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Abstract Number: 0085

Fibroblasts Promote Upregulation of Cannabinoid Type 2 Receptor on Inflammatory Cells in Dermatomyositis

DeAnna Diaz1, Muhammad Bashir1, Rohan Dhiman2, Avital Baniel1, Julianne Kleitsch1, Rachita Pandya1, Meena Sharma1, Thomas Vazquez1, Ming-Lin Liu2, Mariko Momohara2 and Victoria Werth3, 1Philadelphia VAMC, Philadelphia, PA, USA and Department of Dermatology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, 2University of Pennsylvania, Philadelphia, PA, 3University of Pennsylvania and Corporal Michael J. Crescenz VAMC, Philadelphia, PA

Meeting: ACR Convergence 2023

Keywords: autoimmune diseases, cytokines, Dermatology, dermatomyositis, Fibroblasts, Dermal

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Session Information

Date: Sunday, November 12, 2023

Title: (0066–0095) T Cell Biology & Targets in Autoimmune & Inflammatory Disease Poster

Session Type: Poster Session A

Session Time: 9:00AM-11:00AM

Background/Purpose: Dermatomyositis (DM) is a chronic, systemic autoimmune disease affecting the skin, muscle, and lungs. The activation of CB2R has been shown to reduce several, key proinflammatory cytokines implicated in DM. Our group has previously shown that CB2R expression was significantly increased in in DM skin compared to blood, possibly due to increased production of local inflammatory cytokines. CB2R stimulation alleviates inflammatory response by down-regulating the expression of TNF-α, IL31, IFN-γ, and type I interferon expression in the early stage of inflammation and reducing the infiltrated inflammatory cells. Having more CB2R on inflammatory cells will lead to enhanced effects of a CB2R agonist. We hypothesized that fibroblasts (FBs) in the skin may play an important role in upregulating CB2R on inflammatory cells within the skin. Dermal fibroblasts produce and organize the extracellular matrix of the dermis. They also communicate with each other and other cell types. Therefore, we sought to examine the role of FBs in CB2R activation.

Methods: DM PBMCs, which typically have minimal CB2R expression, were cultured with medium with or without human FB supernatant. PBMCs from DM patients were cultured for 24 hrs with control medium at 1:1 dilution with or without fibroblast supernatant retrieved from a 6 well plate of 40-50% confluent primary fibroblast. We then utilized multiplexed flow cytometry to further analyze the expression of CB2R and inflammatory cytokines on 12 cell lineages.

Results: When comparing PBMCs cultured with and without FB supernatant, cells cultured with FB supernatant had a significant increase in CB2R expression on CD4+ T cells (p < 0.0001), monocyte-derived dendritic cells (moDCs) (p < 0.0001), classical Monocytes (CM) (p < 0.001), and M1 Macrophages (p < 0.0001). (Figure 1a) There was also a significant increase in inflammatory cells cytokines. When comparing PBMCs cultured with and without FB supernatant, cells cultured with FB supernatant had a significant increase in intracellular IFNγ and IFNβ expression in CD4+ T cells (p < 0.01);p < 0.001), CD8+T cells (p < 0.001; p< 0.001 ), CD11c+ cells (p < 0.001; p< 0.0001), moDCs in IFNγ (p < 0.0001), CMs (p < 0.0001; p< 0.001), and macrophages (p < 0.01). (Figure 2a-b). There was also a significant increase in inflammatory cells cytokines such as ΤΝFα, IL31 and IL4 in CD4+ T cells, moDCs, CD11c+ myeloid dendritic cells, CMs and CD68+ macrophages (data not shown).

Conclusion: These data suggest fibroblasts play a role in increasing CB2R on inflammatory cells, as well as stimulate production of cytokines in DM skin. This suggests that drugs that activate CB2R, leading to downregulation of proinflammatory cytokines, may have enhanced efficacy in the skin due to FB-induced local upregulation of CB2R on inflammatory cells in the skin.

Supporting image 1

Figure 1. (a) Flow cytometric analysis showing % of CB2R when PBMCS were cultured with fibroblast supernatant. (b) Representative figure in a single DM pt. demonstrates a higher percentage of CB2R on PBMCs cultured with fibroblast supernatant than control.

Supporting image 2

Figure 2. Flow cytometric analysis showing % of IFNγ and IFNβ when PBMCS were cultured with fibroblast supernatant.


Disclosures: D. Diaz: None; M. Bashir: None; R. Dhiman: None; A. Baniel: None; J. Kleitsch: None; R. Pandya: None; M. Sharma: None; T. Vazquez: None; M. Liu: None; M. Momohara: None; V. Werth: Abbvie, 2, Amgen, 2, 5, AnaptysBio, 2, Argenx, 5, AstraZeneca, 2, Biogen, 2, 5, Bristol Myers Squibb, 2, Celgene, 2, 5, Corbus, 5, CSL Behring, 2, 5, EMD Serono, 2, Galderma, 2, Genentech/Roche PI, 5, Gilead, 2, 5, GSK, 2, Horizon Therapeutics, 5, Janssen, 2, Kyowa Kirin, 2, Lilly, 2, Merck, 2, Nektar, 2, Novartis, 2, Octapharma, 2, Pfizer, 2, 5, Regeneron, 5, Rome Therapeutics, 2, 5, Sanofi, 2, Ventus, 5, Viela, 2, 5, Xencor, 2.

To cite this abstract in AMA style:

Diaz D, Bashir M, Dhiman R, Baniel A, Kleitsch J, Pandya R, Sharma M, Vazquez T, Liu M, Momohara M, Werth V. Fibroblasts Promote Upregulation of Cannabinoid Type 2 Receptor on Inflammatory Cells in Dermatomyositis [abstract]. Arthritis Rheumatol. 2023; 75 (suppl 9). https://acrabstracts.org/abstract/fibroblasts-promote-upregulation-of-cannabinoid-type-2-receptor-on-inflammatory-cells-in-dermatomyositis/. Accessed .
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