ena-STUDY-UNIVERSITY OF HAMBURG-14-02-2020-12:32:20:601-346
Genome-wide interference of ZNF423 with B-lineage transcriptional circuitries in acute lymphoblastic leukemia
Aberrant expression of the transcriptional modulator and EBF1-antagonist ZNF423 has been implicated in B-cell leukemogenesis, however its impact on transcriptional circuitries in lymphopoiesis has not been elucidated in a comprehensive manner. Herein, in-silico analyses of multiple expression data sets on 1,354 acute leukemia samples revealed a widespread presence of ZNF423 in various subtypes of acute lymphoblastic leukemia (ALL). Average expression of ZNF423 was highest in ETV6-RUNX1, B-other, and TCF3-PBX1 ALL followed by  BCR-ABL, hyperdiploid and KMT2A-rearranged ALL. In a KMT2A-AFF1 pro-B ALL model, a CRISPR-Cas9-mediated genetic ablation of ZNF423 decreased cell viability and significantly prolonged survival of mice upon xenotransplantation. For the first time, we characterized the genome-wide binding pattern of ZNF423, its impact on the chromatin landscape and differential gene activities in a B-lineage context. In general, chromatin-bound ZNF423 was associated with a depletion of activating histone marks. At the transcriptional level, EBF1-dependent transactivation was disrupted by ZNF423, whereas repressive and pioneering activities of EBF1 were not discernibly impeded. Unexpectedly, we identified an enrichment of ZNF423 at canonical EBF1-binding sites also in the absence of EBF1 indicative of intrinsic EBF1-independent ZNF423 activities. A genome-wide motif search at EBF1-target gene loci revealed that EBF1 and ZNF423 co-regulated genes often contain SMAD1/SMAD4-binding motifs as exemplified by the TGFB1 promoter, which was repressed by ZNF423 outcompeting EBF1 in dependence on its ability to bind EBF1-consensus sites and to interact with EBF1 or SMADs. Overall, these findings underscore the wide scope of ZNF423 activities interfering with B-cell lymphopoiesis and contributing to leukemogenesis.
Aberrant expression of the transcriptional modulator and EBF1-antagonist ZNF423 has been implicated in B-cell leukemogenesis, however its impact on transcriptional circuitries in lymphopoiesis has not been elucidated in a comprehensive manner. Herein, in-silico analyses of multiple expression data sets on 1,354 acute leukemia samples revealed a widespread presence of ZNF423 in various subtypes of acute lymphoblastic leukemia (ALL). Average expression of ZNF423 was highest in ETV6-RUNX1, B-other, and TCF3-PBX1 ALL followed by  BCR-ABL, hyperdiploid and KMT2A-rearranged ALL. In a KMT2A-AFF1 pro-B ALL model, a CRISPR-Cas9-mediated genetic ablation of ZNF423 decreased cell viability and significantly prolonged survival of mice upon xenotransplantation. For the first time, we characterized the genome-wide binding pattern of ZNF423, its impact on the chromatin landscape and differential gene activities in a B-lineage context. In general, chromatin-bound ZNF423 was associated with a depletion of activating histone marks. At the transcriptional level, EBF1-dependent transactivation was disrupted by ZNF423, whereas repressive and pioneering activities of EBF1 were not discernibly impeded. Unexpectedly, we identified an enrichment of ZNF423 at canonical EBF1-binding sites also in the absence of EBF1 indicative of intrinsic EBF1-independent ZNF423 activities. A genome-wide motif search at EBF1-target gene loci revealed that EBF1 and ZNF423 co-regulated genes often contain SMAD1/SMAD4-binding motifs as exemplified by the TGFB1 promoter, which was repressed by ZNF423 outcompeting EBF1 in dependence on its ability to bind EBF1-consensus sites and to interact with EBF1 or SMADs. Overall, these findings underscore the wide scope of ZNF423 activities interfering with B-cell lymphopoiesis and contributing to leukemogenesis.