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Aberrant PGC-1α signaling in a lamb model of persistent pulmonary hypertension of the newborn

Pediatric Research (2024)Cite this article

Abstract

Background

Persistent Pulmonary Hypertension of the Newborn (PPHN) is characterized by elevated pulmonary vascular resistance (PVR), resulting in hypoxemia. Impaired angiogenesis contributes to high PVR. Pulmonary artery endothelial cells (PAECs) in PPHN exhibit decreased mitochondrial respiration and angiogenesis. We hypothesize that Peroxisome Proliferator-Activated Receptor Gamma Co-Activator-1α (PGC-1α) downregulation leads to reduced mitochondrial function and angiogenesis in PPHN.

Methods

Studies were performed in PAECs isolated from fetal lambs with PPHN induced by ductus arteriosus constriction, with gestation-matched controls and in normal human umbilical vein endothelial cells (HUVECs). PGC-1α was knocked downed in control lamb PAECs and HUVECs and overexpressed in PPHN PAECs to investigate the effects on mitochondrial function and angiogenesis.

Results

PPHN PAECs had decreased PGC-1α expression compared to controls. PGC-1α knockdown in HUVECs led to reduced Nuclear Respiratory Factor-1 (NRF-1), Transcription Factor-A of Mitochondria (TFAM), and mitochondrial electron transport chain (ETC) complexes expression. PGC-1α knockdown in control PAECs led to decreased in vitro capillary tube formation, cell migration, and proliferation. PGC-1α upregulation in PPHN PAECs led to increased ETC complexes expression and improved tube formation, cell migration, and proliferation.

Conclusion

PGC-1α downregulation contributes to reduced mitochondrial oxidative phosphorylation through control of the ETC complexes, thereby affecting angiogenesis in PPHN.

Impact

  • Reveals a novel mechanism for angiogenesis dysfunction in persistent pulmonary hypertension of the newborn (PPHN).

  • Identifies a key mitochondrial transcription factor, Peroxisome Proliferator-Activated Receptor Gamma Co-Activator-1α (PGC-1α), as contributing to the altered adaptation and impaired angiogenesis function that characterizes PPHN through its regulation of mitochondrial function and oxidative phosphorylation.

  • May provide translational significance as this mechanism offers a new therapeutic target in PPHN, and efforts to restore PGC-1α expression may improve postnatal transition in PPHN.

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Fig. 1: Immunoblotting for PGC-1α protein levels in lamb PAECs.
Fig. 2: Immunoblotting for mitochondrial transcription factors and mitochondrial electron transport chain (ETC) complex proteins following PGC-1α knockdown (KD).
Fig. 3: Immunoblotting for mitochondrial electron transport chain (ETC) complex proteins following PGC-1α overexpression (OE).
Fig. 4: Tube formation assay comparing control and PPHN PAECs with genetically-modified PGC-1α expression.
Fig. 5: Angiogenesis assays comparing control and PPHN PAECs with genetically-modified PGC-1α expression.
Fig. 6: Assessing PGC-1α’s role in the regulation of mitochondrial biogenesis and redox balance in lamb PAECs.

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Funding

This work of G.G.K. was supported by 1R01HL 136597-01 grant from U.S. National Heart, Lung, & Blood Institute (NHLBI); Children’s Research Institute Pilot Innovation Research Award for Muma Endowed Chair in Neonatology; and Advancing a Healthier Wisconsin Foundation Endowment.

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Authors and Affiliations

  1. Institutional Affiliation (of all authors): Department of Pediatrics, Division of Neonatology, Medical College of Wisconsin (MCW), Milwaukee, WI, USA

    Emily A. Mooers, Hollis M. Johnson, Teresa Michalkiewicz, Ujala Rana, Chintamani Joshi, Adeleye J. Afolayan, Ru-Jeng Teng & Girija G. Konduri

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  1. Emily A. Mooers
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  2. Hollis M. Johnson
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Contributions

E.A.M.—had substantial contributions to conception and design; acquisition of data; analysis and interpretation of data; drafting the article and revising article critically for important intellectual content; and approving the version to be published. H.M.J.—had substantial contributions to acquisition of data. T.M.—had substantial contributions to acquisition of data as well as analysis and interpretation of data. U.R.—had substantial contributions to acquisition of data. C.J.—had substantial contributions to acquisition of data as well as analysis and interpretation of data. A.J.A.—had substantial contributions to conception and design as well as analysis and interpretation of data. R.-J.T.—had substantial contributions to conception and design as well as analysis and interpretation of data. G.G.K.—had substantial contributions to conception and design; analysis and interpretation of data; revising articles critically for important intellectual content; and approving the final version to be published.

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Correspondence to Emily A. Mooers.

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Mooers, E.A., Johnson, H.M., Michalkiewicz, T. et al. Aberrant PGC-1α signaling in a lamb model of persistent pulmonary hypertension of the newborn. Pediatr Res (2024). https://doi.org/10.1038/s41390-024-03223-2

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