Acute phase cytokines and= idb

Cytokines and Inflammatory Bowel DiseaseAlexander LindUniversity of Connecticut MCB5255 2014-04-09OverviewBackground information about cytokinesCytokines and IBDArticle 1Article 2Future studies, specific aimsCytokinesCell communication and regulationInvolved in nearly all biological processes:Embryonic developmentStem cell differentiationInflammatory responsesUsed as prognostic and therapeutic agents in human disease

http://www.genecopoeia.com/product/search/pathway/h_inflamPathway.php

Cytokine storm

http://www.cytokinestorm.com/

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3180813/Pro-inflammatory/ Anti-inflammatory cytokines

OverviewBackground information about cytokinesCytokines and IBDArticle 1Article 2Future studies, specific aimsCytokine imbalance contributes to IBD through:Macrophage, Dendritic cell recruitment and activationT-cells differentiation Crohn's disease - Th1 cell mediated responseUlcerative Colitis- Th2 cell mediated response

http://www.nature.com/nrgastro/journal/v7/n2/fig_tab/nrgastro.2009.218_F1.htmlCrohn's diseaseTh1 cell induced differentiation mediated by IL-12 and IL-23TH17 cells producing IL-17 Characterized by enhanced production of IFN- and TNF-ULCERATIVE COLITISTh2 atypical immune responseLack of increased IFN- expression (abundant in CD) rather than the elevation of IL-4 the Th2-defining cytokineClassical pro-inflammatory cytokines, such as IL-1, IL-6, and TNF-Th2 cytokines IL-10 and IL-13 play a key role, increased IL-13 production by NKT cells

http://www.clinsci.org/cs/118/0707/cs1180707f03.htmhttp://www.nature.com/nrd/journal/v5/n3/fig_tab/nrd1986_ft.htmlTNF-TNF- is up-regulated in both CD and UCMostly produced by activated macrophages Transmembrane protein, activated through proteolytic cleavage Play an important role in the pathogenesis of IBD

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2731177/Anticytokine therapiesMonoclonal anti-TNF-alpha antibodies has showed good results in both inducing and maintaining remission Do not act only by neutralizing TNF-alpha: Induce monocyte and T-cell apoptosis Antibody-dependent cell-mediated cytotoxicityUnclear why certain anti-TNF therapies are effective in IBD where as other are not

Problems with current Anti TNF- therapies Liver Injury Secondary to Anti-TNF-AlphaTherapyin Inflammatory Bowel Disease: A Case Series and Review of the Literature. Parekh et. al. 2014Incidences of serious infections andtuberculosisamong patients receiving anti-tumornecrosisfactor-therapy. Yoo et. al 2014Recurrence of activetuberculosisfollowing resumption of anti-TNF-therapyin a patient with Crohn's disease. Masia et. al. 2014Tuberculosisinfection following anti-TNFtherapyin inflammatory bowel disease, despite negative screening. Debeuckelaere et. al. 2013Immune-mediated inflammatory reactions and tumors as skin side effects of inflammatory bowel disease therapy Marzano et. al. 2014

OverviewBackground information about cytokinesCytokines and IBDArticle 1Article 2Future studies, specific aimsArticle 1Orally delivered siRNA targeting macrophage Map4k4 suppresses systemic inflammation by Aoudi et.al. 2009Investigates the potential RNA interference to decrease inflammatory response

http://silence-therapeutics.com/platform-technologies/rna-interference/

Figure 1. Experimental design

Figure 2. Orally delivered Map4k4 siRNA containing vectors can be phagocytized by macrophages and silence messenger RNA expression

Conclusion: In vitro treatment of Map4k4 siRNA can silence messenger RNA expression and inhibit LPS induced TNF production.

Figure 2. (Continued)Figure 3: What effect does Map4k4 silencing have on LPS activation of previously known pathways for regulation of Tnf- production?

Conclusion: Map4k4 is a new target for suppression of Tnf- expression, activation occurs independently of previously known pathways

Figure 4. Oral administration of Map4k4 siRNA decreases mRNA expression in lung, liver and spleen.

Figure 4. ( Continued)

Conclusion: Demonstrated that macrophages in the gut internalize orally delivered siRNA, undergo RNA interference mediated gene silencing and migrate into tissues throughout the body.

