Unveiling senescence-associated ocular pathogenesis via lacrimal gland organoid magnetic bioassembly platform and HMGB1-Box A gene therapy

• Published in: Scientific reports 2024-09, Vol.14 (1), p.21784-13, Article 21784
• Main Authors: Ferreira, Joao Nuno, Bhummaphan, Narumol, Chaisuparat, Risa, Van Phan, Toan, Oo, Yamin, Jaru-ampornpan, Pimkwan, Matangkasombut, Oranart, Mutirangura, Apiwat
• Format: Article
• Language: English
• Subjects: 631/61; 631/80; 639/166/985; 692/699/3161; Aging; Animals; Aquaporin 5; Bioprinting; Cell death; Cellular Senescence; Deoxyribonucleic acid; DNA; DNA Damage; Dry eye disease; Dry Eye Syndromes - metabolism; Dry Eye Syndromes - pathology; Dry Eye Syndromes - therapy; Etoposide; Exocrine glands; Eye diseases; Gene therapy; Genetic Therapy - methods; HMGB1 protein; HMGB1 Protein - genetics; HMGB1 Protein - metabolism; Humanities and Social Sciences; Humans; Lacrimal Apparatus - metabolism; Lacrimal Apparatus - pathology; Lacrimal gland; Lacrimal gland and Nasolacrimal duct; Lysozyme; Metabolomics; multidisciplinary; Organoid; Organoids; Organoids - metabolism; Pathogenesis; Proteomics; Science; Science (multidisciplinary); Senescence; Swine; Transcriptomics; Transfection
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-73101-8

Dry eye disease (DED) is a multifactorial aging disorder leading to tear film insufficiency and instability. Yet, an important knowledge gap lingers in understanding senescence-associated ocular pathogenesis, due to limited in vitro translational lacrimal gland (LG) models. Consequently, this remains a major roadblock to discover effective therapies for the restoration of tear film secretion. Herein, the authors reported the magnetic bioassembly of two LG organoid platforms to recapitulate functional and aging states. Using a proof-of-concept approach, porcine primary LG cells were assembled into organoids via a magnetic 3D bioprinting (M3DB) platform. This platform could form reproducible LG organoids with epithelial hallmarks (AQP5+) and exhibit epithelial secretory functions (lysozyme activity). DNA damage-induced senescence and cell death was induced with etoposide, and LG organoid hypofunction and senescence-associated pathogenesis were observed. To confer DNA protection against aging, a novel gene therapy with Box A domain of high-mobility group box-1 (HMGB1-Box A) previously established by our group, was applied here to prevent LG cellular senescence for the first time. HMGB1-Box A transfection prevented LG organoids from senescence-associated pathogenesis at the transcriptomic, metabolomic and proteomic levels. Thus, M3DB platforms could generate functional and DNA damage-induced senescence LG organoids, and this latter damage could be prevented with HMGB1-Box A gene therapy.