<p>This file contains the following: Supplementary Figure 1: Representative H&E staining and immunohistochemistry for keratin 5 of a metastasis in the lung of a tumor-bearing TAp63-/- mouse. Supplementary Figure 2: TAp63-deficient tumors exhibit deregulated mRNA and miRNA expression. Supplementary Figure 3: TAp63-regulated miR-30c-2* and miR-497-5p suppresses cuSCC through induction of cell death and cell cycle arrest. Supplementary Figure 4: Identification of miRNA targets through the use of TMT-LC-MS/MS. Supplementary Figure 5: Inhibition of miR-30c-2-3p and miR-497-5p targets affects cuSCC cell proliferation and survival. Supplementary Figure 6: AURKA is a viable therapeutic target in cuSCC. Supplementary Methods and Materials. Supplementary References.</p>
<p>Supplementary Table 1: Differentially expressed genes in TAp63-/- cuSCC vs. TAp63-/- skin Supplementary Table 2: Differentially expressed miRNAs in TAp63-/- cuSCC vs. TAp63-/- skin Supplementary Table 3: microRNA-mRNA pair analysis of TAp63-/- cuSCC vs. TAp63-/- skin: Underexpressed miRNAs and overexpressed mRNAs. Supplementary Table 4: microRNA-mRNA pair analysis of TAp63-/- cuSCC vs. TAp63-/- skin: Overexpressed miRNAs and underexpressed mRNAs. Supplementary Table 5: Differentially expressed mRNAs from TAp63-/- cuSCC vs cuSCC and Human cuSCC vs. skin. Supplementary Table 6: Differentially expressed miRNAs from TAp63-/- cuSCC vs cuSCC and Human cuSCC vs. skin. Supplementary Table 7: GSEA of common differentially expressed mRNAs in TAp63-/- cuSCC and human cuSCC. Supplementary Table 8: Differentially expressed proteins in miR-30c-2* mimic transfected COLO16 cells measured by TMT-6-plex LC-MS/MS. Supplementary Table 9: Differentially expressed proteins in miR-497 mimic transfected COLO16 cells measured by TMT-6-plex LC-MS/MS. Supplementary Table 10: miRNA target prediction of corresponding mRNAs for the differentially expressed proteins in the miR-30c-2* transfected COLO16 cells. Supplementary Table 11: miRNA target prediction of corresponding mRNAs for the differentially expressed proteins in the miR-497 transfected COLO16 cells. Supplementary Table 12: Overexpressed mRNAs in both mouse TAp63-/- cuSCC and human cuSCC RNA-Seq signatures</p>
<div>Abstract<p>TAp63 is a p53 family member and potent tumor and metastasis suppressor. Here, we show that <i>TAp63<sup>−/−</sup></i> mice exhibit an increased susceptibility to ultraviolet radiation–induced cutaneous squamous cell carcinoma (cuSCC). A human-to-mouse comparison of cuSCC tumors identified miR-30c-2* and miR-497 as underexpressed in TAp63-deficient cuSCC. Reintroduction of these miRNAs significantly inhibited the growth of cuSCC cell lines and tumors. Proteomic profiling of cells expressing either miRNA showed downregulation of cell-cycle progression and mitosis-associated proteins. A mouse to human and cross-platform comparison of RNA-sequencing and proteomics data identified a 7-gene signature, including <i>AURKA, KIF18B, PKMYT1</i>, and <i>ORC1</i>, which were overexpressed in cuSCC. Knockdown of these factors in cuSCC cell lines suppressed tumor cell proliferation and induced apoptosis. In addition, selective inhibition of AURKA suppressed cuSCC cell proliferation, induced apoptosis, and showed antitumor effects <i>in vivo</i>. Finally, treatment with miR-30c-2* or miR-497 miRNA mimics was highly effective in suppressing cuSCC growth <i>in vivo</i>. Our data establish TAp63 as an essential regulator of novel miRNAs that can be therapeutically targeted for potent suppression of cuSCC.</p>Significance:<p>This study provides preclinical evidence for the use of miR-30c-2*/miR-497 delivery and AURKA inhibition in the treatment of cuSCC, which currently has no FDA-approved targeted therapies.</p><p><i>See related commentary by Parrales and Iwakuma, p. 2439</i></p></div>
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.