American Association for Cancer Research
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Supplementary Materials and Methods, Supplementary Figures 1 through 8 with legends, and Supplementary Tables 1 to 5 from ATF4 Gene Network Mediates Cellular Response to the Anticancer PAD Inhibitor YW3-56 in Triple-Negative Breast Cancer Cells

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posted on 2023-04-03, 14:12 authored by Shu Wang, Xiangyun Amy Chen, Jing Hu, Jian-kang Jiang, Yunfei Li, Ka Yim Chan-Salis, Ying Gu, Gong Chen, Craig Thomas, B. Franklin Pugh, Yanming Wang

The supplementary data contain Supplementary Materials and Methods, Supplementary Figures 1 through 8 with legends, and Supplementary Tables 1 to 5. Supplementary Fig. 1. YW3-56 inhibits growth of p53 wild type and mutant cells. Supplementary Fig. 2. PAD inhibitor YW3-56 inhibits PAD4 in HL-60 derived granulocytic cells. Supplementary Fig. 3. Effects of YW3-56 on the ATF4 and AMPK gene networks. Supplementary Fig. 4. ATF4 directly binds to the SESN2 and DDIT4 promoters. Supplementary Fig. 5. ChIP-exo analyses of ATF4 binding to its target sites. Supplementary Figure 6. mTOR inhibition following thapsigargin induced ER stress. Supplementary Figure 7. Effects of ATF4, SESN2, and DDIT4 depletion on cell growth and YW3-56 mediated cell killing. Supplementary Figure 8. Features of YW3-56 induced cell death. Supplementary Table 1. List of qRT-PCR and ChIP-qPCR primers used in this research work. Supplementary Table 2. Genes with altered expression after YW3-56 treatment. (see Excel spreadsheet) Supplementary Table 3. List of ATF4 associated genes. (see Excel spreadsheet) Supplementary Table 4. List of CEBPB associated genes. (see Excel spreadsheet) Supplementary Table 5. Expression changes of ATF4 and CEBPB candidate genes after YW3-56 treatment.

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ARTICLE ABSTRACT

We previously reported that a pan-PAD inhibitor, YW3-56, activates p53 target genes to inhibit cancer growth. However, the p53-independent anticancer activity and molecular mechanisms of YW3-56 remain largely elusive. Here, gene expression analyses found that ATF4 target genes involved in endoplasmic reticulum (ER) stress response were activated by YW3-56. Depletion of ATF4 greatly attenuated YW3-56–mediated activation of the mTORC1 regulatory genes SESN2 and DDIT4. Using the ChIP-exo method, high-resolution genomic binding sites of ATF4 and CEBPB responsive to YW3-56 treatment were generated. In human breast cancer cells, YW3-56–mediated cell death features mitochondria depletion and autophagy perturbation. Moreover, YW3-56 treatment effectively inhibits the growth of triple-negative breast cancer xenograft tumors in nude mice. Taken together, we unveiled the anticancer mechanisms and therapeutic potentials of the pan-PAD inhibitor YW3-56. Mol Cancer Ther; 14(4); 877–88. ©2015 AACR.