355 - Amphiregulin Across Development and Disease: Insights from Fetal Lung and Preterm Infants with BPD-PH

Publication Number: 1339.355

Phoebe May, Icahn School of Medicine at Mount Sinai, New York, NY, United States; Daisy Reinoso, Mount Sinai, New York, NY, United States; Hyungjoo Jeong, Icahn School of Medicine at Mount Sinai, New York, NY, United States; Christina Marshall, Icahn School of Medicine at Mount Sinai, New York, NY, United States; Megan N. Januska, Icahn School of Medicine at Mount Sinai, New York, NY, United States; Susan Kim, Seattle Children's, Seattle, WA, United States; Ya-Wen Chen, Icahn School of Medicine at Mount Sinai, New York, NY, United States; Caterina Tiozzo, Icahn School of Medicine at Mount Sinai, New York, NY, United States

Background: Amphiregulin (AREG), an EGFR ligand, orchestrates epithelial-mesenchymal signaling critical for vascular development in the lung. Disruption of this pathway in premature infants may underlie the impaired alveolarization and vascular remodeling characteristic of bronchopulmonary dysplasia (BPD)-associated pulmonary hypertension (BPD-PH), a severe pulmonary complication of prematurity. We hypothesize that diminished AREG expression links developmental vascular arrest to postnatal disease, making AREG a potential biomarker for BPD-PH.

Objective: To evaluate the developmental patterns of AREG expression and to assess AREG's potential as a biomarker for identifying preterm infants at risk for BPD-PH.

Design/Methods: To examine AREG's role in lung development and its involvement in BPD-PH, we integrated developmental and clinical approaches. Developmental nomograms of AREG expression from the embryonic to canalicular stages of lung development were determined by ELISA using both human pluripotent stem cell-derived lung organoids (hPSC-LOs) and human fetal lung tissue samples. Preterm infants at risk of BPD and BPD-PH were monitored for changes in AREG expression with serial tracheal aspirates (TAs).

Results: AREG expression increased over lung development in hPSC-LOs and their supernatant and in human fetal lung tissue (Fig. 1A-C). AREG levels in the fetal lungs were comparable to those measured in TAs from infants of similar gestational ages (Fig. 1D). Infants with BPD-PH exhibited reduced AREG levels compared to those with BPD alone, independent of BPD severity (Fig. 2A-B). This reduction was specific to BPD-PH and absent in other forms of neonatal PH, suggesting its link to acquired vasculopathy secondary to vascular remodeling (Fig. 2C). An inverse relationship between AREG and BNP - a commonly used blood biomarker for PH - was appreciated, with AREG's decline preceding BNP's rise, supporting AREG's potential utility as an early and specific biomarker of BPD-PH (Fig. 2D). Our team is currently in the process of performing immunofluorescence (IF) and FACS on fetal human lung samples and TAs to identify AREG-producing and AREG-responding (EGFR⁺) cell populations, both in lung development and the postnatal period.

Conclusion(s): AREG's role in bridging developmental and postnatal vascular remodeling suggests that it may serve as a novel biomarker and therapeutic target for predicting and managing PH in preterm infants affected by BPD.

Figure 1. AREG level increase during lung development
A: AREG expression at different time points during lung differentiation of hPSCs Lung organoids. D= days.
B: AREG expression at different time point in Lung Organoids (LO) supernatant.
C: AREG expression in Human fetal lung samples at different gestational Age. N>3 per group.
D: comparison between fetal lung samples and tracheal aspirates (TAs) of premature infants at similar gestational age (GA). Wks: weeks of gestation.

Figure2. AREG levels in TAs of BPD-PH infants are decreased compared to infants affected by uncomplicated BPD, are specific for the vascular remodeling present in BPD-PH and may reflect early PH development.
A: comparison between means AREG levels in BPD vs BPD-PH patients. BPD: patients n=8, samples n=16, BPD-PH: patients n=6 samples n=22. **** p<0.0001.
B: presence of PH affects AREG levels rather than the BPD severity. Patients n=3 each group.
* p<0.05.
C: AREG levels from TAs of BPD-PH patients compared to AREG level in TAs of infants affected by VOG (A, n=3 patients each group) and PPHN (B, n=5 patients each group). Patients were comparable for PMA.
D: comparison between AREG and BNP levels in 5 BPD-PH patients at different timepoints. PMA was expressed in weeks. * P<0.05