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Dysregulation of CITED2 in abnormal lung development in the nitrofen rat model

  • Original Article
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Pediatric Surgery International Aims and scope Submit manuscript

Abstract

Purpose

CITED2 both modulates lung, heart and diaphragm development. The role of CITED2 in the pathogenesis of congenital diaphragmatic hernia (CDH) is unknown. We aimed to study CITED2 during abnormal lung development in the nitrofen model.

Methods

Timed-pregnant rats were given nitrofen on embryonic day (E) 9 to induce CDH. Fetal lungs were harvested on E15, 18 and 21. We performed RT-qPCR, RNAscope™ in situ hybridization and immunofluorescence staining for CITED2.

Results

We observed no difference in RT-qPCR (control: 1.09 ± 0.22 and nitrofen: 0.95 ± 0.18, p = 0.64) and in situ hybridization (1.03 ± 0.03; 1.04 ± 0.03, p = 0.97) for CITED2 expression in E15 nitrofen and control pups. At E18, CITED2 expression was reduced in in situ hybridization of nitrofen lungs (1.47 ± 0.05; 1.14 ± 0.07, p = 0.0006), but not altered in RT-qPCR (1.04 ± 0.16; 0.81 ± 0.13, p = 0.33). In E21 nitrofen lungs, CITED2 RNA expression was increased in RT-qPCR (1.04 ± 0.11; 1.52 ± 0.17, p = 0.03) and in situ hybridization (1.08 ± 0.07, 1.29 ± 0.04, p = 0.02). CITED2 protein abundance was higher in immunofluorescence staining of E21 nitrofen lungs (2.96 × 109 ± 0.13 × 109; 4.82 × 109 ± 0.25 × 109, p < 0.0001).

Conclusion

Our data suggest that dysregulation of CITED2 contributes to abnormal lung development of CDH, as demonstrated by the distinct spatial–temporal distribution in nitrofen-induced lungs.

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Data availability

The data that support the findings of this study are available on request from the corresponding author, RK.

Abbreviations

CDH:

Congenital diaphragmatic hernia

CITED2:

CAMP response element binding protein-binding protein/p300-interacting transactivator with ED (glutamic-asparatic acid)-rich tail

CBP:

CREB-binding protein

References

  1. Zani A, Chung WK, Deprest J et al (2022) Congenital diaphragmatic hernia. Nat Rev Dis Primers 8:1–20. https://doi.org/10.1038/s41572-022-00362-w

    Article  Google Scholar 

  2. Guilbert TW, Gebb SA, Shannon JM (2000) Lung hypoplasia in the nitrofen model of congenital diaphragmatic hernia occurs early in development. Am J Phys-Lung Cellular Molecu Physio 279:L1159–L1171. https://doi.org/10.1152/ajplung.2000.279.6.L1159

    Article  CAS  Google Scholar 

  3. Bamforth SD, Bragança J, Eloranta JJ et al (2001) Cardiac malformations, adrenal agenesis, neural crest defects and exencephaly in mice lacking Cited2, a new Tfap2 co-activator. Nat Genet 29:469–474. https://doi.org/10.1038/ng768

    Article  CAS  PubMed  Google Scholar 

  4. Sun HB, Zhu YX, Yin T et al (1998) MRG1, the product of a melanocyte-specific gene related gene, is a cytokine-inducible transcription factor with transformation activity. Proc Natl Acad Sci 95:13555–13560. https://doi.org/10.1073/pnas.95.23.13555

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Lou X, Sun S, Chen W et al (2011) Negative feedback regulation of NF-κB action by CITED2 in the nucleus. J Immunol 186:539–548. https://doi.org/10.4049/jimmunol.1001650

    Article  CAS  PubMed  Google Scholar 

  6. He Z, Leong DJ, Xu L et al (2019) CITED2 mediates the cross-talk between mechanical loading and IL-4 to promote chondroprotection. Ann N Y Acad Sci 1442:128–137. https://doi.org/10.1111/nyas.14021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Bhattacharya S, Michels CL, Leung M-K et al (1999) Functional role of p35srj, a novel p300/CBP binding protein, during transactivation by HIF-1. Genes Dev 13:64–75. https://doi.org/10.1101/gad.13.1.64

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Xu B, Qu X, Gu S et al (2008) Cited2 is required for fetal lung maturation. Dev Biol 317:95–105. https://doi.org/10.1016/j.ydbio.2008.02.019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Dunwoodie SL, Rodriguez TA, Beddington RSP (1998) Msg1 and Mrg1, founding members of a gene family, show distinct patterns of gene expression during mouse embryogenesis. Mech Dev 72:27–40. https://doi.org/10.1016/S0925-4773(98)00011-2

    Article  CAS  PubMed  Google Scholar 

  10. Weninger WJ, Floro KL, Bennett MB et al (2005) Cited2 is required both for heart morphogenesis and establishment of the left-right axis in mouse development. Development 132:1337–1348. https://doi.org/10.1242/dev.01696

    Article  CAS  PubMed  Google Scholar 

  11. Kuna M, Dhakal P, Iqbal K et al (2023) CITED2 is a conserved regulator of the uterine-placental interface. Proc Natl Acad Sci U S A 120:e2213622120. https://doi.org/10.1073/pnas.2213622120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Yin Z, Haynie J, Yang X et al (2002) The essential role of Cited2, a negative regulator for HIF-1α, in heart development and neurulation. Proc Natl Acad Sci 99:10488–10493. https://doi.org/10.1073/pnas.162371799

