A mitochondrial NADPH-cholesterol axis regulates extracellular vesicle biogenesis to support hematopoietic stem cell fate

Massimo Bonora; Claudia Morganti; Nick van Gastel; Kyoko Ito; Enrica Calura; Ilaria Zanolla; Letizia Ferroni; Yang Zhang; Yookyung Jung; Gabriele Sales; Paolo Martini; Takahisa Nakamura; Francesco Massimo Lasorsa; Toren Finkel; Charles P Lin; Barbara Zavan; Paolo Pinton; Irene Georgakoudi; Chiara Romualdi; David T Scadden; Keisuke Ito

doi:10.1016/j.stem.2024.02.004

A mitochondrial NADPH-cholesterol axis regulates extracellular vesicle biogenesis to support hematopoietic stem cell fate

Cell Stem Cell. 2024 Mar 7;31(3):359-377.e10. doi: 10.1016/j.stem.2024.02.004.

Authors

Massimo Bonora¹, Claudia Morganti¹, Nick van Gastel², Kyoko Ito¹, Enrica Calura³, Ilaria Zanolla⁴, Letizia Ferroni⁵, Yang Zhang⁶, Yookyung Jung⁷, Gabriele Sales³, Paolo Martini⁸, Takahisa Nakamura⁹, Francesco Massimo Lasorsa¹⁰, Toren Finkel¹¹, Charles P Lin¹², Barbara Zavan¹³, Paolo Pinton¹⁴, Irene Georgakoudi⁶, Chiara Romualdi³, David T Scadden¹⁵, Keisuke Ito¹⁶

Affiliations

¹ Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Departments of Oncology and Medicine, Albert Einstein College of Medicine-Montefiore Health System, Bronx, NY 10461, USA.
² Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA; de Duve Institute, UCLouvain, 1200 Brussels, Belgium.
³ Department of Biology, University of Padova, 35121 Padua, Italy.
⁴ Department of Medical Sciences, University of Ferrara, 44121 Ferrara, Italy.
⁵ Maria Cecilia Hospital, GVM Care & Research, Cotignola, 48033 Ravenna, Italy.
⁶ Department of Biomedical Engineering, Tufts University, 4 Colby St, Medford, MA 02155, USA.
⁷ Department of Biomedical Engineering, Tufts University, 4 Colby St, Medford, MA 02155, USA; Department of Pathology, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
⁸ Department of Molecular and Translational Medicine, University of Brescia, 25121 Brescia, Italy.
⁹ Divisions of Endocrinology and Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA; Department of Metabolic Bioregulation, Institute of Development, Aging and Cancer, Tohoku University, Sendai 980-8575, Japan.
¹⁰ Department of Biosciences Biotechnologies and Environment University of Bari and Institute of Biomembranes Bioenergetics and Molecular Biotechnologies, Consiglio Nazionale delle Ricerche, 70125 Bari, Italy.
¹¹ Aging Institute and Department of Medicine, University of Pittsburgh School of Medicine/University of Pittsburgh Medical Center, Pittsburgh, PA 15261, USA.
¹² Center for Systems Biology and Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
¹³ Maria Cecilia Hospital, GVM Care & Research, Cotignola, 48033 Ravenna, Italy; Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, 44121 Ferrara, Italy; Translational Medicine Department, University of Ferrara, 44121 Ferrara, Italy.
¹⁴ Department of Medical Sciences, University of Ferrara, 44121 Ferrara, Italy; Maria Cecilia Hospital, GVM Care & Research, Cotignola, 48033 Ravenna, Italy; Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, 44121 Ferrara, Italy.
¹⁵ Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA.
¹⁶ Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Departments of Oncology and Medicine, Albert Einstein College of Medicine-Montefiore Health System, Bronx, NY 10461, USA; Montefiore Einstein Comprehensive Cancer Center and Diabetes Research Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Electronic address: keisuke.ito@einsteinmed.edu.

PMID: 38458178
PMCID: PMC10957094 (available on 2025-03-07)
DOI: 10.1016/j.stem.2024.02.004

Abstract

Mitochondrial fatty acid oxidation (FAO) is essential for hematopoietic stem cell (HSC) self-renewal; however, the mechanism by which mitochondrial metabolism controls HSC fate remains unknown. Here, we show that within the hematopoietic lineage, HSCs have the largest mitochondrial NADPH pools, which are required for proper HSC cell fate and homeostasis. Bioinformatic analysis of the HSC transcriptome, biochemical assays, and genetic inactivation of FAO all indicate that FAO-generated NADPH fuels cholesterol synthesis in HSCs. Interference with FAO disturbs the segregation of mitochondrial NADPH toward corresponding daughter cells upon single HSC division. Importantly, we have found that the FAO-NADPH-cholesterol axis drives extracellular vesicle (EV) biogenesis and release in HSCs, while inhibition of EV signaling impairs HSC self-renewal. These data reveal the existence of a mitochondrial NADPH-cholesterol axis for EV biogenesis that is required for hematopoietic homeostasis and highlight the non-stochastic nature of HSC fate determination.

Keywords: HSC self-renewal; NADPH; cholesterol; exosomes; extracellular vesicles; fate determination; fatty acid oxidation; hematopoietic stem cell; metabolism; mitochondria.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Cell Differentiation / physiology
Cell Self Renewal
Extracellular Vesicles*
Hematopoietic Stem Cells* / metabolism
NADP / metabolism

Substances

NADP

Abstract

Publication types

MeSH terms

Substances

Grants and funding