- Open Access
Wdr5 is essential for fetal erythropoiesis and hematopoiesis
Experimental Hematology & Oncology volume 12, Article number: 39 (2023)
WDR5 is a highly conserved protein that performs multiple scaffolding functions in the context of chromatin. However, efforts to understand the function of WDR5 in normal tissues physiologically are quite limited so far. In our study, we explored the function of Wdr5 in erythropoiesis and hematopoiesis by using a hematopoietic-specific Wdr5 knockout mouse model. We found that loss of Wdr5 mediated by Vav-iCre leads to embryonic lethality with defective erythropoiesis. In addition, Wdr5-deficiency completely impairs the hematopoietic stem and progenitor cells function and might alter the immunophenotype of these stem cells and progenitors by decreasing c-Kit expression. Collectively, we identified the pivotal role of Wdr5 in fetal hematopoiesis and erythropoiesis as the de novo findings.
To the editor,
The canonical function of WDR5 is as a core component of the MLL histone methyltransferase complexes . While the function of several other subunits of MLL complexes has been well elucidated in hematopoiesis [2,3,4,5,6,7,8,9,10,11,12], little is known about the function of WDR5. Our study aimed to identify the role of Wdr5 in normal hematopoiesis by utilizing hematopoietic lineage specific knockout mouse model.
To investigate the function of Wdr5 in normal hematopoiesis, Wdr5 was conditionally deleted in the hematopoietic lineage by Vav-iCre transgenic mice (Additional file 1: Fig. S1A). Strikingly, we did not observe any Wdr5f/f, Vav-iCre mice (referred as CKO mice) among 40 offspring from the intercross between the male Wdr5f/f mice and the female Wdr5f/+, Vav-iCre mice at weaning age (Fig. 1A), which urged us to further dissect the effects at the fetal stage. Interestingly, the CKO embryos showed roughly normal morphology, but much paler body color and smaller-sized fetal livers (FLs) compared with the littermate control at E15.5 and E16.5 (Fig. 1B and Additional file 1: Fig. S1B). Consistently, the total cell number of FL from CKO was robustly decreased at E15.5 (Fig. 1C). We further dissected this at E13.5, and similar effect was observed (Additional file 1: Fig. S1C). However, the absolute cell numbers of FLs were comparable between CKO and the littermate control embryos at E12.5 (Additional file 1: Fig. S1D). These indicated that the defective erythropoiesis might occur in Wdr5-deficient embryos. Next, we evaluated the erythropoiesis (Additional file 1: Fig. S1E). Compared with the littermate control, CKO embryos showed an increased percentage of the immature population including S0-S2 but a decreased percentage of relative mature population S4, whereas the absolute cell numbers of all stages were reduced at E15.5 (Fig. 1D and Additional file 1: Fig. S1F). In addition, Ter119+ cells could be further divided into EryA/B/C based on the FSC parameter (Additional file 1: Fig. 1E). Though the absolute cell numbers of EryA/B/C were reduced, the percentage of EryA was increased and the percentage of EryB/C was decreased in CKO embryos at E15.5 (Fig. 1E and Additional file 1: Fig. S1F). Furthermore, we examined whether the terminal erythroid differentiation is impaired in Wdr5-deficient embryos by Hoechst 33342 staining. Although the percentage of the enucleated erythrocytes were comparable in CKO and the littermate control embryos, the total number of the enucleated erythrocytes was decreased robustly in CKO embryos (Fig. 1F) and the enucleated cells from CKO embryos showed a larger size compared with those from the littermate control determined by the FSC parameter (Additional file 1: Fig. S1G). Collectively, these data suggested that loss of Wdr5 results in the blockade of fetal erythropoiesis.
