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Published on: October 19, 2013
Murine Double Hit Model for Neonatal Cardiopulmonary Diseases: Bronchopulmonary Dysplasia (BPD) and Pulmonary
Steven P Garrick1,2, Philip J Berger1,2, Marcel F Nold1,2,3
1Department of Paediatrics, Monash University, Melbourne, Victoria 3168, Australia.
Insights
This study introduces a novel double hit mouse model for bronchopulmonary dysplasia (BPD) and BPD-pulmonary hypertension (BPD-PH), combining antenatal inflammation and postnatal hyperoxia. This model better reflects the multifactorial origins of these complex neonatal lung diseases.
Area of Science:
- Neonatal research
- Pulmonary medicine
- Developmental biology
- Animal models
Background:
- Bronchopulmonary dysplasia (BPD) and associated pulmonary hypertension (BPD-PH) are complex conditions with multifactorial origins.
- Existing murine models often mimic only single risk factors, such as postnatal hyperoxia, failing to fully represent the disease's complexity.
- Preterm infants often require supplemental oxygen (increased fraction of inspired oxygen - FiO2) in neonatal intensive care units, a key risk factor.
Purpose of the Study:
- To develop and validate a more representative murine model for BPD and BPD-PH.
- To combine antenatal systemic inflammation with postnatal hyperoxia to simulate multifactorial disease origins.
- To enable comprehensive multi-omics and imaging analyses from individual animal subjects.
Main Methods:
- A double hit model was established using embryonic day 14 pups.
- Maternal dams received lipopolysaccharide (LPS) to induce antenatal inflammation.
- Postnatally, pups were exposed to either room air (21% FiO2) or hyperoxia (65% FiO2) for up to 28 days.
Main Results:
- The model successfully integrates antenatal inflammation and postnatal hyperoxia.
- Allows for diverse readouts including echocardiography, histology, micro-CT, and immunological analyses (RNA, protein, flow cytometry).
- Provides a platform for studying the combined effects of key BPD and BPD-PH risk factors.
Conclusions:
- This double hit model offers a more comprehensive preclinical tool for investigating BPD and BPD-PH.
- It facilitates a deeper understanding of the interplay between developmental inflammation and oxygen exposure.
- The validated protocol enables robust investigation into therapeutic strategies for these critical neonatal lung conditions.
Abstract:
Bronchopulmonary dysplasia (BPD) and pulmonary hypertension associated with BPD (BPD-PH) are of multifactorial origin and share common risk factors. Most murine models of BPD expose newborn pups to only one of these risk factors-more commonly postnatal hyperoxia-thereby mimicking the vital increased fraction of inspired oxygen (FiO2) that preterm infants in neonatal intensive care units often require. To improve representation of the multifactorial origins of BPD and BPD-PH, we established a double hit model, combining antenatal systemic inflammation followed by postnatal hyperoxia. On embryonic day 14, pups are exposed to systemic maternal inflammation via a single intraperitoneal injection of 150 µg/kg of lipopolysaccharide to the dam. Within 24 h after birth, pups and dams are randomized and exposed to gas with either an FiO2 of 0.21 (room air) or 0.65 (hyperoxia 65%). In our BPD and BPD-PH double hit model, we can obtain multiple readouts from individual pups that include echocardiography, lung histology and immunohistochemistry, ex vivo X-ray micro computed tomography, and pulmonary and plasmatic immunity by RNA, protein, or flow cytometry. This protocol was validated in: Sci Transl Med (2022), DOI: 10.1126/scitranslmed.aaz8454 Graphical abstract Figure 1. Murine double hit model of cardiopulmonary disease. On embryonic day (E)14, pups are exposed to systemic maternal inflammation via a single intraperitoneal injection of 150 µg/kg lipopolysaccharide to the dam. Within 24 h after birth, pups and dams are randomized to be exposed to gas with either a fraction of inspired oxygen (FiO 2 ) of 0.21 (air; 21% O 2 ) or 0.65 (hyperoxia; 65% O 2 ) for a maximum of 28 days. According to the murine stage of lung development ( Schittny, 2017 ), experimental endpoints include postnatal day (D)3, D5, D14, D28, and D60.