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Laresgoiti U, Nikolić MZ, Rao C et al (2016) Lung epithelial tip progenitors integrate glucocorticoid- and STAT3-mediated signals to control progeny fate. Development 143:3686–3699. https://doi.org/10.1242/dev.134023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Li F, Rosenberg E, Smith CI et al (1995) Correlation of expression of transcription factor C/EBP alpha and surfactant protein genes in lung cells. Am J Physiol 269:L241-247. https://doi.org/10.1152/ajplung.1995.269.2.L241

    Article  CAS  PubMed  Google Scholar 

  15. Bassères DS, Levantini E, Ji H et al (2006) Respiratory failure due to differentiation arrest and expansion of alveolar cells following lung-specific loss of the transcription factor C/EBPα in mice. Mol Cell Biol 26:1109–1123. https://doi.org/10.1128/MCB.26.3.1109-1123.2006

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Martis PC, Whitsett JA, Xu Y et al (2006) C/EBPα is required for lung maturation at birth. Development 133:1155–1164. https://doi.org/10.1242/dev.02273

    Article  CAS  PubMed  Google Scholar 

  17. Montalva L, Zani A (2019) Assessment of the nitrofen model of congenital diaphragmatic hernia and of the dysregulated factors involved in pulmonary hypoplasia. Pediatr Surg Int 35:41–61. https://doi.org/10.1007/s00383-018-4375-5

    Article  PubMed  Google Scholar 

  18. Wagner R, Lieckfeldt P, Piyadasa H, et al (2022) Proteomic Profiling of Hypoplastic Lungs Suggests an Underlying Inflammatory Response in the Pathogenesis of Abnormal Lung Development in Congenital Diaphragmatic Hernia. 2022.03.07.483298

  19. Leung M-K, Jones T, Michels CL et al (1999) Molecular Cloning and chromosomal Localization of the human CITED2 gene encoding p35srj/Mrg1. Genomics 61:307–313. https://doi.org/10.1006/geno.1999.5970

    Article  CAS  PubMed  Google Scholar 

  20. CITED2 Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 2 [Homo sapiens (human)] - Gene - NCBI. https://www-ncbi-nlm-nih-gov.ezproxy.medma.uni-heidelberg.de/gene/10370. Accessed 17 Sep 2023

  21. Sperling S, Grimm CH, Dunkel I et al (2005) Identification and functional analysis of CITED2 mutations in patients with congenital heart defects. Hum Mutat 26:575–582. https://doi.org/10.1002/humu.20262

    Article  CAS  PubMed  Google Scholar 

  22. Liu Y, Wang F, Wu Y et al (2014) Variations of CITED2 are associated with congenital heart disease (CHD) in chinese population. PLoS ONE 9:e98157. https://doi.org/10.1371/journal.pone.0098157

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Shen-Schwarz S, Hill LM, Surti U et al (1989) Deletion of terminal portion of 6q: Report of a case with unusual malformations. Am J Med Genet 32:81–86. https://doi.org/10.1002/ajmg.1320320117

    Article  CAS  PubMed  Google Scholar 

  24. Howe DT, Kilby MD, Sirry H et al (1996) Structural chromosome anomalies in congenital diaphragmatic hernia. Prenat Diagn 16:1003–1009. https://doi.org/10.1002/(SICI)1097-0223(199611)16:11%3c1003::AID-PD995%3e3.0.CO;2-D

    Article  CAS  PubMed  Google Scholar 

  25. Tonks A, Wyldes M, Somerset DA et al (2004) Congenital malformations of the diaphragm: findings of the west midlands congenital anomaly register 1995 to 2000. Prenat Diagn 24:596–604. https://doi.org/10.1002/pd.908

    Article  CAS  PubMed  Google Scholar 

  26. Serapiglia V, Stephens CA, Joshi R et al (2022) Fetal tracheal occlusion increases lung basal cells via increased yap signaling. Front Pediatr 9:780166. https://doi.org/10.3389/fped.2021.780166

    Article  PubMed  PubMed Central  Google Scholar 

  27. Boezen HM, Jansen DF, Postma DS (2004) Sex and gender differences in lung development and their clinical significance. Clin Chest Med 25:237–245. https://doi.org/10.1016/j.ccm.2004.01.012

    Article  PubMed  Google Scholar 

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Acknowledgements

MJ is a holder of a scholarship (JA 3507/1-1) from the “Deutsche Forschungsgemeinschaft e.V.”. RK is supported by grants (178347 and 178387) from the Canadian Institutes of Health Research and is the inaugural Thorlakson Chair of Surgical Research for the University of Manitoba.

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Authors and Affiliations

  1. Department of Surgery, Division of Pediatric Surgery, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, and Children’s Hospital Research Institute of Manitoba, AE402-820 Sherbrook Street, Winnipeg, MB, R3A 1S1, Canada

    Marietta Jank, Jacquelyn Schwartz, Yuichiro Miyake, Arzu Ozturk Aptekmann, Daywin Patel & Richard Keijzer

  2. Department of Pediatric Surgery, University Medical Center Mannheim, Heidelberg University, Mannheim, Germany

    Marietta Jank & Michael Boettcher

  3. Department of Pediatric General and Urogenital Surgery, Juntendo University School of Medicine, Tokyo, Japan

    Yuichiro Miyake

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Contributions

MJ, JS and RK: substantial contributions to conception and design, MJ, JS, YM, AOA, DP and RK: acquisition of data, or analysis and interpretation of data, MJ, JS, MB and RK: drafting the article or revising it critically for important intellectual content. All authors: Final approval of the version to be published.

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Correspondence to Richard Keijzer.

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Jank, M., Schwartz, J., Miyake, Y. et al. Dysregulation of CITED2 in abnormal lung development in the nitrofen rat model. Pediatr Surg Int 40, 43 (2024). https://doi.org/10.1007/s00383-023-05607-7

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