Trehalose extricates impaired mitochondrial and autophagy dysregulation in patient iPSC-derived macular corneal dystrophy disease model

• Published in: Stem cell research & therapy 2024-12, Vol.15 (1), p.464-18
• Main Authors: Nayak, Divyani, Shivakumar, Shivapriya, Shetty, Rohit, Prashanthi, K N, Ghosh, Arkasubhra, Jeyabalan, Nallathambi, Chakrabarty, Koushik
• Format: Article
• Language: English
• Subjects: Autophagy; Autophagy - drug effects; Canada; Cell Differentiation; Corneal Dystrophies, Hereditary - genetics; Corneal Dystrophies, Hereditary - metabolism; Corneal Dystrophies, Hereditary - pathology; Corneal Keratocytes - drug effects; Corneal Keratocytes - metabolism; Corneal stromal cells; Differentiation; Diseases; Ethylenediaminetetraacetic acid; Genes; Germany; Humans; Induced Pluripotent Stem Cells - metabolism; Kansas; Macular corneal dystrophy; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondrial damage; Pluripotent stem cells; Proteins; Stem cells; Trehalose - pharmacology; United States
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-024-04016-4

Patient-derived induced pluripotent stem cell (iPSCs) represents a powerful tool for elucidating the underlying disease mechanisms. Macular corneal dystrophy (MCD) is an intractable and progressive bilateral corneal disease affecting the corneal stroma due to mutation/s in carbohydrate sulfotransferase 6 (CHST6) gene. The underlying molecular mechanisms leading to MCD are unclear due to a lack of human contextual model and limited access to affected corneal stromal keratocytes (CSKs) from MCD patients. This has restricted the current treatment option for MCD to restorative corneal transplantation thereby lending itself to the use of iPSCs. induced pluripotent stem cells (iPSCs) were generated from two MCD patients and a healthy participant by senai virus based reprogramming of the peripheral mononuclear blood cells (PBMCs). The iPSCs were characterized based on the expression of pluripotent markers and formation of embryoid bodies possessing tri-lineage potential. Directed differentiation of the iPSCs to corneal stromal keratocytes (CSKs) was done via intermediate induction of neural crest cells. The iCSKs were characterized by immunocytochemistry and qPCR. Proteostat staining of the iCSKs was done to validate the disease phenotype invitro. Expression of autophagy markers in the iCSKs and JC staining were visualized by immunochemistry and live-cell imaging in trehalose treated iCSKs. We show that the MCD iPSC-derived CSKs (MCDiCSKs) exhibits impaired autophagy assessed by the profiles of autophagy-associated proteins (LAMP1, LC3II/I, p62 and Beclin-1) and mitochondrial membrane potential. Significantly higher protein aggregates in MCDiCSKs was seen compared with the control, which could be rescued upon autophagy modulation. Hence, we treated MCD-iCSKs with trehalose (autophagy inducer) and showed that it protects MCD-iCSKs from mitochondrial dysfunction and maintains autophagic degradation. Our study highlights the possible pathological mechanisms involved in MCD. We found trehalose ameliorate the impaired mitochondrial and autophagy dysregulation in patient iPSC-derived macular corneal dystrophy disease model, which could be a potential alternative for MCD management.