From Mechanisms of Leukaemogenic Dysregulation to New Candidate Therapeutics
type: CreativeWork
description: The maintenance of a continuous flow of mature blood cells from a reservoir of specialised haematopoietic stem cells requires regulation from an intricate network of regulatory programmes. Gene regulatory networks are well evolved to control multi-lineage commitment and differentiation while maintaining self-renewal capacity in haematopoietic stem and progenitor cells. In leukaemia, mutations commonly hijack these regulatory programs causing both a block in normal differentiation and the misplacement of self-renewal capabilities. Rearrangements involving the *MLL1* gene (*Kmt2a*) produce powerful oncogenic proteins occurring predominantly in paediatric and infant acute myeloid and lymphoid leukaemias. Despite having more than 80 fusion partners, MLL-AF9, MLL-AF4 and MLL-ENL make up more than 60% of cases involving *MLL1* translocations. Conventional retroviral transduction models commonly use lineage depleted or c-Kit enriched mouse bone marrow haematopoietic stem and progenitor cells (HSPCs) that display distinct heterogeneity when profiled at the single-cell level which could explain inconsistencies across previously reported models. Conversely, our model, referred to as ME-Transformed cells, implements a conditionally blocked multipotent haematopoietic progenitor murine cell line (HoxB8-FL) allowing for the reproducible tracking of early leukaemic transformation and transcriptional changes due to MLL-ENL expression (Basillico., *et al* 2020). Using ME-Transformed cells, we knocked out 64 genes via CRISPR-Cas9 followed by small-bulk RNA-Sequencing. The 64 targets include transcription factors (TFs) and key transcriptional regulators implicated in the aetiology of MLL rearranged leukaemia. Bioinformatic analysis revealed a regulatory network connecting 19,437 links across 7,548 genes. Previous work in the Göttgens lab used a similar approach to establish the gene regulatory function of 39 TFs in the wild-type parental HoxB8-FL cells (Kucinski *et al*. 2020). The fi…
abstract: The maintenance of a continuous flow of mature blood cells from a reservoir of specialised haematopoietic stem cells requires regulation from an intricate network of regulatory programmes. Gene regulatory networks are well evolved to control multi-lineage commitment and differentiation while maintaining self-renewal capacity in haematopoietic stem and progenitor cells. In leukaemia, mutations commonly hijack these regulatory programs causing both a block in normal differentiation and the misplacement of self-renewal capabilities. Rearrangements involving the *MLL1* gene (*Kmt2a*) produce powerful oncogenic proteins occurring predominantly in paediatric and infant acute myeloid and lymphoid leukaemias. Despite having more than 80 fusion partners, MLL-AF9, MLL-AF4 and MLL-ENL make up more than 60% of cases involving *MLL1* translocations. Conventional retroviral transduction models commonly use lineage depleted or c-Kit enriched mouse bone marrow haematopoietic stem and progenitor cells (HSPCs) that display distinct heterogeneity when profiled at the single-cell level which could explain inconsistencies across previously reported models. Conversely, our model, referred to as ME-Transformed cells, implements a conditionally blocked multipotent haematopoietic progenitor murine cell line (HoxB8-FL) allowing for the reproducible tracking of early leukaemic transformation and transcriptional changes due to MLL-ENL expression (Basillico., *et al* 2020). Using ME-Transformed cells, we knocked out 64 genes via CRISPR-Cas9 followed by small-bulk RNA-Sequencing. The 64 targets include transcription factors (TFs) and key transcriptional regulators implicated in the aetiology of MLL rearranged leukaemia. Bioinformatic analysis revealed a regulatory network connecting 19,437 links across 7,548 genes. Previous work in the Göttgens lab used a similar approach to establish the gene regulatory function of 39 TFs in the wild-type parental HoxB8-FL cells (Kucinski *et al*. 2020). The fi…
author: Kennedy, Alison
language: eng
keywords: leukaemia, MLL-ENL, network, therapeutics, transcription factor
license: https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/3eacdcc6-4b85-4ed9-b449-a01b6760a0b3/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2023-09-01
date modified: 2026-07-22T22:24:31Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.107034