Elevated 5-HTR7 deteriorates dysregulated megakaryocytopoiesis in immune thrombocytopenic purpura via up-regulating the PKA/Orai1/ERK1/2 pathway.

Abstract

Dysregulated megakaryocytopoiesis contributes to reduced platelet counts in immune thrombocytopenic purpura (ITP), yet the mechanism remains elusive. Although 5-hydroxytryptamine receptor 7 (5-HTR7) has been implicated in megakaryocyte (MK) biology, its pathogenic involvement in ITP is undefined. This study investigated the impact of 5-HTR7 on MK maturation in ITP using flow cytometry, immunofluorescence, and single-cell RNA sequencing (scRNA-seq). Analyses revealed elevated 5-HTR7 expression on MKs from ITP patients compared to healthy controls. Pharmacological inhibition of 5-HTR7 using SB269970A not only rescued MK maturation defects in vitro but also restored circulating platelet levels in a mouse model of active ITP. scRNA-seq coupled with western blot validation identified ERK1/2 phosphorylation in SB269970A-treated MKs. Mechanistically, 5-HTR7 impaired MK maturation through the PKA/Orai1/ERK axis by suppressing store-operated calcium entry (SOCE), as confirmed via confocal microscopy. In conclusion, elevated expression of 5-HTR7 impairs maturation of MKs causing lower platelet count in ITP, offering a potential therapeutic target for ITP management.