Glutathione Protects Hematopoietic Stem Cells from Proteotoxic Stress

Glutathione is necessary for maintaining proteostasis in hematopoietic stem cells but may not be crucial for preventing ferroptosis.

Summary and findings

This study investigated the role of glutathione in hematopoietic stem cells (HSCs) by examining the effects of deleting glutamate-cysteine ligase catalytic subunit (GCLC) and glutathione peroxidase 4 (GPX4). GCLC deficiency eliminated glutathione from hematopoietic cells, severely impairing HSC function and hematopoiesis. The findings suggest that glutathione is necessary for proteostasis in HSCs.

Abstract

Glutathione is a redox buffer, best known for preventing oxidative stress and ferroptosis, but rarely studied in stem cells. We studied glutathione function in hematopoietic stem cells (HSCs) by deleting the glutamate-cysteine ligase catalytic subunit (GCLC), which synthesizes glutathione, or glutathione peroxidase 4 (GPX4), which employs glutathione to prevent ferroptosis. GCLC deficiency eliminated glutathione from hematopoietic cells, severely impairing HSC function and hematopoiesis. In contrast, GPX4 deficiency did not affect HSC function and had little effect on hematopoiesis, even after vitamin E depletion, suggesting glutathione has ferroptosis-independent functions in HSCs. Consistent with this, GCLC deficiency increased reactive oxygen species levels in hematopoietic cells. But treatment with antioxidants or ferroptosis inhibitors gave little or no rescue of HSC function despite reducing ROS levels. Rather, GCLC-deficient, but not GPX4-deficient, HSCs exhibited unfolded protein accumulation, proteostatic stress, and cell death marked by elevated p53 levels and Annexin V staining. Deficiency for p53 prolonged the function of GCLC-deficient HSCs after transplantation. Glutathione is thus necessary in HSCs for proteostasis but may be dispensable for ferroptosis inhibition in vivo.

Limitations

Not reported in abstract.

Elsewhere in the Glutathione (GSH) corpus