Leukocyte-Specific siRNA Delivery Revealing IRF8 as a Potential Anti-Inflammatory Target
| Autor: | Mark A. Behlke, Nuphar Veiga, Meir Goldsmith, Daniel Rosenblum, Srinivas Ramishetti, Yael Diesendruck, Eran Elinav, Itai Benhar, Dan Peer |
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| Rok vydání: | 2019 |
| Předmět: |
Surface Properties
medicine.drug_class Anti-Inflammatory Agents Pharmaceutical Science 02 engineering and technology Transfection Inflammatory bowel disease Antibodies Anti-inflammatory Polyethylene Glycols Immunomodulation Mice 03 medical and health sciences RNA interference Leukocytes medicine Animals Humans RNA Small Interfering 030304 developmental biology 0303 health sciences Tumor Necrosis Factor-alpha Chemistry Interleukins Phosphatidylethanolamines Genetic Therapy Inflammatory Bowel Diseases 021001 nanoscience & nanotechnology medicine.disease Lipids In vitro Mice Inbred C57BL Disease Models Animal Cholesterol RAW 264.7 Cells Mrna level Interferon Regulatory Factors Cancer research Nanoparticles Female IRF8 0210 nano-technology |
| Zdroj: | Journal of controlled release |
| ISSN: | 0168-3659 |
| DOI: | 10.1016/j.jconrel.2019.10.001 |
| Popis: | Interferon regulatory factor 8 (IRF8) protein plays a critical role in the differentiation, polarization, and activation of mononuclear phagocytic cells. In light of previous studies, we explored the therapeutic potential of IRF8 inhibition as immunomodulatory therapy for inflammatory bowel disease (IBD). To this end, we utilized siRNA-loaded lipid-based nanoparticles (siLNPs) and demonstrated a ∼90% reduction of IRF8 mRNA levels in vitro (PV 0.0001), alongside a notable reduction in IRF8 protein. Moreover, silencing IRF8 ex vivo in splenocytes lead to a profound downregulation of IRF8 protein, followed by an immunomodulatory effect, as represented by a decrease in the secretion of TNFα, IL6 and IL12/IL23 (IL12p40) proinflammatory cytokines (PV = 0.0045, 0.0330,0.0001, respectively). In order to silence IRF8 in vivo, selectively in inflammatory leukocytes, we used siLNPs that were coated with anti-Ly6C antibodies via our recently published ASSET targeting approach. Through this strategy, we have demonstrated a selective binding of the targeted-LNPs (T-LNPs) to Ly6C + inflammatory leukocytes. Finally, an immunomodulatory effect was demonstrated in vivo in an IBD mouse model with a profound decrease of TNFα, IL6, IL12/IL23, and IL1β pro-inflammatory cytokines (n = 5, PV 0.0001,0.0001,0.0001, 0.02, respectively) and an improvement of colon-morphology as assessed by colon-length measurements and colonoscopy (PV 0.0001). Overall, using antibody-targeted siLNPs, we showed a notable reduction of IRF8 mRNA and protein and demonstrated a targeted immunomodulation therapeutic effect ex vivo and in vivo, in the DSS colitis model. We claim that a selective silencing of IRF8 in inflammatory leukocytes (such as Ly6C+) may serve as a therapeutic approach for treating inflammatory disorders. |
| Databáze: | OpenAIRE |
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