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. 2022 Sep;24(9):1785-1799.
doi: 10.1007/s12094-022-02835-6. Epub 2022 Apr 29.

Integrated analysis of ALK higher expression in human cancer and downregulation in LUAD using RNA molecular scissors

Saifullah  1   2 , Toshifumi Tsukahara  3   4
Affiliations

Integrated analysis of ALK higher expression in human cancer and downregulation in LUAD using RNA molecular scissors

Saifullah et al. Clin Transl Oncol. 2022 Sep.

Abstract

Purpose: Anaplastic lymphoma kinase (ALK) is an endorsed molecular target in ALK-rearranged carcinomas, including lung adenocarcinoma. However, the clinical advantage of targeting ALK using druggable inhibitors is almost universally restricted by the development of drug resistance. Therefore, a strategy for combating ALK overexpression remains paramount for ALK-driven cancer.

Methods: We systemically analyzed the overexpression pattern of ALK and its clinical consequences, genetic alterations, and their significance in cancer hallmark genes, and correlation using integrated multidimensional approaches. The LwCas13a RNA molecular scissors was used to downregulate ALK-rearrangement by leveraging two target guide RNAs in lung adenocarcinoma (LUAD) cells. Immunocytochemistry, immunoblotting, and MTT assays were conducted to validate the downregulation.

Results: We found elevated levels of ALK in several malignancies, including LUAD, than in normal tissues. Higher expression of ALK was significantly associated with worse or shorter survival than patients with lower expression. We identified numerous genetic alterations in ALK, which potentially alter the cancer hallmark genes, including STAT1 and CTSL, in patients with LUAD. Next, we observed that the LwCas13a molecular scissors robustly downregulated both phosphorylated and total ALK chimera protein expression in LUAD cells compared to the control. Furthermore, we found that downregulation of ALK chimera protein substantially inhibited cell viability and induced cell death, including apoptosis.

Conclusion: Our findings suggest a basis for ALK as a prognostic biomarker and the LwCas13a molecular scissors successfully downregulated the onco-driver ALK-rearrangement protein, which will potentially pave the way toward the development of novel therapeutic strategies for ALK-driven cancer.

Keywords: ALK mutation; ALK overexpression; LUAD; LwCas13a; Molecular therapy.

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References

    1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71:209–49. https://doi.org/10.3322/caac.21660 . - DOI - PubMed
    1. Carrera S, Buque A, Azkona E, Aresti U, Calvo B, Sancho A, Arruti M, Nuño M, Rubio I, de Lobera AR, et al. Epidermal growth factor receptor tyrosine-kinase inhibitor treatment resistance in non-small cell lung cancer: biological basis and therapeutic strategies. Clin Transl Oncol. 2014;16:339–50. https://doi.org/10.1007/s12094-013-1143-9 . - DOI - PubMed
    1. Soda M, Choi YL, Enomoto M, Takada S, Yamashita Y, Ishikawa S, Fujiwara S-I, Watanabe H, Kurashina K, Hatanaka H, et al. Identification of the transforming EML4–ALK fusion gene in non-small-cell lung cancer. Nature. 2007;448:561–6. https://doi.org/10.1038/nature05945 . - DOI - PubMed
    1. Sabir SR, Yeoh S, Jackson G, Bayliss R. EML4-ALK variants: biological and molecular properties, and the implications for patients. Cancers. 2017;9:118. - DOI
    1. Paez JG, Jänne PA, Lee JC, Tracy S, Greulich H, Gabriel S, Herman P, Kaye FJ, Lindeman N, Boggon TJ et al. EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science (New York, N.Y.) 2004; 304, 1497–1500. https://doi.org/10.1126/science.1099314.