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Genome-wide CRISPR-Cas9 screening identifies ITGA8 responsible for abivertinib sensitivity in lung adenocarcinoma

Acta Pharmacologica Sinica volume 46, pages 1419–1432 (2025)Cite this article

Abstract

The emergence of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) has improved the prognosis for lung cancer patients with EGFR-driven mutations. However, acquired resistance to EGFR-TKIs poses a significant challenge to the treatment. Overcoming the resistance has primarily focused on developing next-generation targeted therapies based on the molecular mechanisms of resistance or inhibiting the activation of bypass pathways to suppress or reverse the resistance. In this study we developed a novel approach by using CRISPR-Cas9 whole-genome library screening to identify the genes that enhance the sensitivity of lung adenocarcinoma cells to EGFR-TKIs. Through this screening, we revealed integrin subunit alpha 8 (ITGA8) as the key gene that enhanced sensitivity to abivertinib in lung adenocarcinoma. Notably, ITGA8 expression was significantly downregulated in lung adenocarcinoma tissues compared to adjacent normal tissues. Bioinformatics analyses revealed that ITGA8 was positively correlated with the sensitivity of lung adenocarcinoma to abivertinib. We showed that knockdown of ITGA8 significantly enhanced the proliferation, migration and invasion of H1975 cells. Conversely, overexpression of ITGA8 reduced the proliferation migration and invasion of H1975/ABIR cells. Furthermore, we demonstrated that ITGA8 sensitized lung adenocarcinoma cells to EGFR-TKIs by attenuating the downstream FAK/SRC/AKT/MAPK signaling pathway. In H1975 cell xenograft mouse models, knockdown of ITGA8 significantly increased tumor growth and reduced the sensitivity to abivertinib, whereas overexpression of ITGA8 markedly suppressed tumor proliferation and enhanced sensitivity to the drug. This study demonstrates that ITGA8 inhibits the proliferation, invasion and migration of lung adenocarcinoma cells, enhances the sensitivity to EGFR-TKIs, improves treatment efficacy, and delays the progression of acquired resistance. Thus, ITGA8 presents a potential therapeutic candidate for addressing acquired resistance to EGFR-TKIs from a novel perspective.

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Fig. 1: Identifying genes enhancing sensitivity to abivertinib in lung adenocarcinoma through CRISPR-Cas9 genome-wide library screening.
Fig. 2: Expression differences of ITGA8 in tumor tissues and adjacent tissues.
Fig. 3: ITGA8 enhanced sensitivity of lung adenocarcinoma to abivertinib and inhibited migration, invasion, and proliferation capacities of lung adenocarcinoma.
Fig. 4: Impact of ITGA8 expression levels on proliferation, migration, invasion capacities, and drug resistance of H1975/AR.
Fig. 5: Effects of abivertinib and ITGA8 on the EGFR/FAK/SRC/AKT/MAPK pathway.
Fig. 6: Changes in FAK expression levels rescued the downstream pathway alterations, as well as changes in migration, invasion, proliferation capacities, and drug sensitivity induced by ITGA8 regulation.
Fig. 7: ITGA8 specifically binds to downstream FAK.
Fig. 8: Xenograft tumor model in nude mice.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (grants 82172569, 82072595), the Natural Science Foundation of Tianjin (23JCZDJC00710, 23JCYBJC01010), the Tianjin Key Medical Discipline (Specialty) Construction Project (TJYXZDXK-061B), the Tianjin Health Science and Technology Project (TJWJ2022XK005), and Science Foundation of Tianjin Educational Commission Fund (2020KJ157, 2020KJ155).

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  1. These authors contributed equally: Xuan-guang Li, Guang-sheng Zhu, Pei-jun Cao

Authors and Affiliations

  1. Department of Lung Cancer Surgery, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Xuan-guang Li, Guang-sheng Zhu, Pei-jun Cao, Hua Huang, Yu-hao Chen, Pei-jie Chen, Di Wu, Zi-he Zhang, Rui-hao Zhang, Zi-xuan Hu, Wen-hao Zhao, Ming-hui Liu & Jun Chen

  2. Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Xuan-guang Li, Chen Chen, Chen Ding, Yong-wen Li, Hong-yu Liu & Jun Chen

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  1. Xuan-guang Li
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Contributions

JC, HYL, and YWL contributed to the study conception and design. XGL, GSZ, and PJC conducted the data analysis. HH, YHC, and CC performed the data collection. PJC, DW, CD, ZHZ, RHZ, ZXH, WHZ, and MHL conducted material preparation. The first draft of the manuscript was written by XGL and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Yong-wen Li, Hong-yu Liu or Jun Chen.

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Li, Xg., Zhu, Gs., Cao, Pj. et al. Genome-wide CRISPR-Cas9 screening identifies ITGA8 responsible for abivertinib sensitivity in lung adenocarcinoma. Acta Pharmacol Sin 46, 1419–1432 (2025). https://doi.org/10.1038/s41401-024-01451-0

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