WIP 62 - Podocyte senescence modulates progression of chronic kidney disease in neonatal hyperoxia exposure

Publication Number: WIP 62.7384

Nicholas Khuu, University of Miami Leonard M. Miller School of Medicine, Miami, FL, United States; Jo Duara, University of Miami Leonard M. Miller School of Medicine, Miami, FL, United States; Alessia Fornoni, Peggy and Harold Katz Family Drug Discovery Center, Miami, FL, United States

Background: Supplemental oxygen is an essential treatment in the care of neonates. However, it is known that continuous exposure to supplemental oxygen in preterm infants can lead to bronchopulmonary dysplasia (BPD). Previous studies have demonstrated that cellular senescence contributes to the progression of BPD (Jiang et al., 2024) and that hyperoxia treatment induces glomerular damage (Jing et al., 2015). Given that individuals born preterm have a 2-fold risk of developing chronic kidney disease (Crump et al., 2015) and hyperoxia treatment can lead to BPD and renal toxicity, we are interested in investigating the cellular mechanism of neonatal hyperoxia exposure in the propensity to developing chronic kidney disease.

Objective: We hypothesize that cellular senescence is the mechanism that underlies CKD risk in preterm with chronic supplemental hyperoxia treatment. Given that CKD is a potential long-term manifestation of early podocyte damage, our goal is to characterize the molecular changes that perpetuate cell damage and to translate our findings with clinically relevant therapeutic interventions for risk reduction.

Design/Methods: To study the effect of hyperoxia injury in the kidney, we used podocytes as an in vitro model to characterize kidney pathology. Experimentally, we defined hyperoxia treatment as 85% O2, with a positive control treatment, puromycin aminoglycoside, known to induce podocyte senescence (Pereira et al., 2023). We will subject undifferentiated human podocytes to hyperoxia treatment and PAN treatment and conduct a variety of downstream readouts to characterize senescence. First, we optimized high-throughput nuclear imaging with OPERA Phenix imaging to study changes in nuclear morphology. Secondly, we complemented these studies with immunoblot analysis of senescence markers, including p21, p16, p53, and p27. Further, we conducted Beta-galactosidase staining of treated cells to characterize senescence. Together, these preliminary studies, estimated to be completed by March 2025, will provide evidence that hyperoxia treatment induces podocyte senescence.