cMPL-based purification and depletion of human hematopoietic stem cells: implications for pretransplant conditioning

D Daisuke Araki (1Laboratory of Regenerative Therapies for Inherited Blood Disorders, Cellular and Molecular Therapeutics Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, United States) S So Gun Hong (1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States) N Nathaniel Linde (1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States) B Bryan Fisk (3Integrated Data Science Services, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD) N Neelam Redekar C Christi Salisbury-Ruf (1Cellular and Molecular Therapeutics Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD) A Allen Krouse (3Integrated Data Science Services, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD) T Theresa Engels (1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States) J Justin Golomb (1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States) P Pradeep Dagur (5Flow Cytometry Core Facility, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD) S Sumith R. Panicker (6Laboratory of Vascular Thrombosis and Inflammation, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD) Y Yogendra Kanthi (6Laboratory of Vascular Thrombosis and Inflammation, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD) D Diogo M. Magnani Z Zhirui Wang A Andre Larochelle (1Laboratory of Regenerative Therapies for Inherited Blood Disorders, Cellular and Molecular Therapeutics Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, United States)

Abstract

Abstract The thrombopoietin:cMPL signaling axis is a critical regulator of early hematopoiesis. However, the utility of cMPL as a standalone marker for identifying long-term repopulating hematopoietic stem cells (LT-HSCs) within the adult human CD34+ hematopoietic stem and progenitor cell (HSPC) population has not been validated. In this study, we established high cMPL surface expression as a defining feature of human LT-HSCs. Targeting the cMPL receptor facilitated the separation of human LT-HSCs from mature progenitors, a delineation not achievable with c-KIT (CD117). Leveraging this finding, we explored the therapeutic potential of cMPL as a novel target for pretransplant conditioning regimens. We developed a cMPL-targeting immunotoxin and demonstrated its ability to preferentially deplete host cMPLhigh LT-HSCs in murine xenograft models. Evaluation in rhesus macaques confirmed these findings and highlighted a favorable safety profile with rapid systemic clearance within 24 hours of administration. Proof of concept experiments validated the immunotoxin as a novel conditioning agent, enabling donor HSPC engraftment without the use of chemotherapy or irradiation. These findings advance our understanding of the molecular determinants of human hematopoiesis and underscore the potential of cMPL-targeting preparative regimens to improve therapeutic transplantation outcomes.

Article Details

Journal Blood
Volume / Issue Vol. 145, Issue 25
Published June 19, 2025
Pages 2978-2991
ISSN 0006-4971
Publisher Elsevier BV

Journal Info

Blood

Elsevier BV

ISSN: 0006-4971 Health Sciences

Authors (15)

D

Daisuke Araki

1Laboratory of Regenerative Therapies for Inherited Blood Disorders, Cellular and Molecular Therapeutics Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, United States

S

So Gun Hong

1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States

N

Nathaniel Linde

1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States

B

Bryan Fisk

3Integrated Data Science Services, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD

N

Neelam Redekar

C

Christi Salisbury-Ruf

1Cellular and Molecular Therapeutics Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD

A

Allen Krouse

3Integrated Data Science Services, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD

T

Theresa Engels

1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States

J

Justin Golomb

1Translational Stem Cell Biology Branch, NIH/NHLBI, Bethesda, United States

P

Pradeep Dagur

5Flow Cytometry Core Facility, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD

S

Sumith R. Panicker

6Laboratory of Vascular Thrombosis and Inflammation, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD

Y

Yogendra Kanthi

6Laboratory of Vascular Thrombosis and Inflammation, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD

D

Diogo M. Magnani

Z

Zhirui Wang

A

Andre Larochelle

1Laboratory of Regenerative Therapies for Inherited Blood Disorders, Cellular and Molecular Therapeutics Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, United States