Gene Expression Analysis in Four Dogs With Canine Pemphigus Clinical Subtypes Reveals B Cell Signatures and Immune Activation Pathways Similar to Human Disease

Abstract

Pemphigus is a group of autoimmune-mediated mucocutaneous blistering diseases characterized by acantholysis. Pemphigus has also been recognized in dogs and shares similar clinical characteristics and variants with human pemphigus. While relationships between human and canine pemphigus have been reported, gene expression patterns across species have not been described in the literature. We sought to perform gene expression analysis of lesional skin tissue from four dogs with various forms of pemphigus to examine gene expression during spontaneous disease in dogs. We found increased T and B cell signatures in canine pemphigus lesions compared to controls, as well as significant upregulation of CCL3, CCL4, CXCL10, and CXCL8 (IL8), among other genes. Similar chemokine/cytokine expression patterns and immune infiltrates have been reported in humans, suggesting that these genes play a role in spontaneous disease. Direct comparison of our dataset to previously published human pemphigus datasets revealed five conserved differentially expressed genes: CD19, WIF1, CXCL10, CD86, and S100A12. Our data expands our understanding of pemphigus and facilitates identification of biomarkers for prediction of disease prognosis and treatment response, which may be useful for future veterinary and human clinical trials.

Using Gene Expression Analysis to Understand Complex Autoimmune Skin Disease Patients: A Series of Four Canine Cutaneous Lupus Erythematosus Cases

“…We chose NanoString technology to analyze RNA from these FFPE samples because the technology works well for fragmented RNA and avoids amplification, which could introduce a data acquisition bias ( 29 ). To ascertain whether the cases presented here aligned with cases with confirmed diagnoses, we performed hierarchical clustering using ClustVis with GSE160260 [canine DLE; dataset originally published in Garelli et al ( 18 )] and GSE171079 [canine pemphigus including pemphigus erythematosus; dataset originally published in Raef et al ( 20 )]. Hierarchical clustering of gene expression for the whole panel differentiates between healthy controls and all cases, with a significant overlap between DLE and the cases presented here ( Supplementary Figures 1A,B ).…”

“…RNA was isolated from 30 µm tissue curls using the Qiagen formalin-fixed paraffin-embedded (FFPE) tissue RNeasy kit per the manufacturer's directions and as previously described (18)(19)(20). RNA was quantified using a NanoDrop spectrophotometer.…”

Microarray Gene Expression Analysis of Lesional Skin in Canine Pemphigus Foliaceus

“…In the present study, we aimed to analyze the molecular mechanisms and altered biological pathways in the skin lesions of canine PF patients. Previous studies evaluating immune markers of canine PF patients are limited to two reports [ 9 , 15 ]. In the first study, the Nanostring microarray with 160 markers was used to analyze lesional skin gene expressions from a single PF patient [ 15 ], while in the second study, qRT-PCR was used in seven canine PF skin tissues to evaluate changes in only 20 immune markers [ 9 ].…”

“…Previous studies evaluating immune markers of canine PF patients are limited to two reports [ 9 , 15 ]. In the first study, the Nanostring microarray with 160 markers was used to analyze lesional skin gene expressions from a single PF patient [ 15 ], while in the second study, qRT-PCR was used in seven canine PF skin tissues to evaluate changes in only 20 immune markers [ 9 ]. This study identified 420 DEGs possibly involved in the pathogenesis of canine PF skin lesion development.…”

Comparative Assessment of Efficacy of Prednisolone and Cyclosporine in Canine Pemphigus Complex

“…Cyclosporine, a calcineurin inhibitor, blocks T-cell infiltration, activation, and the subsequent release of inflammatory cytokines interleukin (IL)-2, IL-4, interferon (IFN)-γ, and tumour necrosis factor (TNF)-α [10,11]. Cyclosporine likely exerts the therapeutic effect in antibody-driven autoimmunity (e.g., pemphigus) by controlling B-cell activation through inhibition of reactive T helper cells interaction with naive B cells [12,13]. The main adverse reactions to cyclosporine therapy are renal dysfunction, hypertension, tremor, hirsutism, and gingival hyperplasia.…”