Conclusion:Map4k4-siRNA administration protects mice from LPS-induced lethality through inhibition of TNF- in macrophages

Figure 5: What is the effect on TNF- expression after Map4k4 silencing?Conclusions from article 1The results in this articles describes a new strategy for oral delivery of siRNA to weaken inflammatory responses in human disease Identification of Map4k4, a previously unknown mediator of cytokine expression. Silencing of Map4k4 in macrophages in vivo protected mice from lipopolysaccharide-induced lethality by inhibiting of TNF-

This article demonstrated promising results for anti-TNF- protein therapeutics. Development of GeRP-mediated delivery of siRNA show promising results as inflammatory decreasing agents for several disease including IBD. OverviewBackground information about cytokinesCytokines and IBDArticle 1Article 2Future studies, specific aimsArticle 2Gene silencing of TNF-alpha in a murine model of acute colitis using a modified cyclodextrin delivery system by MacCarthy et.al. 2013Investigates possibility of silencing TNF- trough RNA interference in IBD

Figure 1. Stability and delivery of siRNA siRNA has short half-life time in plasma due to nuclease degradationCyclodextrin, natural occurring oligosaccharides

http://pubs.rsc.org/en/content/articlepdf/2010/MB/C001050MFigure 2.Investigation of vector-siRNA complex stability in simulated intestinal fluids

Figure 3. Quantification of TNF- and IL-6 expression in LPS stimulated cells

Figure 4. Examination of inhibitory effect of TNF- siRNA on LPS-induced cytokine secretion and expression

Conclusion: TNF- siRNA reduced LPS induced cytokine expression of TNF- and IL-6Figure 5. Clinical response after TNF- siRNA delivery in DSS treated mice

A trends towards clinincal improvement could be observed after TNF- siRNA delivery: increased body weight, reduced colon weightFigure 6. Decreased TNF- and IL-6 expression in proximal colon after TNF- -siRNA delivery to DSS-treated mice

Conclusions from Article 2 Showed in vitro studies that delivery of TNF- siRNA could interfere with LPS induced activation of pro-inflammatory cytokines Intrarectal administration of TNF-siRNA in DSS-treated mice gave indications of: clinical improvements of IBD associated featuresLower expression of TNF- and IL-6 in proximal colon

This article demonstrated promising results of a CD-based siRNA delivery system for silencing of pro-inflammatory cytokine TNF-. Potentially used in future treatment of IBD. OverviewBackground information about cytokinesCytokines and IBDArticle 1Article 2Future studies, specific aimsFuture studies There is a problem with side effects using current therapies RNA interference silencing Mp4k4 and TNF- show promising resultsCombination of silence several genes? Other ways affecting TNF- expression?

Specific aim: To use RNA interference technique to try and silence gene expression of transcription factor LITAF known to regulate TNF- expressionReferences Genecopoeia. Cytokines and Inflammatory Response http://www.genecopoeia.com/product/search/pathway/h_inflamPathway.phpKindt TJ et.al.. Kuby Immunologi 6th edition, Figure 12.Morens DM et.al. The 1918 influenza pandemic: Lessons for 2009 and the future. Critical Care Medicine. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3180813/Osterholm et. al.. Proposed Mechanism of the Cytokine Storm Evoked by Influenza virus. New England Journal of Medicine http://www.cytokinestorm.com/cytokine_storm.htmlAudet CM et. al. Interplay between pro-inflammatory cytokines and growth factors in depressive illnesses. Cellular neuroscience. http://journal.frontiersin.org/Journal/10.3389/fncel.2013.00068/fullMelmed GY et.al. Future biologic targets for IBD: potentials and pitfalls. Nature Reviews Gastroenterology and Hepatology. http://www.nature.com/nrgastro/journal/v7/n2/fig_tab/nrgastro.2009.218_F1.htmlMonteleone G et.al. T-cell-directed therapies in inamatory bowel diseases. Clinical science. http://www.clinsci.org/cs/118/0707/cs1180707f03.htmJoshua R et.al. Evolving knowledge and therapy of inflammatory bowel disase. Nature reviews. http://www.nature.com/nrd/journal/v5/n3/fig_tab/nrd1986_ft.htmlSanchez-Munoz et.al. Role of cytokines in inflammatory bowel disease. World J Gastroenterology. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2731177/RNA interference. Silence therapeutics. http://silence-therapeutics.com/platform-technologies/rna-interference/