Research ArticleInflammation

The Hippo pathway effector TAZ induces TEAD-dependent liver inflammation and tumors

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Sci. Signal.  11 Sep 2018:
Vol. 11, Issue 547, eaaj1757
DOI: 10.1126/scisignal.aaj1757
  • Fig. 1 TAZ expression correlates with CCL2 and CXCL1 expression in tumors.

    (A and B) Pearson correlation between Taz (A) or Yap (B) expression and chemokine and cytokine mRNAs in RNA-seq data from PDX HCC models. Data for the colored genes are means ± SD from 57 tumor models. (C) Pearson correlation between Yap or Taz expression and Ccl2, Il-6, and Cxcl1 mRNAs in TCGA RNA-seq data sets. Boxplot data are median and quartile correlation values between Taz and Yap with the indicated genes within 22 TCGA indications. (D) Correlation between Yap and Taz expression with inflammatory cytokines. Boxplot data are median and quartile values [left, P value: not significant (ns)], and scatterplot data are mean values (right) from 20 liver cancer cell lines. *P < 0.05 by unpaired t test (B and C), **P < 0.01, ****P < 0.0001 (C), Wilcoxon rank sum test (D, left), or Spearman rank correlation (D, right).

  • Fig. 2 Expression of YAP, not TAZ, in mouse livers increases tumorigenesis.

    (A and B) Survival analysis of FBV-N mice injected with YAP-5SA and YAP-5SA/S94A constructs (A) or YAP-S127A and YAP-S127A/S94A constructs (B). Data are from at least five mice per group analyzed in two independent experiments. (C) Histology analysis of oval cell hyperplasia in liver sections from mice injected with the indicated constructs. Images are representative of five mice per group. Scale bars, 100 μm. (D) Tumor incidence in mice injected with indicated constructs. The y axis indicates number of mice per group. (E and F) Flow cytometry analysis of myeloid cells from the liver of mice injected with the indicated constructs. Dot plots (E) are representative of at least three mice per group. Quantified total white blood cells (WBCs) and myeloid cells in the liver (F) are means ± SEM pooled from all experiments. (G) Survival analysis of FVB-N and C57BL/6 mice injected with TAZ-4SA constructs. Data are from at least three mice per group analyzed in three independent experiments. (H and I) Histology analysis (H) and immunohistochemistry analysis of TAZ abundance (I) in liver sections from mice injected with the indicated constructs. Images are representative of at least three mice per group harvested on day 6 after injection. Scale bars, 100 μm. *P < 0.05 by log rank test (A, B, and G) or by one-sided Barnard’s exact test (D).

  • Fig. 3 Hyperactivated TAZ-4SA mutant cooperates with NRasV12 to induce lethality in mice.

    (A) Survival analysis of mice injected with the indicated TAZ and NRasV12 constructs. Data are from at least four mice per group analyzed in three independent experiments. (B and C) Histology analysis (B) and immunohistochemistry analysis of cytokeratin AE1/AE3 and F4/80 (C) in liver sections from mice injected with indicated constructs. Scale bars, 100 μm. Histology images are representative of at least three mice per group harvested on day 6 after injection. (D and E) Flow cytometry analysis of myeloid cells in the liver of mice injected with the indicated constructs. Dot plots (D) are representative of at least three mice per group from three independent experiments. Quantified total white blood cells (WBCs) and myeloid cells in the liver (E) are means ± SEM pooled from all experiments. (F) Luminex analysis of serum cytokines in mice injected with the indicated constructs. Data are z-scores of means from three to five mice per group, representative of three independent experiments. IFN-γ, interferon-γ; TNF-α, tumor necrosis factor–α; GM-CSF, granulocyte macrophage colony stimulating factor; G-CSF, granulocyte colony stimulating factor; MIP-1α, macrophage inflammatory protein–1α; RANTES, regulated on activation normal T cell expressed and secreted. (G) Fluidigm reverse transcription polymerase chain reaction (RT-PCR) analysis of sorted macrophages from the livers of mice injected with indicated constructs. Data are z-scores of means from three to five mice per group. Rows have been hierarchically clustered using Pearson correlation as a distance metric. *P < 0.05 by log rank test (A) or two-tailed unpaired Student’s t test (E).

  • Fig. 4 Proinflammatory phenotype and lethality induced by TAZ-4SA mutant is dependent on the interaction of TAZ with TEAD.

