Abstract
Airway neutrophilia is correlated with disease severity in a number of chronic and acute pulmonary diseases, and dysregulation of neutrophil chemotaxis can lead to host tissue damage. The gene Zfp30 was previously identified as a candidate regulator of neutrophil recruitment to the lungs and secretion of CXCL1, a potent neutrophil chemokine, in a genome-wide mapping study using the Collaborative Cross. ZFP30 is a putative transcriptional repressor with a KRAB domain capable of inducing heterochromatin formation. Using a CRISPR-mediated knockout mouse model, we investigated the role that Zfp30 plays in recruitment of neutrophils to the lung using models of allergic airway disease and acute lung injury. We found that the Zfp30 null allele did not affect CXCL1 secretion or neutrophil recruitment to the lungs in response to various innate immune stimuli. Intriguingly, despite the lack of neutrophil phenotype, we found there was a significant reduction in the proportion of live Zfp30 homozygous female mutant mice produced from heterozygous matings. This deviation from the expected Mendelian ratios implicates Zfp30 in fertility or embryonic development. Overall, our results indicate that Zfp30 is an essential gene but does not influence neutrophilic inflammation in this particular knockout model.
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The datasets generated during and/or analyzed during the current study will be made available as a data supplement if/when the manuscript is accepted.
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Acknowledgements
The authors would like to thank Gregory J. Smith, Ph.D. for his assistance with in vivo ozone exposures; Larry Ostrowski, Ph.D. and Ximena Bustamante, Ph.D. for their assistance with MTEC isolation and culture; Kim Burns for her assistance with histology; Max Lowman for technical assistance with qPCR work; Autumn Sanson for her assistance with in vivo data generation; Gang Chen for his suggestions regarding mouse tracheal epithelial cell qPCR; Praveen Sethupathy, Ph.D. and Yu-Han Hung, Ph.D. for their consultation on metabolic phenotypes; and David Aylor, Ph.D. for input on statistical analysis of genotype ratios. The authors would additionally like to thank the UNC CGIBD Advanced Analytics Core for their work on cytokine multiplex assays, the Animal Histopathology and Laboratory Medicine Core for their work in processing complete blood count assays, and the UNC NORC Animal Metabolism Phenotyping core for their work on mouse MRIs.
Funding
This work was supported by NIH Grants ES024965 and HL122711. The UNC NORC Animal Metabolism Phenotyping Core is supported by DK056350. The UNC CGIBD Advanced Analytics core is supported by DK034987. The UNC Animal Histopathology Core is supported in part by an NCI Center Core Support Grant (5P30CA016086-41) to the UNC Lineberger Comprehensive Cancer Center.
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by LTL, AT, AKH, JMT, KMM, MKT, DOC, JRM, and SNPK.
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Dale Cowley is employed by, has Equity Ownership in, and serves on the Board of Directors of TransViragen, the company which has been contracted by UNC-Chapel Hill to manage its Animal Models Core Facility.
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Laudermilk, L.T., Tovar, A., Homstad, A.K. et al. Baseline and innate immune response characterization of a Zfp30 knockout mouse strain. Mamm Genome 31, 205–214 (2020). https://doi.org/10.1007/s00335-020-09847-z
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DOI: https://doi.org/10.1007/s00335-020-09847-z