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Adenovirus vector-mediated YKL-40 shRNA attenuates eosinophil airway inflammation in a murine asthmatic model

Gene Therapy volume 28, pages 177–185 (2021)Cite this article

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Abstract

Recent studies have revealed that YKL-40 is involved in the pathogenesis of asthma. However, its specific mechanism remains unclear. The present study aims to investigate the effect of adenovirus vector-mediated YKL-40 short hairpin RNA (shRNA) on regulation of airway inflammation in a murine asthmatic model. Mice were assessed for airway hyperresponsiveness (AHR), total leukocytes and the percentage of eosinophil cells in bronchoalveolar lavage fluid (BALF). YKL-40 mRNA and protein expression levels were detected using quantitative real-time PCR and western blot assays. Enzyme-linked immunosorbent assay (ELISA) was used to detect YKL-40 and eosinophil-related chemokine expression levels in BALF and serum. Lung histology analyses were performed to evaluate the degree of inflammatory cell infiltration around the airway and airway mucus secretion.YKL-40 shRNA significantly inhibited the YKL-40 gene expression in asthmatic mice. In addition, YKL-40 shRNA alleviated eosinophilic airway inflammation, AHR, airway mucus secretion and decreased the levels of YKL-40 in BALF and serum in a murine asthmatic model. The levels and mRNA expression of IL-5, IL-13 in asthmatic mice lung tissues, eotaxin, and GM-CSF in BALF and serum significantly decreased. Bone marrow signaling molecules including IL-5, eotaxin, and GM-CSF were correlated with decreased levels of YKL-40. The study reveals that YKL-40 could be involved in asthma inflammation by altering bone marrow signaling molecules. YKL-40 gene RNA interference could provide new therapeutic strategies for asthma.

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Fig. 1: Transfection of adenovirus vector-mediated YKL-40 shRNA down-regulates the expression of YKL-40 in asthmatic mice.
Fig. 2: Transfection of adenovirus vector-mediated YKL-40 shRNA reduced total cell and eosinophil count in BALF.
Fig. 3: Mean percentage increases in lung resistance (RL) with increasing acetylcholine concentration.
Fig. 4: Concentrations of cytokines in BALF and serum analyses by ELISA.
Fig. 5: Suppression of mRNA expression by YKL-40 shRNA.
Fig. 6: Transfection of adenovirus vector-mediated YKL-40 shRNA suppresses infiltration of inflammatory cells and airway mucus secretion.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No: 81570018 and No: 81970022).

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  1. These authors contributed equally: Ling Wang, Aihua Bao, Ying Zheng

Authors and Affiliations

  1. Department of Respiratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Ling Wang, Aihua Bao, Ying Zheng, Aying Ma, Danruo Fang & Suqin Ben

  2. Department of Respiratory Medicine, The Affiliated Hospital of Nantong University, Nantong, China

    Yi Wu

  3. Department of Respiratory Medicine, Chest Hospital of Hebei Province, Shijiazhuang, China

    Huanxia Shang

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Wang, L., Bao, A., Zheng, Y. et al. Adenovirus vector-mediated YKL-40 shRNA attenuates eosinophil airway inflammation in a murine asthmatic model. Gene Ther 28, 177–185 (2021). https://doi.org/10.1038/s41434-020-00202-0

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