Preliminary transcriptome profiling of induced pluripotent stem cell-derived hematopoietic stem and progenitor cells in Kawasaki disease

Abstract

The etiology of Kawasaki disease (KD) remains unclear. Hematopoietic stem and progenitor cells (HSPCs) serve as the precursor cells for a multitude of immune cells. Investigating their initial transcriptional status may help uncover the aberrant immune mechanisms underlying KD. Induced pluripotent stem cells (iPSCs) were reprogrammed from peripheral blood mononuclear cells (PBMCs) isolated from patients with KD and febrile individuals, followed by directed differentiation into HSPCs. We performed bulk RNA-sequencing to compare transcriptomic profiles of iPSC-derived HSPCs between the two groups, with further comparison against integrated HSPC data from public KD single-cell datasets. We recruited three patients with KD prior to Intravenous immunoglobulin (IVIG) therapy and three febrile control patients, and successfully established an iPSC-derived HSPCs disease model. Transcriptomic profiling revealed elevated immune and inflammatory response signatures in iPSC-derived HSPCs from patients with KD compared with those from febrile individuals. In addition, in vitro -generated KD iPSC-HSPCs exhibited partial transcriptional features similar to in vivo HSPCs isolated from PBMCs of patients with KD. Gene Set Enrichment Analysis (GSEA) further revealed that gene sets associated with B-cell developmental processes were transcriptionally downregulated in iPSC-derived HSPCs from patients with KD relative to febrile controls. iPSC-derived HSPCs from patients with KD display altered immune-inflammatory transcriptional profiles and suppressed B-cell developmental signatures at the transcriptomic level. These preliminary findings suggest that early hematopoietic immune dysregulation may contribute to KD pathogenesis. We propose that these iPSC-derived HSPCs could be a good cellular model for studying the etiology of KD in vitro . These findings are preliminary, constrained by the small sample size and limited to transcriptomic analysis only. Further studies with larger cohorts and functional experiments are needed to verify these results.