Hypoxia Core

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This revised shared Hypoxia core (HC) is proposed for the Indiana University (IU) Cooperative Center of Excellence in Hematology (CCEH) to support and enhance research efforts of members of our center, other CCEH centers, and other qualified US investigators in the area of hematopoiesis and nonmalignant hematology. Information generated is critical for better in vivo understanding of phenotypic and functional cellular, molecular, and biochemical aspects of hematopoietic stem (HSC), progenitor (HPC), immune, and stromal cells. This involves regulation of normal hematopoiesis and pre-leukemia in context of their lowered oxygen (O2) environment in vivo. O2 tension within bone marrow (BM;1-5%) and cord blood (CB) or mobilized peripheral blood (mPB; <10%), is lower than in the ambient air (~21% O2) in which they are routinely collected for analysis. While it has been known for >40 years that HSC and HPC grow better ex-vivo in lowered (≤ 5%) O2, our report (Mantel, et. al., Cell, 2015) and more recent work shown in this HC grant project, demonstrated previously unknown information that collection/processing of BM, CB, and mPB in ambient air results in large decreases in phenotypically defined and functional HSC and increased HPC numbers within minutes of cell exposure to ambient air. We termed this process Extra Physiological Shock/Stress (EPHOSS). Collection/processing of cells at 3% O2, such that they are never exposed to ambient air resulted in 2 to 5-fold increases in phenotypically- and functionally-detectable HSC. As well, this manifested as different gene expression patterns and responsiveness to stimuli associated with HSC. Similar changes were also noted with murine pre-leukemic and immune cells. Re-evaluation of hematopoietic function associated with maintenance of HSC and HPC at lowered O2 levels can now be elucidated through the expertise of our HC. New data derived with our hypoxia chamber in the revised HC demonstrates the incredible power of the HC to help the hematopoietic community. Specific Aims of the HC involving normal and non-malignant murine and human hematology are: 1) Provide outstanding, consistent and timely analysis of mouse and human BM, mPB, immune and stromal cells, and human CB, BM, and mPB collected/processed at 3%, as well as other, O2 tensions compared to that in ambient air; 2) Coordinate with the other IU CCEH cores for in-depth analysis of cells collected in hypoxia vs. ambient air; 3) Enhance productivity of our CCEH and other CCEH members, and outside investigators by providing a centralized set of services with reduced cost, and expert guidance not currently available elsewhere; and 4) Provide advice regarding intricacies of such studies, analyses, and overcoming potential problems. The HC provides an economic resource that will add new and significant cutting-edge scientific analysis to greatly advance our current understanding of HSC/HPC and immune and stromal cell biology in ways that will increase their translational use for clinical benefit.