Omics Analysis of Angiogenic/Angiostatic Proteins in Fetal Lung Endothelial Cells Exposed to Room Air, O2, and O2 + Retinoic Acid + Nitric Oxide

This submission has open access
Submission ID :
ESPR452
Submission Type
Submission Topic
Abstract: :

Background: Bronchopulmonary dysplasia (BPD) in extremely low birthweight (ELBW) infants consists of interrupted lung alveolar and microvascular development. The incidence of BPD remains unaltered despite advances in respiratory care, including non-invasive ventilation, and limited oxygen (O2) use. Animal studies and clinical trials of vitamin A (retinoic acid, RetA) and Nitric Oxide (NO) as single therapies suggest possible benefits for improving lung microvascular formation. We previously showed that exposure to O2 plus NO and RetA improved in vitro endothelial cell tube formation compared to O2 alone. The molecular pathways for positive microvascular effects of RetA and NO are not understood.

Objective/Hypothesis: We hypothesize that moderate O2 exposure alters the balance of a set of angiogenic and angiostatic proteins impacting lung microvascular development and that combined treatment with RetA+NO may partially rectify this O2-induced developmental imbalance.

Methods: Gestational day 19  fetal mouse lung endothelial cells (ATCC) were cultured in Room Air (RA), O2 (0.4 FiO2), ±NO (10ppm), ±RetA (10-8) for 48 hours. A targeted protein microarray of angiogenic proteins was used to analyze 53 proteins, 32 angiogenic and 11 angiostatic , in cell lysates. Responses to treatments were analyzed using R and Python packages. Significant expression differences were identified by multivariate analysis at a FDR of 0.01.

Results: Parallel coordinate plots showed that O2 alone, O2+RetA, and O2+NO had response patterns distinct from each other and from RA. Compared to RA alone, 30 proteins were significantly altered by exposure to 0.4 FiO2. Among these 30 proteins, 19 angiogenic proteins were increased and 6 decreased; 2 angiostatic proteins were increased and 3 were decreased. Eight proteins (Table) were significantly altered by exposure to combined therapy with O2+NO+RetA compared to O2 alone. Five of the 8 proteins altered by combined therapy were also impacted by O2 alone, with all showing combined therapy reversed the O2 effect except IP10 which was further decreased by combined therapy. Three angiogenic proteins modified by combined therapy were not significantly impacted by O2 alone. 

Conclusions: O2-induced alterations in microvascular development in immature lung, and correction thereof, is a complex balancing of angiogenic and angiostatic signals. Moderate (0.4 FiO2) O2 exposure profoundly impacts a subset of vasculogenic regulatory proteins in fetal lung endothelial cells. Combined therapy with NO+RetA during 0.4 FiO2 exposure partially reverses O2 effects, and also impacts some vasculogenic proteins not changed by O2 alone. This work suggests that combined NO+RetA therapy may rebalance positive and negative angiogenic pathway signals controlling microvascular formation in developing lungs exposed to O2 by modifying a subset of proteins.

Optional insertion of tables and or figures :
If the file does not load, click here to open/download the file.
Tufts Children's Hospital at Tufts Medical Center
Southern Methodist University
Tufts Children's Hospital at Tufts Medical Center

Similar Abstracts by Type

Submission ID
Submission Title
Submission Topic
Submission Type
Corresponding Author
ESPR157
Clinical Research
Original science
Aditya Chhikara
ESPR302
Epidemiology
Original science
Natasha Jouk
ESPR74
Clinical Research
Original science
Alexandra Mazo
22 hits