Expression profiling of fresh, DMSO or UM171 expanded CD34+ cells by high throughput sequencing
Ex-vivo expansion of hematopoietic stem cells (HSCs) is gaining in importance for cell and gene therapy and requires a shift from dormancy state to activation and cycling. However, abnormal or excessive HSC activation results in reduced self-renewal ability and increased propensity for myeloid-biased differentiation. To date, the threshold of HSC activation that marks “the point of no return” is undetermined. Identifying the molecular determinants that restrain or control HSC activation is essential for preventing accelerated exhaustion of cultured-HSCs. We now report that activation of the E3 ligase complex CRL3KBTBD4 by UM171 not only induces epigenetic changes through CoREST1 degradation but also control chromatin bound MYC levels to prevent excessive activation and maintain lympho-myeloid potential of expanded populations. Furthermore, reconstitution activity and multipotency of UM171-treated HSC is specifically compromised when MYC levels are experimentally increased despite degradation of CoREST1. Overall design: Expression profiling of fresh, DMSO or UM171 expanded CD34+ cells by high throughput sequencing