Biology and function of SALL4-induced hematopoietic stem cells from cord blood

描述(申请人提供)：造血干细胞(HSCs)是一种罕见的细胞，具有自我更新和分化为所有造血系细胞的独特能力。造血干细胞的扩增一直是发展先进的细胞疗法治疗骨髓移植和许多血液疾病的重要目标。在过去的二三十年里，第一批造血生长因子被鉴定出来，人们曾多次尝试使用纯化的生长因子来体外扩增造血干细胞，这些生长因子是已知的调节造血干细胞的生长因子。然而，这些尝试在临床应用中取得的成功有限。研究界迫切需要一种创新的方法来成功地解开HSC扩增生物学，并在体外将HSC扩增到临床有用数量的能力方面取得突破。这将在骨髓移植和血液系统癌症和疾病的基因治疗领域产生巨大影响。为此，我们取得了几项惊人的发现，为使用细胞因子依赖的SALL4技术扩增HSC提供了新的线索，并为本文提出的研究奠定了基础。我们希望我们的研究将发现一种可以开发的全新的HSC扩增形式，因此将这项研究转化为临床环境是可行的。我们提议的具体目标集中在实现这一目标上。目的1.明确SALL4促进HSC体外和体内扩增的动力学、持续时间和幅度。目的2-比较两种不同的扩增人脐血干细胞的方法，并表征短期和长期再扩增对骨髓恢复的贡献程度。目的3.利用肝星状细胞的蛋白质质谱学鉴定与SALL4的N-末端结构域特异相互作用的蛋白质。 公共卫生相关性：造血干细胞(HSCs)是一种罕见的细胞，具有自我更新和分化为所有造血系细胞的独特能力。造血干细胞的扩增一直是发展先进的细胞疗法治疗骨髓移植和许多血液疾病的重要目标。这将在骨髓移植和血液系统癌症和疾病的基因治疗领域产生巨大影响。 个别评审员的书面评论和标准分数以基本上未经编辑的形式在下面的“评论”部分提供。请注意，这些批评意见和标准分数是在会议之前准备的，在审查会议上进行任何讨论后可能没有加以修订。上文“恢复和总结讨论”一节概述了委员会的最后意见。

DESCRIPTION (provided by applicant): Hematopoietic stem cells (HSCs) are rare cells that have the unique ability to self-renew and differentiate into cells of all hematopoietic lineages. The expansion of HSCs has remained an important goal to develop advanced cell therapies for bone marrow transplantation and many blood disorders. During the last two to three decades, in which the first hematopoietic growth factors were identified, there have been numerous attempts to expand HSCs in vitro using purified growth factors that are known to regulate HSCs. However, these attempts have been met with limited success for clinical applications. An innovative approach is urgently needed for the research community to succeed in unraveling HSC expansion biology and creating a breakthrough in the ability to expand HSCs in vitro to clinically useful numbers. This would have tremendous impact in the areas of bone marrow transplantation and gene therapy for hematologic cancers and disorders. Toward this end, we have made several striking discoveries that shed new light on HSC expansion using cytokine-dependent SALL4 technology and lay the groundwork for the studies proposed here. We hope our studies will uncover an entirely new form of HSC expansion that could be developed and it is thus feasible to translate this study into the clinical setting. The Specific Aims of our proposal are focused on achieving this objective. Aim 1. To define the kinetics, duration, and magnitude of SALL4-induced enhancement of HSC expansion ex vivo and in vivo. Aim 2-To compare two different approaches to expand human umbilical cord (hUBC) stem cells and characterize the extent of their contribution of short-term and long-term repopulation to marrow recovery. Aim 3. - To identify proteins that specifically interact with the N-terminal domains of SALL4 by taking advantage of protein mass spectrometry from HSCs. PUBLIC HEALTH RELEVANCE: Hematopoietic stem cells (HSCs) are rare cells that have the unique ability to self-renew and differentiate into cells of all hematopoietic lineages. The expansion of HSCs has remained an important goal to develop advanced cell therapies for bone marrow transplantation and many blood disorders. This would have tremendous impact in the areas of bone marrow transplantation and gene therapy for hematologic cancers and disorders. The written critiques and criteria scores of individual reviewers are provided in essentially unedited form in the "Critique" section below. Please note that these critiques and criteria scores were prepared prior to the meeting and may not have been revised subsequent to any discussions at the review meeting. The "Resume and Summary of Discussion" section above summarizes the final opinions of the committee.