Skip to main content

Advertisement

Log in

Baseline and innate immune response characterization of a Zfp30 knockout mouse strain

  • Published:
Mammalian Genome Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Data availability

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.

References

  • Charo IF, Ransohoff RM (2006) The many roles of chemokines and chemokine receptors in inflammation. N Engl J Med 354:610–621

    Article  CAS  Google Scholar 

  • Chen W, Schwalie PC, Pankevich EV et al (2019) ZFP30 promotes adipogenesis through the KAP1-mediated activation of a retrotransposon-derived Pparg2 enhancer. Nat Commun 10(1):1809

    Article  Google Scholar 

  • Denny P, Hopes E, Gingles N, Broman K, McPheat W et al (2003) A major locus conferring susceptibility to infection by Streptococcus pneumoniae in mice. Mamm Genome 14:448–453

    Article  Google Scholar 

  • Dickinson M, Flenniken A, Ji X et al (2016) High-throughput discovery of novel developmental phenotypes. Nature 537:508–514

    Article  CAS  Google Scholar 

  • Donoghue LJ, Livraghi-Butrico A, McFadden KM, Thomas JM, Chen G et al (2017) Identification of trans protein QTL for secreted airway mucins in mice and a causal role for Bpifb1. Genetics 207:801–812

    CAS  PubMed  PubMed Central  Google Scholar 

  • Evans CM, Raclawska DS, Ttofali F, Liptzin DR, Fletcher AA et al (2015) The polymeric mucin Muc5ac is required for allergic airway hyperreactivity. Nat Commun 6:6281

    Article  CAS  Google Scholar 

  • Friedman JR, Fredericks WJ, Jensen DE, Speicher DW, Huang XP et al (1996) KAP-1, a novel corepressor for the highly conserved KRAB repression domain. Genes Dev 10:2067–2078

    Article  CAS  Google Scholar 

  • Groner AC, Meylan S, Ciuffi A, Zangger N, Ambrosini G et al (2010) KRAB-zinc finger proteins and KAP1 can mediate long-range transcriptional repression through heterochromatin spreading. PLoS Genet 6(3):e1000869

    Article  Google Scholar 

  • Hornick EE, Banoth B, Miller AM, Zacharias ZR, Jain N et al (2017) Nlrp12 mediates adverse neutrophil recruitment during influenza virus infection. J Immunol 200(3):1188–1197

    Article  Google Scholar 

  • Kelada SNP, Wilson MS, Tavarez U, Kubalanza K, Borate B et al (2011) Strain-dependent genomic factors affect allergen-induced airway hyperresponsiveness in mice. Am J Respir Cell Mol Biol 45:817–824

    Article  CAS  Google Scholar 

  • Keller M, Rabaglia M, Schueler K, Stapleton D, Gatti D, Vincent M et al (2019) Gene loci associated with insulin secretion in islets from nondiabetic mice. J Clin Investig 129(10):4419–4432

    Article  Google Scholar 

  • Kirsten AM, Förster K, Radeczky E, Linnhoff A, Balint B et al (2015) The safety and tolerability of oral AZD5069, a selective CXCR2 antagonist, in patients with moderate-to-severe COPD. Pulm Pharmacol Ther 31:36–41

    Article  CAS  Google Scholar 

  • Laudermilk LT, Thomas JM, Kelada SN (2018) Differential regulation of Zfp30 expression in murine airway epithelia through altered binding of ZFP148 to rs51434084. G3 (Bethesda) 8(2):687–693

    Article  CAS  Google Scholar 

  • Lawlor N, George J, Bolisetty M, Kursawe R, Sun L et al (2017) Single-cell transcriptomes identify human islet cell signatures and reveal cell-type-specific expression changes in type 2 diabetes. Genome Res 27(2):208–222

    Article  CAS  Google Scholar 

  • Li X, Ito M, Zhou F, Youngson N, Zuo X et al (2008) A maternal-zygotic effect of gene Zfp57 maintains both maternal and paternal imprints. Dev Cell 15:547–557

    Article  CAS  Google Scholar 

  • Limjunyawong N, Mock J, Mitzner W (2015) Instillation and fixation methods useful in mouse lung cancer research. J Vis Exp i:1–9

    Google Scholar 

  • Lupo A, Cesaro E, Montano G, Zurlo D, Izzo P et al (2013) KRAB-zinc finger proteins: a repressor family displaying multiple biological functions. Curr Genomics 14:268–278

    Article  CAS  Google Scholar 

  • Matesic LE, De Maio A, Reeves RH (1999) Mapping lipo-polysaccharide response loci in mice using recombinant inbred and congenic strains. Genomics 62:34–41

    Article  CAS  Google Scholar 

  • Medugno L, Florio F, De Cegli R, Grosso M, Lupo A et al (2005) The Krüppel-like zinc-finger protein ZNF224 represses aldolase a gene transcription by interacting with the KAP-1 co-repressor protein. Gene 359:35–43

    Article  CAS  Google Scholar 

  • Mock JR, Tune MK, Dial CF, Torres-Castillo J, Hagan RS, Doerschuk CM (2020) Effects of IFN-γ on immune cell kinetics during the resolution of acute lung injury. Physiol Rep 8(3):e14368

