NOD2 Activation promotes anti-inflammatory activity of human Umbilical cord blood MSCs against Inflammatory bowel disease Prof. Kyung-Sun Kang, DVM, PhD. Adult Stem Cell Research Center Seoul National University, Seoul Korea
Acknowledgement Collaborators; Byung-Chul Lee Hyung-Sick Kim Tae-Hoon Shin Gabriel Núñez, University of Michigan, USA Jong-Hwan Park, Konyang University, Korea 2
Inflammatory Bowel Disease; Ulcerative Colitis & Crohn s Disease TNBS: Trinitrobenzene sulfonic acid DSS: Dextran Sulfate Sodium
Crohn's Disease Marked by Dramatic Changes in Gut Bacteria 12 March 2014 12:00 pm thumb_article_l/public/sn-crohns.jpg The research, which involved 668 children, shows that numbers of some beneficial bacteria in the gut decrease in Crohn's patients, while the number of potentially harmful bacteria increases. 4
Pattern recognition receptors? NOD2 receptor and it s mutation site - Toll-like receptor :13 - NOD-like receptor : 23 - IBD & NOD2 NOD2 receptor in Intestinal homeostasis
NOD2; Nucleotide-binding oligomerization domain 2 1. a member of the cytosolic Nod-like receptor (NLR) family 2. consists of two N-terminal caspase-recruiting domains (CARDs), a central nucleotide binding oligomerization domain (NOD), and a C-terminal leucine rich repeat (LRR) domain for recognition of microbial molecules. 3. NOD2 senses muramyl dipeptide (MDP), a small molecule derived from the peptidoglycan of bacterial cell wall, 4. activates the serine-threonine kinase RICK (RIP-like interacting CLARP kinase) [also known as receptorinteracting protein 2 (RIP2)] via a CARD-CARD interaction (Chen et al., 2009; Inohara et al., 2003).
NOD2 and Crohn s Disease
Proposed Mechanisms of NOD2 function in intestinal homeostasis NOD2 in transplanted cells (MSCs)??
The youngest, most primitive MS Cs NOD2 is functionally expressed in human cord blood stem cells not in the bone marrow stem cells BM- MSC UCB- MSC UCB AD BM NOD2 GAPDH ICM Fertilized egg Blastocyst prenatal postnatal hucb-stem cells had/hbm-mscs
Colitis induction and Trafficking of infused hucb-mscs 3% DSS ad libitum 3% DSS TNBS MSC Injection 0 1 7 10 14 days DSS withdrawal Sacrifice No Cells CFSE UCB CFSE UCB Negative hucb-mscs hucb-mdp
NOD2-activated hucb-mscs enhanced protective effects against chemically-induced colitis
(-) PBS UCB UCB + MDP UCB-siNOD2 + MDP Kim et. al., 2013, Gastroenterology 13
NOD2-activated hucb-mscs reduce colonic inflammation in colitis Kim st. al 2013, Gastroenterology
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NOD2-activated hucb-mscs increased the colonic infiltration of Foxp3 + cells A (-) PBS UCB UCB + MDP UCB-siNOD2+MDP Foxp3 B (-) PBS MSC MSC +MDP MSC -sinod2 +MDP C Foxp3 GAPDH Kim et. al., 2013, Gastroenterlogy
MDP-induced PGE 2 is responsible for MNCs suppression by hucb-mscs COX-2 GAPDH (-) Pam 3 LPS T-DAP MDP Kim et. al., 2013, Gastroenterology
Immunosuppressive effect of hucb-mscs through NOD2-Rip2 dependent pathway COX-2 GAPDH Kim et. al., 2013, Gastroenterology
COX-2 GAPDH 19
MDP-induced PGE 2 via NOD2-RIP2 signaling increased IL-10 production and regulatory T cell population in hmncs Kim et. al., 2013, Gastroenterology
Multiple Action Mechanisms of human UCB-MSCs in Colitis microflora Colitis T cell Foxp3 - CD4 + NOD2 IL6 APC T cell Stem cells PGE2 MNC IFNg, TNFa IL-10 Th1/Th17 IL17 IL22 Colitis +UCB-MSCs
Phase 1/2a Clinical Trial for Crohn s Disease BLA PHASE 3 PHASE 2b PHASE 1&2a IND 2013. 06 IND approval (by MFDS) Early Market Advance Plan to obtain Conditional Approval as an orphan drug once phase 2b clinical trial is completed. 22
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