@article{47bfc8565ad54e44b7b2843aaf801015,
title = "Miz1 promotes KRAS-driven lung tumorigenesis by repressing the protocadherin Pcdh10",
abstract = "Targeting KRAS-mutated non-small-cell lung cancer (NSCLC) remains clinically challenging. Here we show that loss of function of Miz1 inhibits lung tumorigenesis in a mouse model of oncogenic KRAS-driven lung cancer. In vitro, knockout or silencing of Miz1 decreases cell proliferation, clonogenicity, migration, invasion, or anchorage-independent growth in mutant (MT) KRAS murine or human NSCLC cells but has unremarkable impact on non-tumorigenic cells or wild-type (WT) KRAS human NSCLC cells. RNA-sequencing reveals Protocadherin-10 (Pcdh10) as the top upregulated gene by Miz1 knockout in MT KRAS murine lung tumor cells. Chromatin immunoprecipitation shows Miz1 binding on the Pcdh10 promoter in MT KRAS lung tumor cells but not non-tumorigenic cells. Importantly, silencing of Pcdh10 rescues cell proliferation and clonogenicity in Miz1 knockout/knockdown MT KRAS murine or human tumor cells, and rescues allograft tumor growth of Miz1 knockout tumor cells in vivo. Miz1 is upregulated in MT KRAS lung tumor tissues compared with adjacent non-involved tissues in mice. Consistent with this, Miz1 is upregulated while Pcdh10 is downregulated in human lung adenocarcinomas (LUAD) compared with normal tissues, and high Miz1 levels or low Pcdh10 levels are associated with poor survival in lung cancer patients. Furthermore, the Miz1 signature is associated with worse survival in MT but not WT KRAS LUAD, and Pcdh10 is downregulated in MT compared to WT KRAS LUAD. Taken together, our studies implicate the Miz1/Pcdh10 axis in oncogenic KRAS-driven lung tumorigenesis.",
keywords = "Gene regulation, KRAS, Miz1, Non-small cell lung cancer, Pcdh10",
author = "Jing Yang and Changchun Hou and Huashan Wang and Perez, {Edith A.} and Do-Umehara, {Hanh Chi} and Huali Dong and Vinothini Arunagiri and Fangjia Tong and {Van Scoyk}, Michelle and Minsu Cho and Xinyi Liu and Xiaodong Ge and Winn, {Robert A.} and Ridge, {Karen M.} and Xiaowei Wang and Chandel, {Navdeep S.} and Jing Liu",
note = "Funding Information: We thank the Cardiovascular Research Core at the University of Illinois at Chicago for the services. We thank Pieter W. Faber and Sandhiya Arun (University of Chicago Genomics Facility) for performing RNA-seq and scRNA-seq. We thank Zhengdeng Lei, Pinal Kanabar, Melline Fontes Noronha, and Mark Maienschein-Cline (Research Informatics Core, Research Resources Center, University of Illinois at Chicago) for the RNA-seq and scRNA-seq analyses. We thank Yale MS & Proteomics Resource for performing the proteomics. JL is supported by the US National Institutes of Health ( HL141459 to J.L.). Funding Information: Dr Liu's work has been funded by the NIH . The authors declare no potential competing interests. Funding Information: Our data showed that Miz1 promotes KRAS-driven lung tumorigenesis. We sought to determine whether Miz1 expression is subject to regulation in the context of oncogenic KRAS-induced lung tumorigenesis. We observed that Miz1 protein was upregulated in tumors from Ad/Cre-treated KRASLSL−G12D/+ mice (Fig. 7A) or KP mice (Fig. 7B) compared to adjacent normal tissues, as analyzed by Miz1 immunohistochemistry (IHC). Quantification of Miz1 staining intensity using HALO (IndicaLab) revealed significantly increased staining in lung tumors compared to adjacent normal tissues from Ad/Cre-treated KP mice (Fig. 7C). Miz1 was also upregulated by MT KRAS in ex vivo AT2 cells isolated from KP mice upon Ad/Cre treatment (Fig. 7D). Miz1 mRNA and protein levels were also increased in a variety of NSCLC cell lines compared with primary normal human bronchial epithelial cells (HBEC) or non-tumorigenic BEAS-2B cells (Fig. 7E and F). Miz1 IHC of lung adenocarcinoma tissue microarray (TMA; Biomax: https://www.biomax.us/tissue-arrays/Lung/LC10013c), which contains 48 cases of lung adenocarcinomas and matched adjacent normal lung tissues, showed that Miz1 was markedly upregulated in lung adenocarcinomas compared to normal tissues (Fig. 7G). Note, A1∼E8 were from 48 cases of lung adenocarcinomas and E9∼J6 were from matched adjacent normal lung tissues, while J7-J10 were from non-related normal lung tissues and J11 was from adrenal gland; and H&E staining of the lung sections were shown side-by-side with Miz1 IHC images. Detailed sample information as well as enlarged H&E staining images of these samples are available at Biomax: https://www.biomax.us/tissue-arrays/Lung/LC10013c. Quantification of Miz1 staining of lung adenocarcinoma TMA using HALO (IndicaLab) revealed significantly increased staining in lung adenocarcinomas compared to matched normal tissues (Fig. 7H). Examples of Miz1 IHC staining of 3 lung adenocarcinomas and matched adjacent normal lung tissues from TMA (Fig. 7G) were shown, in which increased staining for Miz1 in lung tumors compared to the matched adjacent normal tissues was evident (Fig. 7I). To further support this, we also examined NSCLC samples with matched non-involved tissues from the cohort of our University of Illinois at Chicago (UIC) Medical Center. Similar results were obtained from the cohort of our UIC Medical Center (Fig. 7J and K; n = 17). Note: in Fig. 7K, both normal and tumor tissues were shown, which revealed augmented Miz1 expression in tumor tissue compared to adjacent normal tissue. Collectively, our data show that oncogenic KRAS upregulates Miz1, which in turn represses Pcdh10, resulting in increased tumor cell proliferation and promotion of lung tumorigenesis (Fig. 7L).Dr Liu's work has been funded by the NIH. The authors declare no potential competing interests.We thank the Cardiovascular Research Core at the University of Illinois at Chicago for the services. We thank Pieter W. Faber and Sandhiya Arun (University of Chicago Genomics Facility) for performing RNA-seq and scRNA-seq. We thank Zhengdeng Lei, Pinal Kanabar, Melline Fontes Noronha, and Mark Maienschein-Cline (Research Informatics Core, Research Resources Center, University of Illinois at Chicago) for the RNA-seq and scRNA-seq analyses. We thank Yale MS & Proteomics Resource for performing the proteomics. JL is supported by the US National Institutes of Health (HL141459 to J.L.). Publisher Copyright: {\textcopyright} 2022",
year = "2023",
month = feb,
day = "28",
doi = "10.1016/j.canlet.2022.216025",
language = "English (US)",
volume = "555",
journal = "Cancer Letters",
issn = "0304-3835",
publisher = "Elsevier Ireland Ltd",
}