    Article  Google Scholar 

  • Moss RB, Mistry SJ, Konstan MW, Pilewski JM, Kerem E et al (2013) Safety and early treatment effects of the CXCR2 antagonist SB-656933 in patients with cystic fibrosis. J Cyst Fibros 12:241–248

    Article  CAS  Google Scholar 

  • Nair P, Gaga M, Zervas E, Alagha K, Hargreave FE et al (2012) Safety and efficacy of a CXCR2 antagonist in patients with severe asthma and sputum neutrophils: a randomized, placebo-controlled clinical trial. Clin Exp Allergy 42:1097–1103

    Article  CAS  Google Scholar 

  • Nathan C (2006) Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol 6:173–182

    Article  CAS  Google Scholar 

  • Ordoñez CL, Khashayar R, Wong HH, Ferrando R, Wu R et al (2001) Mild and moderate asthma is associated with airway goblet cell hyperplasia and abnormalities in mucin gene expression. Am J Respir Crit Care Med 163:517–523

    Article  Google Scholar 

  • Plasschaert LW, Žillionis R, Choo-Wing R, Savova V, Knehr J et al (2018) A single-cell atlas of the airway epithelium reveals the CFTR-rich pulmonary ionocyte. Nature 560:377–381

    Article  CAS  Google Scholar 

  • Rennard SI, Dale DC, Donohue JF, Kanniess F, Magnussen H et al (2015) CXCR2 antagonist MK-7123 a phase 2 proof-of-concept trial for chronic obstructive pulmonary disease. Am J Respir Crit Care Med 191:1001–1011

    Article  CAS  Google Scholar 

  • Rossi A, Kontarakis Z, Gerri C, Nolte H, Hölper S, Krüger M, Stainier DYR (2015) Genetic compensation induced by deleterious mutations but not gene knockdowns. Nature 524(7564):230–233

    Article  CAS  Google Scholar 

  • Rutledge H, Aylor DL, Carpenter DE, Peck BC, Chines P et al (2014) Genetic regulation of Zfp30, CXCL1, and neutrophilic inflammation in murine lung. Genetics 198:735 LP–745

    Article  Google Scholar 

  • Ryan RF, Schultz DC, Ayyanathan K, Singh PB, Friedman JR et al (1999) KAP-1 corepressor protein interacts and colocalizes with heterochromatic and euchromatic HP1 proteins: a potential role for Krüppel-associated box-zinc finger proteins in heterochromatin-mediated gene silencing. Mol Cell Biol 19:4366–4378

    Article  CAS  Google Scholar 

  • Schultz DC, Ayyanathan K, Negorev D, Maul GG, Rauscher FJ (2002) SETDB1: a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins. Genes Dev 16:919–932

    Article  CAS  Google Scholar 

  • Silver LM (1995) Mouse genetics: concepts and applications. Oxford University Press, New York

    Google Scholar 

  • Sittig LJ, Carbonetto P, Engel KA, Krauss KS, Barrios-Camacho CM, Palmer AA (2016) Genetic background limits generalizability of genotype–phenotype relationships. Neuron 91(6):1253–1259

    Article  CAS  Google Scholar 

  • Smith GJ, Walsh L, Higuchi M, Kelada SNP (2019) Development of a large-scale computer-controlled ozone inhalation exposure system for rodents. Inhal Toxicol 31(2):61–72

    Article  CAS  Google Scholar 

  • Srivastava A, Morgan AP, Najarian ML et al (2017) Genomes of the mouse collaborative cross. Genetics 206(2):537–556

    Article  CAS  Google Scholar 

  • Takahashi K, Sugi Y, Hosono A, Kaminogawa S (2009) Epigenetic regulation of TLR4 gene expression in intestinal epithelial cells for the maintenance of intestinal homeostasis. J Immunol 183:6522–6529

    Article  CAS  Google Scholar 

  • Todd CM, Salter BM, Murphy DM, Watson RM, Howie KJ et al (2016) The effects of a CXCR1/CXCR2 antagonist on neutrophil migration in mild atopic asthmatic subjects. Pulm Pharmacol Ther 41:34–39

    Article  CAS  Google Scholar 

  • Ulland TK, Jain N, Hornick EE, Elliott EI, Clay GM et al (2016) Nlrp12 mutation causes C57BL/6J strain-specific defect in neutrophil recruitment. Nat Commun 7:1–13

    Article  Google Scholar 

  • Urrutia R (2003) KRAB-containing zinc-finger repressor proteins. Genome Biol 4:231.1–231.8

    Article  Google Scholar 

  • Watz H, Uddin M, Pedersen F, Kirsten A, Goldmann T et al (2017) Effects of the CXCR2 antagonist AZD5069 on lung neutrophil recruitment in asthma. Pulm Pharmacol Ther 45:121–123

    Article  CAS  Google Scholar 

  • You Y, Brody S (2013) Culture and differentiation of mouse tracheal epithelial cells. Second Edition, Epithelial Cell Culture Protocols, pp 123–143

    Google Scholar 

Download references

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.

Author information

Authors and Affiliations

Authors

Contributions

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.

Corresponding author

Correspondence to Samir N. P. Kelada.

Ethics declarations

Conflict of interest

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.

Ethical approval

Not applicable.

Informed consent

Not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 6874 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00335-020-09847-z

Navigation