Session: 501. Hematopoietic Stem and Progenitor Cells and Hematopoiesis: Basic and Translational: Poster I
Hematology Disease Topics & Pathways:
Fundamental Science, Research, Hematopoiesis, Diseases, Immune Disorders, Immunodeficiency, Neutropenia, White blood cell disorders, Metabolism, Biological Processes, Molecular biology
To assess the mechanistic basis of RD, we performed single-cell RNA-sequencing on bone marrow samples of two previously reported patients with biallelic AK2 c.542G>A, p.R175Q missense mutations, and nine healthy donor controls. A gene set enrichment analysis revealed that AK2 deficiency had opposing effects on the granulocytic and lymphoid lineages. While anabolic pathways related to ribonucleoprotein synthesis were paradoxically upregulated in granulocytes from RD patients, the same pathways were profoundly downregulated in lymphoid cells from RD patients relative to healthy donors.
To further investigate how different hematopoietic lineages respond to metabolic stress caused by AK2 deficiency, we have developed an inducible Ak2 knockout mouse model under the control of Mx1-Cre that specifically targets hematopoietic cells (Ak2fl/fl; Mx1-Cre). We found that the consequences of Ak2 deficiency for the hematopoietic system are contingent on the effective engagement of metabolic checkpoints. In hematopoietic stem and progenitor cells, Ak2 deficiency mildly reduced mechanistic target of rapamycin (mTOR) signaling and anabolic pathway activation. This conserved nutrient homeostasis and maintained cell survival. Later during granulopoiesis, metabolic checkpoints became ineffective, leading to a paradoxical upregulation of mTOR activity and energy-consuming anabolic pathways. This caused nucleotide imbalance and the depletion of essential substrates, ultimately resulting in proliferation arrest and demise of the granulocyte lineage. In developing T cells (thymocytes), however, mTOR activity and anabolic pathways remain profoundly suppressed, and Ak2-deficient thymocytes failed to mature past the double-positive (DP) stage. Notably, T cell receptor (TCR) positive cells were completely absent in Ak2-deficient thymocytes, suggesting that Ak2 activity is indispensable for TCR rearrangement. Our findings suggest that although intricate metabolic checkpoint control may help tolerate severe metabolic defects, the failure or overactivity of these checkpoints, as observed in granulocytic and T lineages, both result in a catastrophic loss of metabolic homeostasis and failure of maturation. Our ongoing work characterizes the essential role of AK2 during intra-thymic T cell development.
Disclosures: No relevant conflicts of interest to declare.