Inhibition of O-GlcNAc transferase activates type I interferon-dependent antitumor immunity by bridging cGAS-STING pathway
- Department of Microbial Infection and Immunity, Infectious Disease Institute, The Ohio State University, United States
- Pelotonia Institute for Immuno-Oncology, The Ohio State University Comprehensive Cancer Center, The Ohio State University, United States
- Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, University of Michigan Rogel Cancer Center, University of Michigan, United States
- Department of Cancer Biology and Genetics, The Ohio State University, United States
- Department of Radiation Oncology, The Ohio State University, United States
- Department of Chemistry and Biochemistry, University of Delaware, United States
- Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, United States
Figures
Figure 1 with 1 supplement
OGT is significantly upregulated in human and mouse tumor samples.
(A–C) Boxplot showing mRNA expression level of Ogt, Normal and tumor samples (A), Individual stages (B), Nodal metastasis status (C). The plot was generated using the UALCAN online server. (D–E) …
Figure 1—figure supplement 1
The expression pattern of OGT in TCGA and CTPAC databases.
(A) Boxplot showing mRNA expression level of Ogt in multiple types of cancers. The plot was generated using the GEPIA2 online server. *p<0.05. (B–D) Boxplot showing mRNA expression level of Ogt in …
Figure 2 with 1 supplement
Epithelial OGT deletion inhibits mouse colorectal tumorigenesis.
(A) Western blot analysis of OGT expression in intestinal tissues and counting tumor numbers in APCmin and Ogt IEC cKO mice. (B) Histology analysis of intestinal carcinogenesis by HE staining, scale …
Figure 2—figure supplement 1
The genotype of APCmin,Villin-Cre and Ogtfl/fl mice.
Genomic DNA was extracted from the tails of APCmin, Villin-Cre, and Ogtfl/fl mice and used for PCR with various primers. The resulting products were separated by agarose gel electrophoresis to …
Figure 3 with 1 supplement
OGT deficiency induces cGAS/STING-dependent the type I IFN pathway.
(A–D) Real-time PCR analysis of cytokines mRNA expression in different Ogt−/− cell lines including MC38 (A), LLC (B), HT29 (C), B16-OVA (D) cells. (E) Western blot analysis of the activation of the …
Figure 3—figure supplement 1
In vitro Cross-Priming of T Cells by Ifnar-/- BMDCs.
Ifnar1-/- BMDCs was pre-treated with B16-OVA-Ogt+/+ or B16-OVA-Ogt−/− supernatant, and co-cultured with OT-1 T cell, then T cell proliferation was evaluated by flow cytometry. Representative …
Figure 4 with 1 supplement
OGT deficiency causes DNA damage and accumulates cytosolic DNA.
(A) The extranuclear dsDNA in different Ogt−/− MC38 cells clones were determined by PicoGreen staining assay was quantified by image J. (B) The extranuclear dsDNA in different Ogt−/− MC38 cells …
Figure 4—figure supplement 1
OGT deficiency causes DNA damage and accumulates cytosolic DNA.
(A) The extranuclear dsDNA in different Ogt−/− LLC clones were determined by PicoGreen staining assay and was quantified by image J. (B) The extranuclear dsDNA in different Ogt−/− LLC clones were …
Figure 5 with 1 supplement
The C terminal of HCF-1 rescue DNA damage and the type I IFN pathway in Ogt−/− cells.
(A) Volcano plot of OGT binding proteins identified by LC–MS/MS from stably expressed exogenous GFP-OGT in OGT knockout HT29 cells. (B) OGT and HCF1 binding was confirmed by immunoprecipitation …
Figure 5—figure supplement 1
In vitro pull-down assay analysis of the interaction of HCF-1C600 and OGT.
(A) His pull-down assays were used to analyze the interaction between HCF-1C600 and OGT. (B) His-tagged OGT and HCF1 were expressed and purified, followed by SDS-PAGE separation and staining with …
Figure 6 with 4 supplements
Ogt deficiency inhibits tumor progression through enhancing infiltration by CD8+ T cells.
(A–B) Tumor volume, weight of Ogt+/+ or Ogt−/− MC38 tumors in C57BL/6 J mice and mice survival, n=5 respectively. (C–D) Tumor volume, weight of Ogt+/+ or Ogt−/− LLC tumors in C57BL/6 J mice and mice …
Figure 6—figure supplement 1
The cell proliferation of different tumor model in vitro and B16-OVA tumor growth analysis in vivo.
(A–C), The cell proliferation of different Ogt−/− tumor model in vitro. (A) MC38, (B) LLC, (C) B16-OVA Ogt−/− cells proliferation in vitro. (D–E) Tumor volume, weight of Ogt+/+ or Ogt−/− B16-OVA …
Figure 6—figure supplement 2
Ogt deficiency inhibits tumor progression through enhancing infiltration of CD8+ T cells.
(A) A total schematic of flow cytometry analysis. (B–C) Flow cytometry analysis showing percentage of IFN-γ and TNF-α-expressing intratumoral CD8+ T cells in MC38 tumors with or without PMA and …
Figure 6—figure supplement 3
Ogt deficiency inhibits tumor progression through enhancing infiltration of CD8+ T cells.
(A) Tumor volume and weight of Ogt+/+ or Ogt−/− MC38 tumors injected with control IgG or anti-CD4 antibody at day 0, 7 and 14 post tumor inoculation in C57BL/6 J mice, n=5 respectively. (B–D) The …
Figure 6—figure supplement 4
OGT expression is negatively related to CD8+ T cells infiltration in human colorectal cancer.
(A) Gene Ontology (GO) enrichment and pathway analysis in OGT high and OGT low patients. (B–K) GSEA analysis in OGT high and OGT low patients. T cells activation (B), response to interferon-gamma (C)…
Figure 7 with 1 supplement
Combination therapy with OSMI-1 and anti-PD-L1 augmented T cells and antitumor immunity.
(A) The extranuclear dsDNA were determined by anti-dsDNA fluorescence staining treated with 50 μM and 100 μM OSMI in MC38 cells respectively and was quantified by image J. (B–C) Western blot …
Figure 7—figure supplement 1
OSMI-1 could significantly induce a high percentage of DNA damage.
(A–B) The cell proliferation in different treatments in vitro. (A) MC38 cell proliferation, (B) LLC cell proliferation. (C) The extranuclear dsDNA was measured by anti-dsDNA fluorescence staining …
Additional files
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Supplementary file 1
Primer sequences for genotype, RT-PCR and molecular cloning.
(a) Primer sequences for genotype. (b) Related to Experimental Procedures. Primer sequences for RT-PCR. (c) Related to CRISPR/Cas9. Primer sequences for molecular cloning.
- https://cdn.elifesciences.org/articles/94849/elife-94849-supp1-v1.docx
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Supplementary file 2
Mass spectrometry assay of OGT interactome.
- https://cdn.elifesciences.org/articles/94849/elife-94849-supp2-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/94849/elife-94849-mdarchecklist1-v1.docx
