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EBF1 and PAX5 control pro-B cell expansion via opposing regulation of the Myc gene

Somasundaram, Rajesh and Jensen, Christina T. and Tingvall-Gustafsson, Johanna and Åhsberg, Josefine and Okuyama, Kazuki and Prasad, Mahadesh and Hagman, James and Wang, Xun and Soneji, Shamit and Strid, Tobias and Ungerbäck, Jonas and Sigvardsson, Mikael (2021) EBF1 and PAX5 control pro-B cell expansion via opposing regulation of the Myc gene. Blood . ISSN 0006-4971. (In Press) https://resolver.caltech.edu/CaltechAUTHORS:20210224-125024033

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Abstract

Genes encoding B lineage restricted transcription factors are frequently mutated in B-lymphoid leukemias, suggesting a close link between normal and malignant B-cell development. One of these transcription factors is Early B cell Factor 1 (EBF1), a protein of critical importance for lineage specification and survival of B-lymphoid progenitors. Here, we report that impaired EBF1 function in mouse B-cell progenitors results in reduced expression of Myc. Ectopic expression of MYC partially rescued B-cell expansion in the absence of EBF1 both in vivo and in vitro. Using chromosome conformation analysis in combination with ATAC-seq, ChIP-seq and reporter gene assays, we identified six EBF responsive enhancer elements within the Myc locus. CRISPR-Cas9 mediated targeting of EBF1 binding sites identified one element of key importance for Myc expression and pro-B cell expansion. These data provide evidence that Myc is a direct target of EBF1. Furthermore, ChIP-seq analysis revealed that several regulatory elements in the Myc locus are targets of PAX5. However, ectopic expression of PAX5 in EBF1 deficient cells inhibits the cell cycle and reduces Myc expression, suggesting that EBF1 and PAX5 act in an opposing manner to regulate Myc levels. This hypothesis is further substantiated by the finding that Pax5 inactivation reduces requirements for EBF1 in pro-B cell expansion. The binding of EBF1 and PAX5 to regulatory elements in the human MYC gene in a B-ALL cell line indicate that the EBF1:PAX5:MYC regulatory loop is conserved and may control both normal and malignant B-cell development.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1182/blood.2020009564DOIArticle
ORCID:
AuthorORCID
Tingvall-Gustafsson, Johanna0000-0002-7359-6584
Hagman, James0000-0002-5436-8455
Wang, Xun0000-0002-1653-5750
Strid, Tobias0000-0002-2166-5170
Ungerbäck, Jonas0000-0002-2190-3896
Sigvardsson, Mikael0000-0001-8527-7276
Alternate Title:Opposing regulation of Myc by EBF1 and PAX5
Additional Information:© 2021 American Society of Hematology. We are grateful for the technical assistance provided by Liselotte Lenner, Linda Bergström and Maria Malmberg. This work was supported by grants from the Swedish Cancer Society (2017-258), the Swedish Childhood Cancer Foundation (2019-0020), the Swedish Research Council (2018-02448), including a Strategic research grant to Stem Therapy, Knut and Alice Wallenberg’s Foundation (2014-0089) and a donation from Henry Hallberg (all above to MS) and Lions forskningsfond mot folksjukdomar (to TS). JH is funded by NIH grant R21AI115696 and by the Wendy Siegel Fund for Leukemia and Cancer Research. Author contributions: KO, TS, RS, CJ, JTG, MP, JÅ, MS and JU designed conducted and analyzed experiments. SS, JU, CJ and TS contributed to the bioinformatic analysis. JRH and XW contributed essential reagents and analyzed data. MS designed experiments analyzed data and wrote the manuscript draft. All authors contributed to the final version of the manuscript. None of the authors has a relevant conflict of interest.
Funders:
Funding AgencyGrant Number
Swedish Cancer Society2017-258
Swedish Childhood Cancer Foundation2019-0020
Swedish Research Council2018-02448
Stem TherapyUNSPECIFIED
Knut and Alice Wallenberg Foundation2014-0089
Henry HallbergUNSPECIFIED
Lions forskningsfond mot folksjukdomarUNSPECIFIED
NIHR21AI115696
Wendy Siegel Fund for Leukemia and Cancer ResearchUNSPECIFIED
Subject Keywords:atac trial, b-lymphocytes, burkitt's lymphoma, cell growth, chromatin immunoprecipitation sequencing, c-myc genes, crispr-cas9, fibrinogen, genes, myc, leukemia
Record Number:CaltechAUTHORS:20210224-125024033
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210224-125024033
Official Citation:Rajesh Somasundaram, Christina T Jensen, Johanna Tingvall-Gustafsson, Josefine Åhsberg, Kazuki Okuyama, Mahadesh Prasad, James Hagman, Xun Wang, Shamit Soneji, Tobias Strid, Jonas Ungerbäck, Mikael Sigvardsson; EBF1 and PAX5 control pro-B cell expansion via opposing regulation of the Myc gene. Blood 2021; blood.2020009564. doi: https://doi.org/10.1182/blood.2020009564
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:108175
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:24 Feb 2021 21:07
Last Modified:24 Feb 2021 21:07

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