Salivary Organoids Contain More Diverse Epithelial and Less Mesenchymal Cells Than Adherent Cultures: Choose Your Sjögren’s Disease Research Tool Carefully

Ting Yang¹, Abel Soto Gamez¹, Janneke Terpstra¹, Hendrika Bootsma², Arjan Vissink¹, Frans Kroese³ and Sarah Pringle³, ¹UMCG, Groningen, Netherlands, ²Department of Rheumatology and Clinical Immunology, University of Groningen, University Medical Center Groningen, Groningen, Groningen, Netherlands, ³University Medical Center Groningen, Groningen, Netherlands

Meeting: ACR Convergence 2024

Keywords: Sjögren's syndrome, Statistical methods

Session Information

Date: Sunday, November 17, 2024

Title: Sjögren's Syndrome – Basic & Clinical Science Poster I

Session Type: Poster Session B

Session Time: 10:30AM-12:30PM

Background/Purpose: The majority of the critical pieces of work probing the involvement of the salivary gland (SG) epithelium in Sjogren’s Disease (SjD) employ salivary gland epithelial cell cultures (SGECs) presumed to recapitulate the involvement of the striated ducts in disease pathology. In recent years, organoid technology has offered a more advanced model system for studying epithelial cells in health and disease. Organoids are widely believed to accurately mimic the organization and structure of the organ from which they are derived. Organoids from the salivary gland have been described and employed in the study of SG pathology in SjD¹. We aim to investigate cells types contained in adherent SGECs and 3-dimensional SG organoid (SGO) cultures.

Methods: Matched SGEC and SGO cultures were derived from healthy parotid SGs. Bulk RNA sequencing analysis and deconvolution was performed.

Results: No striking difference in SG ductal, acinar or myoepithelial marker gene expression between SGECs and SGOs was observed. Extracellular matrix organization pathways were highly expressed in SGEC cultures, and epithelial development pathways in SGOs. Mesenchymal marker genes, including CD90, COL15A1, COL1A2, and DCN, were significantly higher expressed in SGEC cultures. Deconvolution suggested that SGECs and SGOs are heterogeneous cultures, both containing basal, striated, Krt19⁺ ductal, Ascl3⁺ ductal, intercalated, Gstt1⁺ male and female, and granulated convoluted tubule ductal cells (Figure 1). SGO cultures contain slightly more total ductal cells (46%) compared to SGECs (39%) (Figure 1). SGECs and SGOs both contain seromucous, mucous and Smgc⁺ acinar cells. SGOs also contained Bpifa2⁺ acinar cells. Mesenchymal cells likely comprise 10% of SGEC cultures, compared to 3% in SGOs (Figure 1).

Conclusion: SGECs and SGO cultures are heterogenous in nature. SGOs contain more epithelial cell types and a higher proportion of ductal cells. In contrast, mesenchymal cells comprise a larger proportion of SGECs.

Figure 1. Deconvolution of bulk RNASeq data suggests heterogenous ductal and acinar cell content of SGOs and SGECs, with more variation in epithelial cell content in SGOs, and presence of more mesenchymal cells in SGEC cultures compared to SGOs. A) Deconvolution of bulk RNA sequencing using the Cibersortx platform. Analysed samples are named on the X axes, the relative percentage of each determined cell type on the Y axes, given in the indicated colour from the key. B) Mean percentages (red text in boxes) of all detected cell types in the 4 matched SGEC and SGO cultures, represented as pie charts. C) Combined acinar and ductal cell types, mesenchymal cells and mitotic cells, as a percentage in total cells. In the case of SGECs and SGOs, values represent mean, for PG sample value is single biological sample. Key represents percentage scale. D) Statistical analysis of individual cell types in SGECs. Bars represent S.D. * = p< 0.05, ** = p<0.01.

Disclosures: T. Yang: None; A. Soto Gamez: None; J. Terpstra: None; H. Bootsma: Argenx, 2, AstraZeneca, 2, 12, Funding, Bristol Myers Squibb, 2, 12, Funding, Galapagos, 2, Novartis, 2, 12, Funding, Roche, 2, 12, Funding, Sanofi, 2, UCB, 2; A. Vissink: None; F. Kroese: None; S. Pringle: None.

To cite this abstract in AMA style:

Yang T, Soto Gamez A, Terpstra J, Bootsma H, Vissink A, Kroese F, Pringle S. Salivary Organoids Contain More Diverse Epithelial and Less Mesenchymal Cells Than Adherent Cultures: Choose Your Sjögren’s Disease Research Tool Carefully [abstract]. Arthritis Rheumatol. 2024; 76 (suppl 9). https://acrabstracts.org/abstract/salivary-organoids-contain-more-diverse-epithelial-and-less-mesenchymal-cells-than-adherent-cultures-choose-your-sjogrens-disease-research-tool-carefully/. Accessed .

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