advertisement
Abstract #121035 Published in IGR 25-1
RNA-seq transcriptomic profiling of TGF-β2-exposed human trabecular meshwork explants: Advancing insights beyond conventional cell culture models
Buffault J; Reboussin É; Blond F; Guillonneau X; Bastelica P; Kessal K; Akkurt Arslan M; Melik-Parsadaniantz S; Réaux-Le Goazigo A; Labbé A; Brignole-Baudouin F; Baudouin CExperimental Cell Research 2024; 442: 114220
Primary open-angle glaucoma (POAG), a leading cause of irreversible vision loss, is closely linked to increased intraocular pressure (IOP), with the trabecular meshwork (TM) playing a critical role in its regulation. The TM, located at the iridocorneal angle, acts as a sieve, filtering the aqueous humor from the eye into the collecting ducts, thus maintaining proper IOP levels. The transforming growth factor-beta 2 (TGF-β2) signaling pathway has been implicated in the pathophysiology of primary open-angle glaucoma POAG particularly, in the dysfunction of the TM. This study utilizes human TM explants to closely mimic in vivo conditions, thereby minimizing transcriptional changes that could arise from cell culture enabling an exploration of the transcriptomic impacts of TGF-β2. Through bulk RNA sequencing and immunohistological analysis, we identified distinct gene expression patterns and morphological changes induced by TGF-β2 exposure (5 ng/ml for 48 h). Bulk RNA sequencing identified significant upregulation in genes linked to extracellular matrix (ECM) regulation and fibrotic signaling. Immunohistological analysis further elucidated the morphological alterations, including cytoskeletal rearrangements and ECM deposition, providing a visual confirmation of the transcriptomic data. Notably, the enrichment analysis unveils TGF-β2's influence on both bone morphogenic protein (BMP) and Wnt signaling pathways, suggesting a complex interplay of molecular mechanisms contributing to TM dysfunction in glaucoma. This characterization of the transcriptomic modifications on an explant model of TM obtained under the effect of this profibrotic cytokine involved in glaucoma is crucial in order to develop and test new molecules that can block their signaling pathways.
Department of Ophthalmology III, Quinze-Vingts National Ophthalmology Hospital, IHU Foresight, Paris, France; Sorbonne Université, INSERM, CNRS, IHU Foresight, Institut de La Vision, Paris, France; Department of Ophthalmology, Ambroise Paré Hospital, APHP, Université de Paris Saclay, Boulogne-Billancourt, France. Electronic address: jbuffault@15-20.fr.
Full article