Autophagy is required for the development and functionality of lacrimal gland-like organoids

Lacrimal glands (LGs) serve as pivotal exocrine glands crucial for protecting the ocular surface. Dysfunction in LG cell composition or secretion is implicated in dry eye disease (DED). While autophagy plays a vital role in tissue. homeostasis in many organs, how it affects LG development and secretory function is not known. Here we have undertaken a genetic study by utilizing autophagy-deficient human embryonic stem cells and differentiating them into LG-like organoids. Autophagy-deficient LG-like organoids exhibited improper development and secretion, along with increased protein aggregation, proliferation, and cell death. These phenotypes were associated with an accumulation of PAX6, a transcription factor crucial for brain and eye development, which we identified as an autophagy substrate. Pharmacological interventions with nicotinamide mononucleotide and melatonin were able to rescue the cellular dysfunction in autophagy-deficient LG-like organoids. Together, our study highlights the role of autophagy in LG along with potential therapeutic interventions for DED. Overall design: To investigate the role of autophagy in LG-like organoids, we used multi-zonal eye cells (SEAM) differentiation from ATG5 WT and KO hESCs. We then performed gene expression profiling analysis using data obtained from bulk RNA-seq of ATG5 WT and KO hESC-derived SEAM. Comperative gene expressing profiling of RNA-seq data for ATG5 WT and KO hESC-derived SEAM.