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Identification of STXBP6-IRF1 positive feedback loop in regulation of PD-L1 in cancer

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

The clinical success of immune checkpoint blockade against diverse human cancers highlights the critical importance of insightful understanding into mechanisms underlying PD-L1 regulation. IFN-γ released by intratumoral lymphocytes regulates PD-L1 expression in tumor cells through JAK-STAT-IRF1 pathway, while the molecular events prime IRF1 to translocate into nucleus are still obscure. Here we identified STXBP6, previously recognized involving in SNARE complex assembly, negatively regulates PD-L1 transcription via retention of IRF1 in cytoplasm. IFN-γ exposure stimulates accumulation of cytosolic IRF1, which eventually saturates STXBP6 and triggers nuclear translocation of IRF1. Nuclear IRF1 in turn inhibits STXBP6 expression and thereby liberates more IRF1 to migrate to nucleus. Therefore, we identified a novel positive feedback loop between STXBP6 and IRF1 in regulation of PD-L1 expression in cancer. Furthermore, we demonstrate STXBP6 overexpression significantly inhibits T cell activation both in vitro and in vivo. These findings offer new insight into the complexity of PD-L1 expression in cancer and suggest a valuable measure to predict the response to PD-1/PD-L1-based immunotherapy.

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Data availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

ATCC:

American Tissue Culture Collection

ATRA:

All-trans retinoic acid

BSA:

Bovine serum albumin

ChIP:

Chromatin immunoprecipitation

DAB:

Diaminobenzidine

DAPI:

4′,6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s modified Eagle medium

DTT:

Dithiothreitol

ECL:

Enhanced chemiluminescence

ELISA:

Enzyme-linked immunosorbent assay

EMSA:

Electrophoretic mobility shift assay

FBS:

Fetal bovine serum

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

HBSS:

Hank's Balanced Salt Solution

IHC:

Immunohistochemistry

IL-2:

Interleukin-2

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

PFA:

Paraformaldehyde

PVDF:

Polyvinylidene fluoride

RIPA:

Radioimmunoprecipitation assay

TLRs:

Toll-like receptors

SD:

Standard deviation

SLE:

Systemic lupus erythematosus

STR:

Short tandem repeat

SV:

Structural variation

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Funding

This work was supported by National Natural Science Foundation of China, No. 81672796 to Y.B.L. and No. 81171947 to H.H.W.

Author information

Authors and Affiliations

  1. Centre for Translational Medicine, The First Affiliated Hospital, Sun Yat-Sen University, 74 Zhongshan Road II, Guangzhou, 510080, China

    Yanbin Liu, Mingming Zhang & Shulan Yang

  2. Institute of Immunology and Molecular Medicine, Jining Medical University, Jining, 272067, China

    Yanbin Liu

  3. Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-Sen University, 74 Zhongshan Road II, Guangzhou, 510080, China

    Zhicong Huang, Yanli Wei, Mingming Zhang & Haihe Wang

  4. Center for Stem Cell Biology and Tissue Engineering, Key Laboratory of Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China

    Haihe Wang

  5. Longgang District People’s Hospital of Shenzhen, Shenzhen, 5181722, China

    Xingzhi Li

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  1. Yanbin Liu
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  2. Zhicong Huang
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  3. Yanli Wei
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  7. Haihe Wang
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Contributions

Y.B.L. conceived this project and interpreted data. Y.B.L. and H.H.W. wrote the manuscript. Y.B.L., Z.C.H., Y.L.W., M.M.Z. and X.Z.L. performed experiments and analyzed data. Y.B.L., H.H.W. and S.L.Y. supervised the whole study.

Corresponding authors

Correspondence to Yanbin Liu, Shulan Yang or Haihe Wang.

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This study was approved by the Sun Yat-sen University Institutional Review Board.

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Liu, Y., Huang, Z., Wei, Y. et al. Identification of STXBP6-IRF1 positive feedback loop in regulation of PD-L1 in cancer. Cancer Immunol Immunother 70, 275–287 (2021). https://doi.org/10.1007/s00262-020-02678-6

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  • DOI: https://doi.org/10.1007/s00262-020-02678-6

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