pTa-controlled reporter to identify lymphoid precursor

pTa 控制的报告基因识别淋巴前体

• 批准号: 6572039
• 负责人: HARALD VON BOEHMER
• 金额: $ 42.75万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6572039
• 关键词: T cell receptor; T lymphocyte; developmental immunology; gene expression; genetic enhancer element; genetic promoter element; laboratory mouse; lymphopoiesis; reporter genes; transcription factor

DESCRIPTION (provided by applicant): The development of lymphoid lineages from pluripotent stem cells is a poorly understood process. The identification of lymphoid committed precursors is not only of academic but also of clinical interest because of their faster immunorestorative potential when compared to hemopoietic stem cells (HSC) after transplantation into immunodeficient hosts. Previous analysis of the expression of the pre-TCRa (pTa) chain, which forms the pre-TCR in association with the TCRb chain, has shown its presence in precursors of T cells but not in pluripotent hemopoietic stem cells. This suggests that expression of the pTa gene marks lymphoid precursors. Therefore, we propose to isolate lymphoid precursors with the aid of reporter genes that are controlled by pTa promoter and enhancer elements. Our initial data indicate that such reporters can be expressed in lymphoid committed precursors in the bone marrow that are able to generate all lymphoid lineages but no myeloid cells. We propose to subdivide the reporter gene expressing cells by a variety of cell surface molecules that are involved in and correlate with lymphoid development. We will then reveal the developmental potential of these subsets in order to generate a detailed picture of lymphoid commitment in fetal and adult lymphopoiesis and to identify branching points of the various lineages. We will further characterize developmental stages with regard to expression and essential role of transcription factors involved in lineage commitment.

描述（由申请人提供）：从多能干细胞发育出淋巴谱系的过程尚不清楚。淋巴定向前体细胞的鉴定不仅具有学术意义，而且具有临床意义，因为与造血干细胞（HSC）相比，它们在移植到免疫缺陷宿主后具有更快的免疫恢复潜力。先前对前TCRa（pTa）链（与TCRb链结合形成前TCR）表达的分析显示其存在于T细胞前体中，但不存在于多能造血干细胞中。这表明 pTa 基因的表达标志着淋巴前体细胞。因此，我们建议借助 pTa 启动子和增强子元件控制的报告基因来分离淋巴前体细胞。我们的初步数据表明，此类报告基因可以在骨髓中的淋巴定型前体细胞中表达，这些前体细胞能够产生所有淋巴谱系，但不能产生骨髓细胞。我们建议将报告基因表达细胞细分为参与淋巴发育并与之相关的各种细胞表面分子。然后，我们将揭示这些亚群的发育潜力，以便生成胎儿和成人淋巴细胞生成中淋巴样定型的详细图片，并确定各种谱系的分支点。我们将进一步描述谱系定型中涉及的转录因子的表达和重要作用的发育阶段特征。