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CaMKK2 Knockout Bone Marrow Cells Collected/Processed in Low Oxygen (Physioxia) Suggests CaMKK2 as a Hematopoietic Stem to Progenitor Differentiation Fate Determinant

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Abstract

Little is known about a regulatory role of CaMKK2 for hematopoietic stem (HSC) and progenitor (HPC) cell function. To assess this, we used Camkk2−/− and wild type (WT) control mouse bone marrow (BM) cells. BM cells were collected/processed and compared under hypoxia (3% oxygen; physioxia) vs. ambient air (~21% oxygen). Subjecting cells collected to ambient air, even for a few minutes, causes a stress that we termed Extra Physiological Shock/Stress (EPHOSS) that causes differentiation of HSCs and HPCs. We consider physioxia collection/processing a more relevant way to assess HSC/HPC numbers and function, as the cells remain in an oxygen tension closer physiologic conditions. Camkk2−/− cells collected/processed at 3% oxygen had positive and negative effects respectively on HSCs (by engraftment using competitive transplantation with congenic donor and competitor cells and lethally irradiated congenic recipient mice), and HPCs (by colony forming assays of CFU-GM, BFU-E, and CFU-GEMM) compared to WT cells processed in ambient air. Thus, with cells collected/processed under physioxia, and therefore never exposed and naïve to ambient air conditions, CaMKK2 not only appears to act as an HSC to HPC differentiation fate determinant, but as we found for other intracellular mediators, the Camkk−/− mouse BM cells were relatively resistant to effects of EPHOSS. This information is of potential use for modulation of WT BM HSCs and HPCs for future clinical advantage.

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All data is present in this paper, and material is available upon request.

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Funding

These studies were supported by NIH R35 HL139599 (Outstanding Investigator Award) and U54 DK 106846 (Cooperative Center of Excellence in Hematology (CCEH)) to H.E.B. J.R. was supported as a post-doctoral fellow on T32 DK 007519 to H.E.B. U.S. was supported by R01AR068332 (NIAMS/NIH). LR was supported by NIH 2U19AI067798-16.

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Author notes

  1. Hal E. Broxmeyer, James Ropa and Maegan L. Capitano contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology/Immunology, Indiana University School of Medicine, 950 West Walnut Street, R2 Bldg., Room 302, Indianapolis, IN, 46202-5181, USA

    Hal E. Broxmeyer, James Ropa, Maegan L. Capitano & Scott Cooper

  2. Division of Hematological Malignancies and Cellular Therapy, Duke University Medical Center, Durham, NC, USA

    Luigi Racioppi

  3. Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131, Naples, Italy

    Luigi Racioppi

  4. Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, USA

    Uma Sankar

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  1. Hal E. Broxmeyer
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Contributions

Conceptualization: Hal E. Broxmeyer.

Methodology: James Ropa, Maegan L. Capitano, Scott Cooper, Uma Sankar.

Formal analysis and investigation: Hal E. Broxmeyer, James Ropa, Maegan L. Capitano, Scott Cooper, Uma Sankar.

Writing - original draft preparation: Hal E. Broxmeyer, James Ropa, Maegan L. Capitano, Scott Cooper.

Writing - review and editing: Hal E. Broxmeyer, James Ropa, Maegan L. Capitano, Scott Cooper, Luigi Racioppi, Uma Sankar.

Funding acquisition: Hal E. Broxmeyer, Luigi Racioppi, Uma Sankar.

Resources: Hal E. Broxmeyer, Uma Sankar.

Supervision: Hal E. Broxmeyer.

Corresponding author

Correspondence to James Ropa.

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Broxmeyer, H.E., Ropa, J., Capitano, M.L. et al. CaMKK2 Knockout Bone Marrow Cells Collected/Processed in Low Oxygen (Physioxia) Suggests CaMKK2 as a Hematopoietic Stem to Progenitor Differentiation Fate Determinant. Stem Cell Rev and Rep 18, 2513–2521 (2022). https://doi.org/10.1007/s12015-021-10306-8

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