Authors:

Sjgrens Disease Reese Robinson

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About Paper:

Sjégren's disease (SjD) is a chronic autoimmune disorder characterized by immune-mediated destruction of salivary glands, leading to irreversible hyposalivation and significantly reduced quality of life. Current therapies do not address regeneration of damaged secretory tissue, despite evidence that salivary glands retain regenerative capacity through epithelial plasticity of ductal, acinar, and myoepithelial cells (Rocchi et al. 2021). This project identifies small molecules that promote differentiation of acinar and myoepithelial cells (MECs) to restore salivary gland function in SjD patients. Candidate compounds were identified through a high-throughput RASL-sequencing screen of 1,800 small molecules in mouse salivary gland organoids, measuring expression of acinar marker Aquaporin 5 (Aqp5) and MEC marker Calponin 1 (Cnn1). Validated hits were tested in both 2D and 3D mouse salivary gland cell culture systems across multiple doses using RT-qPCR and immunofluorescence imaging. Additionally, 17 structural derivatives of a lead compound were synthesized via flow chemistry and evaluated for biological activity. Treatment with Compounds A, B, and C significantly upregulated Cnn1 and Aqp5 expression in both 2D and 3D culture systems in a dose-responsive manner. Immunofluorescence confirmed increased numbers of Cnn1-positive cells localized to the outer edges of organoids, consistent with proper MEC positioning. Aquaporin 5 upregulation was variable across compounds; Compounds 3, 6, 12, and 16, derivatives of the lead compound, showed properly localized Aqp5 expression at the lumen membrane. Overall, 5 of 17 synthesized derivatives successfully upregulated acinar or MEC markers. These findings demonstrate that small molecules can promote salivary gland cell differentiation in vitro, supporting a regenerative strategy that complements existing immune-targeted SjD therapies.

Source:

University of Chicago

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Co-authors:

Sjgrens Disease Reese Robinson