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A humanized bone marrow ossicle xenotransplantation model enables improved engraftment of healthy and leukemic human hematopoietic cells

Nature Medicine volume 22, pages 812–821 (2016)Cite this article

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Abstract

Xenotransplantation models represent powerful tools for the investigation of healthy and malignant human hematopoiesis. However, current models do not fully mimic the components of the human bone marrow (BM) microenvironment, and they enable only limited engraftment of samples from some human malignancies. Here we show that a xenotransplantation model bearing subcutaneous humanized ossicles with an accessible BM microenvironment, formed by in situ differentiation of human BM-derived mesenchymal stromal cells, enables the robust engraftment of healthy human hematopoietic stem and progenitor cells, as well as primary acute myeloid leukemia (AML) samples, at levels much greater than those in unmanipulated mice. Direct intraossicle transplantation accelerated engraftment and resulted in the detection of substantially higher leukemia-initiating cell (LIC) frequencies. We also observed robust engraftment of acute promyelocytic leukemia (APL) and myelofibrosis (MF) samples, and identified LICs in these malignancies. This humanized ossicle xenotransplantation approach provides a system for modeling a wide variety of human hematological diseases.

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Figure 1: Human HSPCs and primary acute leukemia engraft robustly in human BM-MSC-derived ossicles.
Figure 2: Primary AML blasts preferentially engraft humanized ossicle niches, as compared to mouse BM.
Figure 3: Humanized ossicle niche transplantation reveals an increased LIC frequency in AML.
Figure 4: Humanized ossicle niches facilitate robust engraftment of APL cells, which are probably derived from lineage-committed LICs.
Figure 5: Humanized ossicle niches facilitate robust engraftment of MF, which derives from LICs in the HSC compartment.

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Acknowledgements

We acknowledge the Hematology Division Tissue Bank and the patients for donating their samples. We acknowledge F. Zhao for lab management; N. Hofmann and B. Luo for technical assistance with ossicle analysis and calreticulin sequencing; and the Stanford Cytogenetics Lab for FISH analysis. A.R. is supported by an Erwin-Schroedinger Research Fellowship (Austrian Science Fund) and D.T. by a CJ Martin Overseas Biomedical Research Fellowship (NHMRC, Australia). R.M. is a New York Stem Cell Foundation Robertson Investigator. This research was supported by the New York Stem Cell Foundation and National Institutes of Health grants R01CA188055 and U01HL099999 to R.M. and by funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No. 668724 to D.S.

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Authors and Affiliations

  1. Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, California, USA

    Andreas Reinisch, Daniel Thomas, M Ryan Corces, Xiaohua Zhang, Wan-Jen Hong & Ravindra Majeti

  2. Cancer Institute, Stanford University School of Medicine, Stanford, California, USA

    Andreas Reinisch, Daniel Thomas, M Ryan Corces, Xiaohua Zhang, Wan-Jen Hong & Ravindra Majeti

  3. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA

    Andreas Reinisch, Daniel Thomas, M Ryan Corces, Xiaohua Zhang, Wan-Jen Hong & Ravindra Majeti

  4. Department of Pathology, Stanford University School of Medicine, Stanford, California, USA

    Dita Gratzinger

  5. Department of Blood Group Serology and Transfusion Medicine, Paracelsus Medical University, Salzburg, Austria

    Katharina Schallmoser

  6. Spinal Cord Injury and Tissue Regeneration Center Salzburg, Paracelsus Medical University, Salzburg, Austria

    Katharina Schallmoser & Dirk Strunk

  7. Experimental and Clinical Cell Therapy Institute, Paracelsus Medical University, Salzburg, Austria

    Dirk Strunk

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Contributions

A.R. and R.M conceived and designed the project. A.R., D.T., M.R.C., W.-J.H., and X.Z. performed the experimental work. A.R., D.T., M.R.C., and D.G. analyzed the data. K.S. and D.S. provided crucial reagents. A.R. and R.M. wrote the manuscript. All authors discussed the results.

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Correspondence to Ravindra Majeti.

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Reinisch, A., Thomas, D., Corces, M. et al. A humanized bone marrow ossicle xenotransplantation model enables improved engraftment of healthy and leukemic human hematopoietic cells. Nat Med 22, 812–821 (2016). https://doi.org/10.1038/nm.4103

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