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4035 Loss of Cpt1a Results in Elevated Mitochondrial Reactive Oxygen Species from Glucose-Fueled Mitochondrial Oxidative Phosphorylation and Defective Hematopoietic Stem Cells

Program: Oral and Poster Abstracts
Session: 501. Hematopoietic Stem and Progenitor Cells and Hematopoiesis: Basic and Translational: Poster III
Hematology Disease Topics & Pathways:
Research, Fundamental Science
Monday, December 9, 2024, 6:00 PM-8:00 PM

Jue Li1,2,3*, Michihiro Hashimoto, PhD4,5,6*, Mingzhe Pan5,6*, Hui Zeng, MD, PhD7, Paul R. Andreassen, PhD8,9* and Gang Huang, PhD5,6

1Department of Cell Systems and Anatomy, UT Health San Antonio, Joe R. and Teresa Lozano Long School of Medicine, San Antonio, San Antonio, TX
2Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinanti, OH
3Department of Hematology, Xiangya Hospital, Central South University, Changsha, China
4Mays Cancer Center, UT Health San Antonio, San Antonio, TX
5Department of Pathology and Laboratory Medicine, UT Health San Antonio, San Antonio, TX
6Department of Cell Systems and Anatomy, UT Health San Antonio, San Antonio, TX
7Department of Hematology, Guangdong Provincial People's Hospital, South Medical University, Guangzhou, China
8Cincinnati Children's Hosp. Med. Ctr., Cincinnati, OH
9Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati

Background:

Hematopoietic stem cells (HSCs) rely on self-renewal to sustain stem cell potential and undergo differentiation to generate mature blood cells. Mitochondrial fatty acid β-oxidation (FAO) is essential for HSC maintenance. Carnitine palmitoyl transferase 1a (Cpt1a), a key enzyme in FAO, is located in the mitochondrial outer membrane and facilitates the transport of fatty acids into mitochondria, where they are broken down via the FAO pathway, thereby leading to production of ATP. Specifically, the CPT1A-mediated FAO pathway generates NADH, which can be used later by mitochondrial respiratory chain complexes to produce ATP through OXPHOS. Further, acetyl-CoA, an end product of FAO, can enter the tricarboxylic acid cycle (TCA), wherein both NADH and ATP are also generated. Recent studies have highlighted the significant role of mitochondrial metabolism in regulating both the self-renewal and differentiation capacities of HSCs. It is generally believed that a metabolic transition of HSCs from glycolysis to OXPHOS, and the increase of reactive oxygen species (ROS), drives the exit of HSCs from quiescence and initiates their subsequent differentiation. However, the precise role of Cpt1a and FAO in HSCs, and of mitochondrial metabolism more generally, remains elusive.

Results:

Cpt1a is highly enriched in HSCs. In this study, utilizing a Cpt1a hematopoietic specific conditional knock-out (Cpt1aΔ/Δ) mouse model, we found that loss of Cpt1a leads to HSC defects, including loss of HSC quiescence and self-renewal, and increased differentiation. Cpt1aΔ/Δ HSCs have compromised reconstitution and self-renewal capacities. Genotoxic stresses, such as 5-FU and irradiation, downregulate Cpt1a, which restricts glucose-fueled mitochondrial oxidative phosphorylation and mitochondrial ROS production and maintains hematopoietic stem cells. Increased glucose-fueled mitochondrial function in Cpt1aΔ/Δ HSCs. Mechanistically, we found that loss of Cpt1a results in elevated levels of mitochondrial respiratory chain complex components and their activities, as well as increased ATP production, and accumulation of mitochondrial reactive oxygen species (mitoROS) in HSCs. Our data suggests hyperactivation of mitochondria and metabolic rewiring via upregulated glucose-fueled oxidative phosphorylation (OXPHOS).

Conclusion:

In summary, our findings illuminate the intricate connections among genotoxic stresses, Cpt1a, mitochondria and HSCs. Further, Cpt1a emerges as a pivotal player in maintaining HSC homeostasis by modulating mitochondrial function. Finally, our results suggest the possibility that Cpt1a activity and pathway may serve as a potential target for addressing HSC exhaustion and may offer a novel avenue to sustain functional HSCs under various genotoxic stresses and prevent both inherited and acquired bone marrow failures.

Keywords: Cpt1a, LT-HSCs, mitochondrial FAO, glucose-fueled OXPHOS, ROS

Disclosures: Pan: Kind Pharmaceuticals, LLC: Research Funding.

*signifies non-member of ASH