biostudies-literatureUnknown30Bastone ALDeutsche ForschungsgemeinschaftLower Saxony State Ministry of Science and CultureHorizon 2020Safety assessment in retroviral vector-mediated gene therapy remains challenging. In clinical trials for different blood and immune disorders, insertional mutagenesis led to myeloid and lymphoid leukemia. We previously developed the <i>In Vitro</i> Immortalization Assay (IVIM) and Surrogate Assay for Genotoxicity Assessment (SAGA) for pre-clinical genotoxicity prediction of integrating vectors. Murine hematopoietic stem and progenitor cells (mHSPCs) transduced with mutagenic vectors acquire a proliferation advantage under limiting dilution (IVIM) and activate stem cell- and cancer-related transcriptional programs (SAGA). However, both assays present an intrinsic myeloid bias due to culture conditions. To detect lymphoid mutants, we differentiated mHSPCs to mature T cells and analyzed theirMolecular therapy. Methods & clinical development515-533https://www.ebi.ac.uk/biostudies/studies/S-EPMC10491817biostudies-literatureDevelopment of an <i>in vitro</i> genotoxicity assay to detect retroviral vector-induced lymphoid insertional mutants.PMC10491817Dittrich-Breiholz ODziadek VMansel FBastone ALFleischauer JSchwarzer ASchambach AJohn-Neek PAgyeman-Duah ERothe MSchaudien DfalseDevelopment of an <i>in vitro</i> genotoxicity assay to detect retroviral vector-induced lymphoid insertional mutants.Safety assessment in retroviral vector-mediated gene therapy remains challenging. In clinical trials for different blood and immune disorders, insertional mutagenesis led to myeloid and lymphoid leukemia. We previously developed the <i>In Vitro</i> Immortalization Assay (IVIM) and Surrogate Assay for Genotoxicity Assessment (SAGA) for pre-clinical genotoxicity prediction of integrating vectors. Murine hematopoietic stem and progenitor cells (mHSPCs) transduced with mutagenic vectors acquire a proliferation advantage under limiting dilution (IVIM) and activate stem cell- and cancer-related transcriptional programs (SAGA). However, both assays present an intrinsic myeloid bias due to culture conditions. To detect lymphoid mutants, we differentiated mHSPCs to mature T cells and analyzed their2023-01-01T00:00:00Z2023 Sep2026-05-28T15:28:34.679Z2025-04-07T03:29:14.911ZS-EPMC104918173769394910.1016/j.omtm.2023.08.017