Adaptation toex vivoculture drives human haematopoietic stem cell loss of repopulation capacity in a cell cycle independent manner

• DOI: 10.1101/2022.11.17.516906
• Publication Date: 2022-11-18T00:10:17Z
• Authors: Carys S. Johnson; Kendig Sham; Serena Belluschi; Xiaonan Wang; Winnie Lau; Kerstin B. Kaufmann; Gabriela Krivdova; Emily F. Calderbank; Nicole Mende; Jessica McLeod; Giovanna Mantica; Matthew J. Williams; Charlotte Grey-Wi...; Michael Drakopoulos; Shubhankar Sinha; Evangelia Diamanti; Christina Basford; Anthony R. Green; Nicola K. Wilson; Steven J. Howe; John E. Dick; Bertie Göttgens; Natalie Francis; Elisa Laurenti
• Keywords: 0301 basic medicine; Transplantation; 5.2 Cellular and gene therapies; 32 Biomedical and Clinical Sciences; Regenerative Medicine; Stem Cell Research; 03 medical and health sciences; Stem Cell Research - Nonembryonic - Human; FOS: Biological sciences; Genetics; 3206 Medical Biotechnology; Stem Cell Research - Nonembryonic - Non-Human; Biotechnology

AbstractLoss of long-term haematopoietic stem cell function (LT-HSC) hampers the success ofex vivoHSC gene therapy and expansion procedures, but the kinetics and the mechanisms by which this occurs remain incompletely characterized. Here through time-resolved scRNA-Seq, matchedin vivofunctional analysis and the use of a reversiblein vitrosystem of early G1arrest, we define the sequence of transcriptional and functional events occurring during the firstex vivodivision of human LT-HSCs. We demonstrate that contrary to current assumptions, loss of long-term repopulation capacity during culture is independent of cell cycle progression. Instead it is a rapid event that follows an early period of adaptation to culture, characterised by transient gene expression dynamics and constrained global variability in gene expression. Cell cycle progression however contributes to the establishment of differentiation programmes in culture. Our data have important implications for improving HSC gene therapy and expansion protocols.