因子。在免疫缺陷的 NSG 中体内产生的人类 B 细胞前体的分化特性。将以类似的方式评估小鼠。在第二个目标中，我们将研究 IKAROS 在人类前 B 细胞分化中的作用，通过对人类 B 细胞前体进行基因组编辑来破坏 IKZF1 位点，以模拟人类突变。 IKAROS 突变 B 细胞前体将在体外基质培养物和 NSG 小鼠体内移植后进行分化、增殖和粘附特性测试。作为可以比较血液学疾病的基线，这些研究将产生关于驱动人类发育过程及其异常表现的分子途径的重要新知识。和人类发现治疗血液疾病的更好方法。