Controlling Influences of Oxygen Tension and CD26/DPP4 Enzymatic Activity on Regulation of Hematopoietic Stem/Progenitor Cells and Hematopoiesis During Health, Ageing, and Disease

虽然对造血干细胞(HSC)和祖细胞(HPC)的表型(细胞表面和 细胞内标记物)、异质性和功能，仍有更多需要了解，特别是如何最好地 使用和操纵这些细胞以获得治疗优势。PI已经连续运行了由NIH资助的基础 拥有超过39年的科学实验室经验。他的目标仍然是将机械论的洞察力转化为临床前 细胞和动物模型，并与临床研究人员合作，以实现临床实用。美国国立卫生研究院的目的是 R35的应用是为了让PI和他的实验室有充足的不间断时间来追求新想法和创造力 在两个现行的NHLBI R01赠款的背景下的方法。HL 056416侧重于氧分压、代谢、 骨髓(BM)和脐带血(CB)中HSCs的功能及其最佳采集和活性 对其他类型的干细胞和祖细胞也有影响。小鼠骨髓细胞或人脐血细胞暴露于 几分钟内的环境空气会引起额外的生理性休克应激(EPHOSS)，通过 活性氧(ROS)诱导分化，而不是细胞死亡。EPHOSS可能会对 个性化医疗以更准确地评估基因表达谱和细胞反应性 与细胞相比，在体内微环境中与细胞氧分压关系更密切的情况 在环境空气的人工环境中收集/处理。HL 112669评估以下各项的影响 CD26/DPP 4在截断选定细胞因子(如GM-CSF、G-CSF、IL-3、EPO、TPO)中的作用 和趋化因子(如CXCL12、CCL3)，从而改变它们的受体结合、细胞和细胞内活性。 这与细胞因子/趋化因子发挥调节作用的所有系统都有关系。阻止EPHOSS或 抑制CD26/DPP4酶活性会导致HSC数量增加。我们将生产 关于造血机制调控的新信息。我们的假设和意义是： 了解DPP4的EPHOSS和酶活性及其调控将有助于更好地 了解HSC/HPC在健康、衰老和疾病中的作用，信息可用于 加速患者从应激/疾病和人脐血造血细胞移植(HCT)中恢复。 我们的目标是确定：1)什么低氧收集、加工、细胞培养和体内植入，以及 DPP4抑制，告知我们健康期间HSCs、HPC和造血的调节，作为回应 应激、衰老和疾病；2)其他补偿EPHOSS影响的手段，以提高HCT的效率； (3)HSC和HPC细胞周期状态及基因和蛋白质表达谱的差异 在低氧/空气和/或-/DPP4抑制下收集/处理。创新之处在于范式转换 要用最先进的方法解决的概念。我们发现的东西可以翻译到其他细胞上 细胞移植和再生医学新兴领域的细胞/程序的来源，以加强治疗 功效。

While much is known about hematopoietic stem (HSC) and progenitor cell (HPC) phenotype (cell surface and intracellular markers), heterogeneity, and function, there is still more to be learned, especially for how best to use and manipulate these cells for therapeutic advantage. The PI has run a continuously NIH-funded basic science laboratory for over 39 years. His goal continues to be to translate mechanistic insight for pre-clinical cell and animal models, and in collaboration with clinical investigators for clinical utility. The purpose of this NIH R35 application is to allow the PI and his lab ample uninterrupted time to pursue new ideas and creative approaches in context of two active NHLBI R01 grants. HL 056416 focuses on oxygen tension, metabolism, function and optimal collection and activities of HSCs from bone marrow (BM) and cord blood (CB), findings with implications for other stem and progenitor cell types. Exposure of mouse BM or human CB cells to ambient air within minutes induces Extra Physiological Shock Stress (EPHOSS), causing loss of HSCs through Reactive Oxygen Species (ROS) induced differentiation, not cell death. EPHOSS may have implications for personalized medicine to more accurately evaluate gene expression profiles and cell responsiveness in situations more closely related to O2 tension of cells in their in vivo microenvironment, compared to cells collected/processed in an artificial environment of ambient air. HL 112669 assesses effects of CD26/Dipeptidylpeptidase (DPP)4 in truncating selected cytokines (e.g. GM-CSF, G-CSF, IL-3, EPO, TPO) and chemokines (e.g. CXCL12, CCL3), thus changing their receptor-binding, cellular and intracellular activities. This has relevance to all systems in which cytokines/chemokines play a regulatory role. Blocking EPHOSS or inhibiting CD26/DPP4 enzymatic activity results in collection of increased numbers of HSCs. We will produce new information on mechanistic regulation of hematopoiesis. Our hypotheses and significance are: understanding EPHOSS and enzymatic activities of DPP4 and their manipulation will lead to a better understanding of HSC/HPC functions in health, ageing, and disease, information that can be used to accelerate patient recovery from stress/disease and for human CB hematopoietic cell transplantation (HCT). Our goals are to determine: 1) what hypoxic collection, processing, cell culture, and in vivo engraftment, and DPP4 inhibition, informs us about the regulation of HSCs, HPCs, and hematopoiesis during health, in response to stress, ageing and disease; 2) other means to compensate for effects of EPHOSS for more efficient HCT, and 3) differences in cell cycle status and gene and protein expression profile of HSCs and HPCs collected/processed in hypoxia/air and/or -/+ DPP4 inhibition. The innovation lies in paradigm changing concepts to be addressed with state-of-the-art methodologies. What we find can be translated to other cell sources for HCT, and for cells/procedures in the emerging field of regenerative medicine to enhance treatment efficacy.