Faculty mentor/PI email address
pestovdg@rowan.edu
Keywords
hypoxia, ribosome, RNA, RNase L, ROS, ischemia/reperfusion injury
Date of Presentation
5-6-2026 12:00 AM
Embargo Period
5-6-2027
Poster Abstract
Ischemia/Reperfusion Injury (IRI) is a primary driver of morbidity in myocardial infarction, stroke, and acute kidney disease. Despite its clinical impact, effective therapies remain limited, necessitating the investigation of alternative pathological mechanisms and biomarkers. During IRI, metabolic dysregulation and reactive oxygen species (ROS) production damage vital macromolecules, including ribosomal RNA (rRNA). Because rRNA is highly susceptible to oxidative stress and inflammatory endonucleases like RNase L, it serves as a sensitive indicator of cellular distress. Using Northern blot analysis, we compared rRNA fragmentation in A549 cells subjected to oxygen/glucose deprivation (OGD)—an in vitro IRI model—against other oxidative stressors such as cadmium, arsenite, and erastin. Our results revealed fragmentation patterns unique to the OGD-treated group. Furthermore, these 28S and 18S rRNA patterns were significantly altered in RNase L knockout cell lines, confirming that specific cleavage events are mediated by RNase L activation.To overcome the resolution limits of Northern blotting, we developed a next-generation sequencing (NGS) pipeline to pinpoint these breaks at nucleotide-level resolution. This approach identified unique cleavage "hot spots" induced by OGD that were distinct from other stressors. By characterizing these RNase L-dependent and independent processes, this research provides critical insights into the molecular mechanisms of IRI-induced damage. Ultimately, this work establishes a framework for developing highly specific, non-invasive rRNA-based biomarkers to improve the diagnosis and monitoring of tissue injury.
Disciplines
Cardiovascular Diseases | Medicine and Health Sciences
Distinct Ribosomal RNA Fragmentation Signatures as Potential Biomarkers for Ischemia/Reperfusion Injury
Ischemia/Reperfusion Injury (IRI) is a primary driver of morbidity in myocardial infarction, stroke, and acute kidney disease. Despite its clinical impact, effective therapies remain limited, necessitating the investigation of alternative pathological mechanisms and biomarkers. During IRI, metabolic dysregulation and reactive oxygen species (ROS) production damage vital macromolecules, including ribosomal RNA (rRNA). Because rRNA is highly susceptible to oxidative stress and inflammatory endonucleases like RNase L, it serves as a sensitive indicator of cellular distress. Using Northern blot analysis, we compared rRNA fragmentation in A549 cells subjected to oxygen/glucose deprivation (OGD)—an in vitro IRI model—against other oxidative stressors such as cadmium, arsenite, and erastin. Our results revealed fragmentation patterns unique to the OGD-treated group. Furthermore, these 28S and 18S rRNA patterns were significantly altered in RNase L knockout cell lines, confirming that specific cleavage events are mediated by RNase L activation.To overcome the resolution limits of Northern blotting, we developed a next-generation sequencing (NGS) pipeline to pinpoint these breaks at nucleotide-level resolution. This approach identified unique cleavage "hot spots" induced by OGD that were distinct from other stressors. By characterizing these RNase L-dependent and independent processes, this research provides critical insights into the molecular mechanisms of IRI-induced damage. Ultimately, this work establishes a framework for developing highly specific, non-invasive rRNA-based biomarkers to improve the diagnosis and monitoring of tissue injury.