Temporal Lysophosphatidic Acid Supplementation Enhances Megakaryocyte Differentiation and Platelet Production From Human Hematopoietic Progenitors

Abstract

Platelet shortages and limited storage stability restrict global platelet transfusion capacity, highlighting the need for efficient in vitro platelet production systems. This study establishes a simple, non-genetic and cost-effective system for efficient in vitro platelet-like particle (PLP) production from human hematopoietic stem/progenitor cells (HSPCs) using a Phase-specific modulation of Hippo-YAP/TAZ signaling modulation. Temporal control of Hippo-YAP/TAZ signaling by lysophosphatidic acid (LPA), an activator of YAP/TAZ activity, significantly enhanced megakaryocyte differentiation, expansion and PLP production, resulting in an approximately 15-fold increase in PLP yield at the end of the procedure. Furthermore, LPA extended the expansion period of HSPC-derived megakaryocytes up to 8 days, resulting in a greater than 20-fold increase in the number of HSPC-derived CD41⁺ megakaryocytes. Moreover, replacement of expensive commercial recombinant human thrombopoietin (C-rhTPO), one of the major cost-driving components in in vitro PLP production, with recombinant human thrombopoietin produced in Escherichia coli (W-rhTPO) further improved the cost-effectiveness of the procedure. In conclusion, this study demonstrates that temporally controlled Hippo-YAP/TAZ signaling, together with affordable cytokine supplementation, provides a robust and GMP-compatible platform for large-scale PLP manufacturing for future clinical applications. We believe that this system will enable scalable PLP generation, even in resource-constrained settings, to increase human platelet supply for many life-saving therapies in the future.