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Effects of systemic anticoagulation in a murine model of compensatory lung growth

Pediatric Research volume 93, pages 1846–1855 (2023)Cite this article

Abstract

Background

Neonates with congenital diaphragmatic hernia (CDH) suffer from pulmonary hypoplasia (PH) and may require extracorporeal membrane oxygenation (ECMO) and anticoagulation, often with unfractionated heparin (UFH). UFH interacts with vascular endothelial growth factor (VEGF), a factor important in lung development. We investigated the effects of UFH, low molecular weight heparin (LMWH), and bivalirudin (BV) on a murine model of compensatory lung growth (CLG).

Methods

Proliferation and apoptosis were assessed in microvascular lung endothelial cells (HMVEC-L) treated with anticoagulants. Eight-week-old C57Bl/6J mice underwent left pneumonectomy and anticoagulation with low- or high-dose UFH, LMWH, BV, or saline control. Lung volume, pulmonary function tests, morphometrics, treadmill exercise tolerance, and pulmonary protein expression were examined.

Results

UFH and LMWH inhibited HMVEC-L proliferation. BV promoted proliferation and decreased apoptosis. UFH and LMWH-treated mice had reduced lung volume, total lung capacity, alveolar volume, and septal surface area compared to controls, while BV did not affect these measures. UFH and LMWH-treated mice had lower exercise tolerance compared to controls.

Conclusions

UFH and LMWH impair pulmonary growth, alveolarization, and exercise tolerance, while BV does not. Alternative anticoagulants to heparin may be considered to improve functional outcomes for neonates with CDH and pulmonary hypoplasia.

Impact

  • Unfractionated heparin and low molecular weight heparin may modify compensatory lung growth by reducing microvascular lung endothelial cell proliferation and affecting pulmonary angiogenic signaling.

  • Functional effects of unfractionated heparin and low molecular weight heparin on murine compensatory lung growth include reduction in exercise tolerance.

  • Bivalirudin, a direct thrombin inhibitor, may increase microvascular lung endothelial cell proliferation and preserves lung volume, alveolarization, and exercise tolerance in a murine compensatory lung growth model.

  • Anticoagulants alternative to heparin should be further investigated for use in neonates with pulmonary hypoplastic diseases to optimize lung growth and development and improve outcomes.

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Fig. 1: Unfractionated and low molecular weight heparin inhibit endothelial cell proliferation and signaling.
Fig. 2: UFH and LMWH inhibit compensatory lung growth (CLG) and subsequent pulmonary function.
Fig. 3: UFH and LMWH impair alveolarization in CLG.
Fig. 4: UFH and LMWH impair exercise tolerance after left pneumonectomy.
Fig. 5: Anticoagulation with UFH or LMWH affect angiogenic and proliferative signaling.
Fig. 6: Bivalirudin (BV) has minimal detrimental effects on endothelial cell proliferation or CLG.

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Data availability

All data generated or analyzed during this study are included in this published article (and its Supplementary Information files). For further details regarding the methods or materials, please contact the corresponding author.

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Acknowledgements

The authors acknowledge the neurodevelopmental behavior core at Boston Children’s Hospital CHB IDDRC, 1U54HD090255.

Funding

Research funding for this study was provided by the Richard and Sandra Cummings Research Fellowship of the Beth Israel Deaconess Medical Center Department of Surgery (L.J.Y.), Boston Children’s Hospital Surgical Foundation (L.J.Y., V.H.K., J.D.S., M.P.), The Boston Children’s Hospital Vascular Biology Program (all), the Joshua Ryan Rappaport Fellowship of the Boston Children’s Hospital Department of Surgery (L.J.Y.), the National Institutes of Health grants 2T32DK007754–21 (S.T.T.) and 5T32HL007734 (D.T.D., S.C.F.), and the Howard Hughes Medical Institute (B.S.C.).

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Author notes

  1. These authors contributed equally: Lumeng J. Yu, Victoria H. Ko.

Authors and Affiliations

  1. Vascular Biology Program, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA

    Lumeng J. Yu, Victoria H. Ko, Savas T. Tsikis, Duy T. Dao, Jordan D. Secor, Amy Pan, Bennet S. Cho, Scott C. Fligor, Hiroko Kishikawa & Mark Puder

  2. Department of Surgery, Boston Children’s Hospital, Harvard Medical School, 300 Longwood Avenue, Fegan 3, Boston, MA, USA

    Lumeng J. Yu, Victoria H. Ko, Savas T. Tsikis, Duy T. Dao, Jordan D. Secor, Amy Pan, Bennet S. Cho, Scott C. Fligor, Hiroko Kishikawa & Mark Puder

  3. Institutional Centers for Clinical and Translational Research, Boston Children’s Hospital, Boston, MA, USA

    Paul D. Michell

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Contributions

Conceptualization: V.H.K., L.J.Y., D.T.D., M.P. Data curation: L.J.Y., V.H.K., S.T.T., J.D.S., A.P., B.S.C., S.C.F. Formal analysis: L.J.Y., V.H.K., P.D.M. Funding acquisition: L.J.Y., M.P. Investigation: V.H.K., L.J.Y., S.T.T., D.T.D., A.P. Methodology: V.H.K., L.J.Y., S.T.T., D.T.D., S.C.F. Project administration: A.P. Supervision: M.P. Writing—original draft: L.J.Y., V.H.K., S.T.T. Writing—review and editing: L.J.Y., S.T.T., V.H.K., D.T.D., J.D.S., P.D.M., S.C.F., B.S.C., H.K., M.P.

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Correspondence to Mark Puder.

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Yu, L.J., Ko, V.H., Tsikis, S.T. et al. Effects of systemic anticoagulation in a murine model of compensatory lung growth. Pediatr Res 93, 1846–1855 (2023). https://doi.org/10.1038/s41390-022-02323-1

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