Performing Department
Veterinary Medicine
Non Technical Summary
Mesenchymal stem cells (MSCs) are a promising cell therapy for treating inflammatory and immune-mediated diseases, due to their secretion of proteinsthat modulate immune responses and support endogenous tissue repair. However, clinical trials investigating MSC therapy for inflammatory bowel disease (IBD) have yielded inconsistent results with some patients achieving short-term remission and others experiencing no improvement in clinical signs. Gaps in knowledge concerning the mechanisms by which the cells promote tissue regeneration and an inability to screen cells for qualitycurrently hinder MSC therapy for IBD. Priming MSCs with cytokinesto increase production of paracrine factors may enhance MSC therapeutic efficacy and help identify predictive therapeutic markers. Our hypothesis is that MSCs primed with IL-1b or TGF-b + IL-1b will have greater immunomodulatory and regenerative abilities than naïve MSCs and that prosteglandin E2 expression, which is enhanced in primed MSCs, can be used to predict therapeutic efficacy. The outcome of these studies will be an understanding of how priming MSCs affects the regenerative properties of MSCs and if expression of secreted protein can be predictive of MSC efficacy. Identifying one or more biomarkers that are predictive of successful MSC clinical therapy would reduce variability and standardize MSC regenerative therapies.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Goals / Objectives
The major goals of this project are to determine how priming mesenchymal stem cells with IL-1beta or IL-1beta + TGF-beta alters expression of paracrine factors in the stem cellsand which of these paracrine factors can be used to predict clinical efficacy for immunomodulation and regenerative capabilities.
Project Methods
We will treat human MSCs with IL-1b or TGF-b + IL-1b and perform functional assays to compare the primed MSCs to naïve MSCs. We will also determine if expression of certain paracrine factors positively correlates with the immunomodulatory and regenerative properties of the MSCs.Objective 1: Characterize paracrine factor expression in IL-1b-primed and dual TGF-b + IL-1b-primed human mesenchymal stem cells. We will analyze expression of three major paracrine factors secreted by human MSCs, PGE2, TGF-b1, and IDO, to determine how each priming strategy changes the secretion of these important regenerative cytokines. Priming will be performed three times for each donor to establish if the expression levels of these paracrine factors are consistent for each donor after priming, which is important for developing a reliable clinical therapy.Objective 2: Determine how primed MSCs modulate macrophage inflammation. We will co-culture MSCs with inflammatory (M1) macrophages and then measure changes in macrophage gene expression. PGE2, TGF-b, and IDO signaling will be inhibited in these assays to determine how these pathways promote downregulation of inflammatory cytokines from the M1 macrophages and if paracrine factor expression from the MSCs can be used to predict immunomodulation by MSCs.Objective 3: Identify how primed MSCs promote epithelial healing. MSCs will be co-cultured with "wounded" epithelial cells to compare how naïve and primed MSCs influence epithelial migration and healing. PGE2, TGF-b, and IDO signaling will again be inhibited to determine how these pathways are involved in promoting epithelial migration by MSCs and if paracrine factor expression from the MSCs can be used to predict their regenerative capabilities.Target audiences will be reached by presenting the data at scientific conferences and workshops and publishing the data in appropriate peer reviewed journals.