Transmembrane TNF-TNFR2 axis promotes generation of human T cell precursors.

The goal of this study is to dissect the impact of soluble (sTNF) and transmembrane (tmTNF) forms of tumour necrosis factor on the differentiation of human hematopoietic stem and progenitor cells (HSPCs) into T cell precursors. To achieve this, we utilized scRNA-sequencing technologies to compare the transcriptome profiles of HSPCs that had been differentiated in the artificial thymic organoid (ATO) system in the absence (control) or presence of TNF signal (soluble or transmembrane). Overall design: CD3-CD14-CD19-CD56-CD34+ umbilical cord blood HSPCs from 4 donors were pooled and induced to undergo T cell development using the ATO system. In the absence of TNF signal (control), ATOs were supplemented with interleukin-7 and FMS-like tyrosine kinase 3 ligand. For sTNF-activated ATOs, organoids were supplemented additionally with 0.25 ng/mL of sTNF. For tmTNF-activated ATOs, 1 % of the stromal cells that were used to assemble organoids were engineered to express tmTNF. At day 10 post culture, CD45+ differentiating HSPCs were sorted from all three conditions and analysed using scRNA-seq.