In vivo miRNA knockout screening identifies miR-190b as a novel tumor suppressor

PLoS Genet. 2020 Nov 2;16(11):e1009168. doi: 10.1371/journal.pgen.1009168. eCollection 2020 Nov.

Abstract

MicroRNAs (miRNAs) play important roles in the development of various cancers including lung cancer which is one of the devastating diseases worldwide. How miRNAs function in de novo lung tumorigenesis remains largely unknown. We here developed a CRISPR/Cas9-mediated dual guide RNA (dgRNA) system to knockout miRNAs in genetically engineered mouse model (GEMM). Through bioinformatic analyses of human lung cancer miRNA database, we identified 16 downregulated miRNAs associated with malignant progression and performed individual knockout with dgRNA system in KrasG12D/Trp53L/L (KP) mouse model. Using this in vivo knockout screening, we identified miR-30b and miR-146a, which has been previously reported as tumor suppressors and miR-190b, a new tumor-suppressive miRNA in lung cancer development. Over-expression of miR-190b in KP model as well as human lung cancer cell lines significantly suppressed malignant progression. We further found that miR-190b targeted the Hus1 gene and knockout of Hus1 in KP model dramatically suppressed lung tumorigenesis. Collectively, our study developed an in vivo miRNA knockout platform for functionally screening in GEMM and identified miR-190b as a new tumor suppressor in lung cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma of Lung / genetics*
  • Adenocarcinoma of Lung / mortality
  • Adenocarcinoma of Lung / pathology
  • Animals
  • Carcinogenesis / genetics
  • Cell Cycle Proteins / genetics*
  • Cell Line, Tumor
  • Computational Biology
  • Datasets as Topic
  • Disease Models, Animal
  • Down-Regulation
  • Gene Expression Regulation, Neoplastic
  • Genes, Tumor Suppressor
  • HEK293 Cells
  • Humans
  • Kaplan-Meier Estimate
  • Lung / pathology
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / mortality
  • Lung Neoplasms / pathology
  • Mice
  • Mice, Knockout
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Neoplasm Staging
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Tumor Suppressor Protein p53 / genetics

Substances

  • Cell Cycle Proteins
  • HUS1 protein, human
  • Hus1 protein, mouse
  • MIRN190 microRNA, human
  • MIRN190 microRNA, mouse
  • MicroRNAs
  • Trp53 protein, mouse
  • Tumor Suppressor Protein p53
  • Hras protein, mouse
  • Proto-Oncogene Proteins p21(ras)

Grants and funding

This work was supported by the National Basic Research Program of China (2017YFA0505501 to H.J.); Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19020201 to H.J.); the National Natural Science Foundation of China (81430066 to H.J., 91731314 to H.J., 31621003 to H.J., 30971461 to H.J., 81872312 to H.J., 81871875 to L.H., 81802279 to H.H.), the State Key Laboratory of Oncogenes and Related Genes Foundation (KF-20-03 to H.J.), Basic Frontier Scientific Research Program of Chinese Academy of Science (ZDBS-LY-SM006 to H.J.) and the China Postdoctoral Science Foundation (2015M581673 to H.H.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.