Data-guided direct reprogramming of human fibroblasts into the hematopoietic lineage

Direct reprogramming of human fibroblasts into hematopoietic stem cells (HSCs) presents a promising strategy for overcoming the limitations of traditional bone-marrow transplantation. Despite the potential of this approach, our understanding of the mechanisms driving efficient autologous cell type conversion remains incomplete. Here, we evaluate a novel algorithmically predicted transcription factor (TF) recipe - GATA2, GFIB1, FOS, REL, and STAT5A - for inducing HSC-like states from human dermal fibroblasts. Using flow cytometry and long-read single-cell RNA-sequencing, we demonstrate increased CD34 ⁺ cell populations and high transcriptomic similarity to native HSCs. Additionally, we uncover transcriptional heterogeneity at both gene and isoform levels among induced HSCs, underscoring the complexity of direct reprogramming.