In addition to the defective erythropoiesis observed in the CKO embryos, we wondered whether the impaired hematopoiesis also contributed to the embryonic lethality by loss of Wdr5. We first examined the hematopoietic stem and progenitor cells (HSPCs) in the FLs from CKO or the littermate control embryos at E12.5. The percentage and the absolute cells number of hematopoietic progenitor cells (HPCs) (Lin−Sca1−c-Kit+) or LSK (Lin−Sca1+c-Kit+) were roughly normal in CKO embryos compared with the littermate control at E12.5 (Fig. 2A, B), indicating the initial seeding of HSPCs in the FLs was not influenced in the CKO embryos. We further examined the HSPCs in the FLs from CKO and the littermate control embryos at E13.5 and E15.5 (Fig. 2C, D). We found that the percentage and the absolute cell number of HPCs and LSK cells from FLs were significantly reduced in the CKO embryos compared with the littermate control at E13.5 (Fig. 2E). Intriguingly, the HSPCs underwent rapid expansion in the littermate control FLs but not in the CKO FLs from E13.5-E15.5 (Fig. 2E), indicating the crucial role of Wdr5 in regulating the expansion of HSPCs in FLs. Notably, the expression of c-Kit on Lin− population was slightly decreased in the E12.5 CKO embryos, then further decreased in the E13.5 CKO embryos, and almost diminished in the E15.5 CKO embryos (Fig. 2A, C and D), suggesting Wdr5 might be required for the expression of c-Kit. Surprisingly, the percentage of cKit−Sca1+ population was increased in the CKO FLs at E13.5 and E15.5, so we further dissected this population. Interestingly, a certain percent of CD150+CD48− population was found from c-Kit−Sca1+population in CKO but hardly detected in the littermate control FLs at E15.5 (Fig. 2D, G). By contrast, the CD150+CD48− population was only found in the cKit+Sca1+ population from the control FLs but not the CKO FLs at E13.5 and E15.5, suggesting loss of Wdr5 might alter the HSPCs immunophenotype. Therefore, we performed the colony forming assay to detect the function of HSPCs. Consistently, the CKO FL cells failed to give rise to any colonies in the colony forming assay in vitro (Fig. 2H), indicating loss of Wdr5 impairs thefunction of HSPCs.
In conclusion, we revealed the crucial role of Wdr5 in regulating fetal erythropoiesis as well as hematopoiesis as the de novo findings, which would broaden the understanding of WDR5 function in this field. WDR5 is a very promising therapeutic target in multiple cancers including MLL-rearranged leukemia with genetic model validation and the substantial efforts have been devoted to developing the inhibitors for WDR5 . Our findings for the physiological function of Wdr5 will help to elucidate the safety profile of WDR5 inhibition.
Availability of data and materials
Materials and original data generated in this study are available from the corresponding authors on the reasonable request.
Hematopoietic stem and progenitor cells
Hematopoietic progenitor cells
Guarnaccia AD, Tansey WP. Moonlighting with WDR5: a cellular multitasker. J Clin Med. 2018;7(2):21.
McMahon KA, Hiew SY, Hadjur S, Veiga-Fernandes H, Menzel U, Price AJ, Kioussis D, Williams O, Brady HJ. Mll has a critical role in fetal and adult hematopoietic stem cell self-renewal. Cell Stem Cell. 2007;1(3):338–45.
Jude CD, Climer L, Xu D, Artinger E, Fisher JK, Ernst P. Unique and independent roles for MLL in adult hematopoietic stem cells and progenitors. Cell Stem Cell. 2007;1(3):324–37.
Gan T, Jude CD, Zaffuto K, Ernst P. Developmentally induced Mll1 loss reveals defects in postnatal haematopoiesis. Leukemia. 2010;24(10):1732–41.
Chen C, Liu Y, Rappaport AR, Kitzing T, Schultz N, Zhao Z, Shroff AS, Dickins RA, Vakoc CR, Bradner JE, et al. MLL3 is a haploinsufficient 7q tumor suppressor in acute myeloid leukemia. Cancer Cell. 2014;25(5):652–65.
Santos MA, Faryabi RB, Ergen AV, Day AM, Malhowski A, Canela A, Onozawa M, Lee JE, Callen E, Gutierrez-Martinez P, et al. DNA-damage-induced differentiation of leukaemic cells as an anti-cancer barrier. Nature. 2014;514(7520):107–11.
