Zfyve16 regulates the proliferation of B-lymphoid cells

doi:10.1007/s11684-017-0562-3

Front. Med.

2018, Vol. 12

Issue (5) : 559-565 https://doi.org/10.1007/s11684-017-0562-3

RESEARCH ARTICLE

Zfyve16 regulates the proliferation of B-lymphoid cells

Xuemei Zhao¹, Donghe Li¹, Qingsong Qiu¹, Bo Jiao¹, Ruihong Zhang¹, Ping Liu¹(

), Ruibao Ren^1,²(

)

¹. State Key Laboratory for Medical Genomics, Shanghai Institute of Hematology, Collaborative Innovation Center of Hematology, Collaborative Innovation Center of System Biology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
². Department of Biology, Brandeis University, Waltham, MA 02454, USA

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Abstract

Zfyve16 (a.k.a. endofin or endosome-associated FYVE-domain protein), a member of the FYVE-domain protein family, is involved in endosomal trafficking and in TGF-β, BMP, and EGFR signaling. The FYVE protein SARA regulates the TGF-β signaling pathway by recruiting Smad2/3 and accelerating their phosphorylation, thereby altering their susceptibility to TGF-β-mediated T cell suppression. Zfyve16 binds to Smad4 and their binding affects the formation of Smad2/3-Smad4 complex in TGF-β signaling. However, the in vivo function of Zfyve16 remains unknown. In this study, we generated a Zfyve16 knockout mouse strain (Zfyve16^KO) and examined its hematopoietic phenotypes and hematopoietic reconstruction ability. The proportion of T cells in the peripheral blood of Zfyve16^KO mice increases compared with that in wild-type mice. This finding is consistent with the role of Zfyve16 in facilitating TGF-β signaling. Unpredictably, B cell proliferation is inhibited in Zfyve16^KO mice. The proliferation potential of Zfyve16^KO B-lymphoid cells also significantly decreases in vitro. These results suggest that Zfyve16 inhibits the proliferation of T cells, possibly through the TGF-β signaling, but upregulates the proliferation of B-lymphoid cells.

Keywords Zfyve16 endofin hematopoiesis TGF-β lymphocytes

Corresponding Author(s): Ping Liu,Ruibao Ren

Just Accepted Date: 30 October 2017 Online First Date: 18 December 2017 Issue Date: 29 September 2018

Cite this article:

Xuemei Zhao,Donghe Li,Qingsong Qiu, et al. Zfyve16 regulates the proliferation of B-lymphoid cells[J]. Front. Med., 2018, 12(5): 559-565.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-017-0562-3
https://academic.hep.com.cn/fmd/EN/Y2018/V12/I5/559

Fig.1 Targeted disruption of the Zfyve16 gene. (A) Allele architecture and identification of Zfyve16^KO mice. Zfyve16 exons are indicated as gray boxes. FRT and loxP indicate recombinase recognition sites. (B) Zfyve16^KO mice were genotyped by PCR analysis. (C) Representative semi-quantitative reverse-transcription PCR (SqRT-PCR) analysis was performed using RT-F and RT-R primers on exons VI and VII, respectively. RNA was extracted from bone marrow, spleen and liver. (D) Western blot analysis of Zfyve16 protein expression in kidney cells from Zfyve16^KO and wild-type mice. (E) Body weights of Zfyve16^KO and wild-type mice during the 7-month observation period.

Fig.2 Effect of Zfyve16 inactivation on peripheral blood cells. (A) Peripheral white blood cell (WBC), red blood cell (RBC), and platelet (PLT) counts of Zfyve16^KO versus wild-type mice. (B) Percentage of B-lymphoid cells (CD19⁺ B220⁺), T-lymphoid cells (CD3e⁺), and myeloid cells (Gr-1⁺ Mac-1⁺) in the peripheral blood of Zfyve16^KO and wild-type mice analyzed by flow cytometry. (C) Proportion of spleen B cells in each phase of the cell cycle. * P<0.05; ns, no statistical significance.

Fig.3 Effect of Zfyve16 depletion on hematopoietic organs and bone marrow cells. (A) Numbers of bone marrow cells and weights of spleen and liver of Zfyve16^KO and wild-type mice. (B) Percentage of bone marrow hematopoietic stem cells (LSK), myeloid progenitor cells (GMP, CMP, and MEP), and committed lymphoid progenitors (CLP) in Zfyve16^KO and wild-type mice. (C) Percentage of lineage cells in the bone marrow of Zfyve16^KO and wild-type mice. * P<0.05; ns, no statistical significance.

Fig.4 Effect of Zfyve16 on the recovery of hematopoietic cells in response to irradiation. (A) Counts of WBC, RBC, and PLT in Zfyve16^KO and wild-type mice following semi-lethal dose of irradiation. (B) Number of myeloid cells, T-lymphoid cells and B-lymphoid cells in Zfyve16^KO and wild-type mice following semi-lethal dose of irradiation.

Fig.5 Colony-forming ability of bone marrow pre-B cells. (A) Representative photograph of colonies on day 10. (B) Quantitative comparison of colonies formed from the bone marrow of Zfyve16^KO and wild-type mice. *** P<0.001.

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