C3a Enhances the Formation of Intestinal Organoids through C3aR1

C3a is important in the regulation of the immune response as well as in the development of organ inflammation and injury. Furthermore, C3a contributes to liver regeneration but its role in intestinal stem cell function has not been studied. We hypothesized that C3a is important for intestinal repair and regeneration. Intestinal organoid formation, a measure of stem cell capacity, was significantly limited in C3-deficient and C3a receptor (C3aR) 1-deficient mice while C3a promoted the growth of organoids from normal mice by supporting Wnt-signaling but not from C3aR1-deficient mice. Similarly, the presence of C3a in media enhanced the expression of the intestinal stem cell marker leucine-rich repeat G-protein-coupled receptor 5 (Lgr5) and of the cell proliferation marker Ki67 in organoids formed from C3-deficient but not from C3aR1-deficient mice. Using Lgr5.egfp mice we showed significant expression of C3 in Lgr5+ intestinal stem cells whereas C3aR1 was expressed on the surface of various intestinal cells. C3 and C3aR1 expression was induced in intestinal crypts in response to ischemia/reperfusion injury. Finally, C3aR1-deficient mice displayed ischemia/reperfusion injury comparable to control mice. These data suggest that C3a through interaction with C3aR1 enhances stem cell expansion and organoid formation and as such may have a role in intestinal regeneration.

[1]  M. Kolev,et al.  Intracellular complement − the complosome − in immune cell regulation , 2017, Molecular Immunology.

[2]  Mia Pras-Raves,et al.  Interplay between metabolic identities in the intestinal crypt supports stem cell function , 2017, Nature.

[3]  M. Dragunow,et al.  Complement peptide C3a stimulates neural plasticity after experimental brain ischaemia , 2017, Brain : a journal of neurology.

[4]  G. Tsokos,et al.  Intracellular Activation of Complement 3 Is Responsible for Intestinal Tissue Damage during Mesenteric Ischemia , 2017, The Journal of Immunology.

[5]  H. Clevers,et al.  Regulation and plasticity of intestinal stem cells during homeostasis and regeneration , 2016, Development.

[6]  H. Clevers,et al.  Visualization of a short-range Wnt gradient in the intestinal stem-cell niche , 2016, Nature.

[7]  R. Jenq,et al.  Interleukin-22 Promotes Intestinal Stem Cell-Mediated Epithelial Regeneration , 2015, Nature.

[8]  A. van Oudenaarden,et al.  Ascl2 acts as an R-spondin/Wnt-responsive switch to control stemness in intestinal crypts. , 2015, Cell stem cell.

[9]  Diego Diez,et al.  Neuronal exosomes facilitate synaptic pruning by up-regulating complement factors in microglia , 2015, Scientific Reports.

[10]  M. Ratajczak,et al.  Complement Component 3 is Necessary to Preserve Myocardium and Myocardial Function in Chronic Myocardial Infarction , 2014, Stem cells.

[11]  H. Clevers,et al.  Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele , 2014, The EMBO journal.

[12]  F. D. de Sauvage,et al.  Lgr5+ stem cells are indispensable for radiation-induced intestinal regeneration. , 2014, Cell stem cell.

[13]  John D Lambris,et al.  Complement anaphylatoxin C3a is a potent inducer of embryonic chick retina regeneration , 2013, Nature Communications.

[14]  Faith H. Brennan,et al.  The receptor for complement component C3a mediates protection from intestinal ischemia-reperfusion injuries by inhibiting neutrophil mobilization , 2013, Proceedings of the National Academy of Sciences.

[15]  John D Lambris,et al.  Complement-triggered pathways orchestrate regenerative responses throughout phylogenesis. , 2013, Seminars in immunology.

[16]  Wuding Zhou The new face of anaphylatoxins in immune regulation. , 2012, Immunobiology.

[17]  G. Tsokos,et al.  Immunopathogenesis of ischemia/reperfusion-associated tissue damage. , 2011, Clinical immunology.

[18]  Hans Clevers,et al.  Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts , 2011, Nature.

[19]  H. Clevers,et al.  Single Lgr5 stem cells build crypt–villus structures in vitro without a mesenchymal niche , 2009, Nature.

[20]  Ming Zhao,et al.  C3a and C5a Are Chemotactic Factors for Human Mesenchymal Stem Cells, Which Cause Prolonged ERK1/2 Phosphorylation1 , 2009, The Journal of Immunology.

[21]  H. Clevers,et al.  Identification of stem cells in small intestine and colon by marker gene Lgr5 , 2007, Nature.

[22]  M. Pekny,et al.  Complement: a novel factor in basal and ischemia‐induced neurogenesis , 2006, The EMBO journal.

[23]  John D Lambris,et al.  The Proinflammatory Mediators C3a and C5a Are Essential for Liver Regeneration , 2003, The Journal of experimental medicine.

[24]  John D Lambris,et al.  A Novel Role of Complement: Mice Deficient in the Fifth Component of Complement (C5) Exhibit Impaired Liver Regeneration1 , 2001, The Journal of Immunology.

[25]  E. Mackenzie,et al.  Expression of Receptors for Complement Anaphylatoxins C3a and C5a Following Permanent Focal Cerebral Ischemia in the Mouse , 2000, Experimental Neurology.

[26]  R. Perry Ischaemia , 1997 .

[27]  H. Scott,et al.  Intestinal mucosal lesion in low-flow states. I. A morphological, hemodynamic, and metabolic reappraisal. , 1970, Archives of surgery.