Abstract
IntroductionIL-17 has a putative role in the pathophysiology of Sjogren’s syndrome (SS) and has been shown to be upregulated in the salivary glands of affected individuals. Sequestration of IL-17 with Adenoviral-mediated gene therapy has previously shown a benefit upon the SS-like phenotype in the Aec1/Aec2 mouse model. We sought to understand the proteomic consequences of IL-17 sequestration in the salivary gland of this mouse model as a means of illuminating the role of IL-17 in SS-like disease.MethodsUltrasound-assisted gene transfer (UAGT) was utilized to express a fusion protein composed of the extracellular portion of the IL-17 receptor fused to fragment of crystallization (Fc) in the submandibular glands of Aec1/Aec2 mice at 8 weeks of age. After confirming expression of the fusion protein and local and systemic sequestration of IL-17, proteomic profiling was performed on submandibular glands of a treated cohort of Aec1/Aec2 animals relative to the background strain and sham-treated animals.ResultsThe most notable proteomic signatures of IL-17 sequestration on SS-like disease-related proteins were Kallikrein-related peptidases, including the putative autoantigen Klk1b22. IL-17 sequestration also notably led to an isoelectric shift, but not a molecular weight shift, of Kallikrein-1, attributed to phosphorylation.ConclusionNon-viral IL-17 sequestration gene therapy in the salivary gland is feasible and downregulates expression of a putative SS autoantigen in the Aec1/Aec2 mouse.Electronic supplementary materialThe online version of this article (doi:10.1186/s13075-015-0714-2) contains supplementary material, which is available to authorized users.
Highlights
IL-17 has a putative role in the pathophysiology of Sjogren’s syndrome (SS) and has been shown to be upregulated in the salivary glands of affected individuals
Ultrasound-assisted gene transfer of the IL-17R:fragment of crystallization (Fc) cDNA results in expression levels of the fusion protein comparable to previously reported doses of adenovirus and reduces IL-17 protein to background strain levels IL-17R:Fc expression from 1 × 107 vp of Adenovirus has previously been reported to be sufficient for gene therapy efficacy in the Aec1/Aec2 mouse model [7]
We first sought to compare IL-17R:Fc mRNA expression levels after Ultrasound-assisted gene transfer (UAGT) relative to the 1 × 107 dose as well as the higher doses (1 × 109−1 × 1010vp) that have been more commonly reported in the salivary gland gene therapy literature
Summary
IL-17 has a putative role in the pathophysiology of Sjogren’s syndrome (SS) and has been shown to be upregulated in the salivary glands of affected individuals. Sequestration of IL-17 with Adenoviral-mediated gene therapy has previously shown a benefit upon the SS-like phenotype in the Aec1/Aec mouse model. Strain performed western blot analysis of salivary gland tissue 48 hours after UAGT/IL-17R:Fc, with UAGT/Luc serving as a negative control and the background strain serving as a positive control (Fig. 1b) These experiments demonstrated that IL-17 is highly expressed in the salivary glands of the Aec1/Aec mouse relative to the background strain, and that this increase is dramatically reversed by IL-17R:Fc gene transfer. These results confirmed that our IL-17R:Fc was working as intended, and allowed us to move forward into proteomic profiling of the therapeutic effect
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