    (A) Survival analysis of mice injected with indicated constructs. Data are from at least five mice per group analyzed in two independent experiments. (B) Histology analysis of livers from mice injected with indicated constructs. Images are representative of at least three mice per group harvested on day 6 after injection. Scale bars, 100 μm. (C and D) Flow cytometry analysis of myeloid cells in the liver of mice injected with indicated constructs. Dot plots (C) are representative of at least three mice per group from three independent experiments. Quantified total white blood cells (WBCs) and myeloid cells in the liver (D) are means ± SEM pooled from all experiments. (E) Luminex analysis of serum cytokines in mice injected with the indicated constructs. Data are z-scores of means from three to five mice per group. *P < 0.05 by log rank test (A) or two-tailed unpaired Student’s t test (D).

  • Fig. 5 Hyperactivated TAZ mutants cooperate with NRasV12 to induce tumors in mice.

    (A) Survival analysis of mice injected with the indicated 25% TAZ-4SA and NRasV12 constructs. Data are from at least 10 mice per group analyzed in two independent experiments. (B) Histology analysis of liver sections from mice injected with indicated constructs. Images are representative of at least five mice per group. Scale bars, 100 μm. (C) Luminex analysis of serum CCL2 from mice injected with indicated constructs. Data are means ± SEM of at least 10 mice per group pooled from two independent experiments. (D) Survival analysis of mice injected with the indicated TAZ-S89A and NRasV12 constructs. Data are from at least 10 mice per group analyzed in two independent experiments. (E) Tumor incidence in mice injected with indicated constructs. The y axis indicates number of mice per group. (F) Luminex analysis of supernatant cytokines from a cell line derived from mice injected with indicated constructs. Data are means ± SEM of three to five mice per group pooled from two independent experiments. *P < 0.05 by log rank test (A and D), two-tailed unpaired Student’s t test (C and F), or one-side Barnard’s exact test (E).

  • Fig. 6 Differential gene expression profile of YAP- and TAZ-driven liver tumors.

    (A to C) RNA-seq analysis of gene expression in cell lines derived from YAP-S127A, YAP-5SA, and TAZ-S89A mouse liver tumors. Z-score values (A) are from at least three tumors derived from unique mice per group. Fold change in the top 40 most differentially altered genes (B) and normalized expression levels of Cxcl5, Cxcl1, and Ccl2 (C) were determined by DESeq2 analysis. (D and E) qRT-PCR analysis of Cxcl1 (D) and CXCL5 (E) mRNA expression in the indicated cell lines. Data are means ± SEM from three independent experiments. (F) ChIP-qPCR analysis of H3K27ac and total histone H3 in lysates from YAP-S127 and TAZ-S89A tumor-derived cell lines. All YAP and TAZ constructs injected using HTV have a dual Flag and Myc tag. Data are means ± SEM pooled from three independent experiments. *P < 0.05 by two-tailed unpaired Student’s t test (D to F). IgG, immunoglobulin G.

Supplementary Materials

  • www.sciencesignaling.org/cgi/content/full/11/547/eaaj1757/DC1

    Fig. S1. HTV injection of TAZ-4SA + NRasV12 constructs increased liver weights in mice.

    Fig. S2. HTV injection of TAZ constructs increases TAZ abundance in the liver.

    Fig. S3. TAZ alters expression of CD68 and not VWF in mouse liver tissue.

    Fig. S4. Mice injected with TAZ-4SA and NRasV12 had inflammatory cell infiltrates in multiple tissues.

    Fig. S5. Myeloid depletion does not rescue lethality induced by injection with TAZ-4SA + NRasV12.

    Fig. S6. Phenotypic characterization indicates that TAZ regulates liver inflammation in a TEAD-dependent manner.

    Fig. S7. Characterization of liver tumor–derived cell lines from mice injected with expression constructs.

    Fig. S8. Mice injected with 25% TAZ-4SA + NRasV12 constructs developed tumors.

    Fig. S9. YAP and TAZ drive differential gene expression in liver tumor cells.

  • This PDF file includes:

    • Fig. S1. HTV injection of TAZ-4SA + NRasV12 constructs increased liver weights in mice.
    • Fig. S2. HTV injection of TAZ constructs increases TAZ abundance in the liver.
    • Fig. S3. TAZ alters expression of CD68 and not VWF in mouse liver tissue.
    • Fig. S4. Mice injected with TAZ-4SA and NRasV12 had inflammatory cell infiltrates in multiple tissues.
    • Fig. S5. Myeloid depletion does not rescue lethality induced by injection with TAZ-4SA + NRasV12.
    • Fig. S6. Phenotypic characterization indicates that TAZ regulates liver inflammation in a TEAD-dependent manner.
    • Fig. S7. Characterization of liver tumor–derived cell lines from mice injected with expression constructs.
    • Fig. S8. Mice injected with 25% TAZ-4SA + NRasV12 constructs developed tumors.
    • Fig. S9. YAP and TAZ drive differential gene expression in liver tumor cells.

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