Plcl1 Regulates Hematopoietic Stem Cell Function During Aging and Stress by Modulating Calcium Dynamics

Abstract

Long-term hematopoietic stem cells (HSCs) possess the capacity to generate all blood lineages, yet they normally remain quiescent and become activated only in response to acute stress. We previously demonstrated that quiescent HSCs exhibit heterogeneity in intracellular calcium; however the mechanisms that establish this heterogeneity and its physiological relevance remain unclear. Here, we identify phospholipase C like 1 (Plcl1), a non-catalytic protein that binds inositol 1,4,5-trisphosphate (IP), as being specifically enriched in the most quiescent HSC subset. Loss-of-function studies demonstrated that, at steady state, Plcl1 deficiency lowers basal intracellular calcium and shifts the HSC compartment toward CD41 subsets, while preserving overall HSC numbers and long-term reconstitution capacity. Under acute hematopoietic stress, Plcl1 loss accelerated and amplified platelet rebound and expansion of non-canonical megakaryocyte progenitors (ncMkPs), suggestive of activation of the thrombopoietic bypass pathway. In aging HSCs, Plcl1 deficiency exacerbated hallmark features, such as expansion of the phenotypic HSC pool, accumulation of CD41 HSCs and ncMkPs, and myeloid-skewed differentiation with impaired competitive reconstitution. These alterations were accompanied by reduced induction of calcium-responsive immediate-early genes. Collectively, these findings identify Plcl1 as an intrinsic regulator that stabilizes cytosolic calcium dynamics in HSCs, thereby restraining stress- and aging-associated megakaryocytic priming and preserving HSC function.