Tusi BK, Deng C, Salz T, Zeumer L, Li Y, So CW, Morel LM, Qiu Y, Huang S. Setd1a regulates progenitor B-cell-to-precursor B-cell development through histone H3 lysine 4 trimethylation and Ig heavy-chain rearrangement. FASEB J. 2015;29(4):1505–15.
Li Y, Schulz VP, Deng C, Li G, Shen Y, Tusi BK, Ma G, Stees J, Qiu Y, Steiner LA, et al. Setd1a and NURF mediate chromatin dynamics and gene regulation during erythroid lineage commitment and differentiation. Nucleic Acids Res. 2016;44(15):7173–88.
Arndt K, Kranz A, Fohgrub J, Jolly A, Bledau AS, Di Virgilio M, Lesche M, Dahl A, Hofer T, Stewart AF, et al. SETD1A protects HSCs from activation-induced functional decline in vivo. Blood. 2018;131(12):1311–24.
Schmidt K, Zhang Q, Tasdogan A, Petzold A, Dahl A, Arneth BM, Slany R, Fehling HJ, Kranz A, Stewart AF, et al. The H3K4 methyltransferase Setd1b is essential for hematopoietic stem and progenitor cell homeostasis in mice. Elife. 2018. https://doi.org/10.7554/eLife.27157.
Yang Z, Augustin J, Chang C, Hu J, Shah K, Chang CW, Townes T, Jiang H. The DPY30 subunit in SET1/MLL complexes regulates the proliferation and differentiation of hematopoietic progenitor cells. Blood. 2014;124(13):2025–33.
Yang Z, Shah K, Khodadadi-Jamayran A, Jiang H. Dpy30 is critical for maintaining the identity and function of adult hematopoietic stem cells. J Exp Med. 2016;213(11):2349–64.
Liu L, Guo X, Wang Y, Li G, Yu Y, Song Y, Zeng C, Ding Z, Qiu Y, Yan F, Zhang YX, Zhao C, Zhang Y, Dou Y, Atadja P, Li E, Wang H. Loss of Wdr5 attenuates MLL-rearranged leukemogenesis by suppressing Myc targets. Biochim Biophys Acta Mol Basis Dis. 2023;1869(2):166600.
We thank Dr. Jinyong Wang for providing the Vav-iCre mice. We thank Anne Granger, En Li, Rishi Jain and John Tallarico for their support and scientific input, and NIBR Discovery Postdoctoral program for the support of Lulu Liu.
This study was supported by Novartis, the National Key R&D Program of China (2019YFA0111000) and Science and Technology Commission of Shanghai Municipality of China (19JC1414400) to Y.Z.
Ethics approval and consent to participate
All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) in China Novartis Institutes for BioMedical Research Co., Ltd. and the IACUC of Shanghai General Hospital, China.
Consent for publication
All the authors affiliated with Novartis Institutes for BioMedical Research are or were employees of Novartis. The remaining authors declare they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Additional file 1: Table S1.
List of the commercially available antibodies used in this study. Figure S1. Defective erythropoiesis was observed in the CKO embryos, related to Figure 1. (A) Schematic of the genetic targeting strategy to generate Wdr5 KO mice. (B) Representative photographs of the embryos (top) and the FLs (bottom) from CKO and the littermate control at E16.5. (C and D) The absolute cell number of FLs from CKO, heterozygous, and the littermate control embryos at E13.5 (C) and E12.5 (D) (n=2-7 per genotype for each stage). (E) Strategy to analyze fetal erythropoiesis. (F) Representative FACS profile showing the various developmental stages of erythrocytes in the FLs from CKO, heterozygous, and the littermate control embryos at E15.5. (G) Representative FACS profiles to analyze enucleation. Statistical significance is indicated by ns for not statistically significant, *p<0.05, **p<0.01, ***p<0.001, or ****p<0.0001. Data are presented as mean ± SD.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
About this article
Cite this article
Liu, L., Fang, Y., Ding, X. et al. Wdr5 is essential for fetal erythropoiesis and hematopoiesis. Exp Hematol Oncol 12, 39 (2023). https://doi.org/10.1186/s40164-023-00385-3
- Hematopoietic stem cell