Computer aided tool for diagnosis of ENT pathologies using digital signal processing of speech and stroboscopic images.pdf
RESEARCH Open Access Complement anaphylatoxins C3a and C5a induce a failing regenerative program in cardiac resident cells. Evidence of a role for cardiac resident stem cells other than cardiomyocyte renewal David Lara-Astiaso1†, Alberto Izarra1†, Juan Camilo Estrada1, Carmen Albo1, Isabel Moscoso1, Enrique Samper1, Javier Moncayo2, Abelardo Solano2, Antonio Bernad1* and Antonio Díez-Juan1,2* Abstract Cardiac healing, which follows myocardial infarction, is a complex process guided by intricate interactions among different components. Some resident cell populations with a potential role in cardiac healing have already been described in cardiac tissues. These non-cardiomyocyte cell subsets, globally described as cardiac pluripotent/ progenitor cells (CPCs), are able to differentiate into all three major cardiac cell lineages (endothelial, smooth muscle and cardiomyocyte cells) in experimental settings. Nevertheless, physiological cardiac healing results in a fibrous scar, which remains to be fully modelled experimentally. Since a role for complement anaphylatoxins (C3a and C5a) has been described in several regeneration/repair processes, we examined the effects that C3a and C5a exert on a defined population of CPCs. We found that C3a and C5a are able to enhance CPC migration and proliferation. In vitro studies showed that this effect is linked to activation of telomerase mRNA and partial preservation of telomere length, in an NFκB-dependent manner. In addition, anaphylatoxin signalling modulates the CPC phenotype, increasing myofibroblast differentiation and reducing endothelial and cardiac gene expression. These findings may denote that C3a and C5a are able to maintain/increase the cardiac stem cell pool within the heart, whilst simultaneously facilitating and modulating resident cell differentiation. We found that this modulation was directed towards scar forming cells, which increased fibroblast/myofibroblast generation and suggests that both these anaphylatoxins could play a relevant role in the damage-coupled activation of resident cells, and regulation of the cardiac healing process after injury. Introduction The cardiac healing process is guided by intricate interactions between different components; following myocardial infarction (MI), injury, inflammation, regen- eration, and repair are all interconnected processes. It is known that these processes are inadequate and over- complicated, since certain pathological or damaging factors, such as cardiomyocyte replacement cannot be repaired. The inflammatory response represents one critical element of the cardiac healing process and is triggered by cell stress and death caused during cardiac injury. The abolition of the inflammatory response using corticosteroids not only decreases the number of infil- trating leukocytes, but also delays healing and collagen deposition (Kloner et al. 1978). Moreover it has been shown that cardiac tissue reperfusion improves overall tissue repair and that this outcome is mediated by im- proving the inflammatory reaction (Frangogiannis 2012). Over the last 30 years, the complement system has been shown to play a major role in myocardial inflammation and tissue injury following MI (Hill and Ward 1971; Walport 2001) has been experimentally inhibited at differ- ent levels, including that of anaphylatoxin (C3a, C5a) sig- nalling, to reduce ischemic injury. Experiments using * Correspondence: abernad@cnic.es; adiez@cipf.es †Equal contributors 1Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid 28029, Spain 2Vascular Repair and Regeneration Laboratory, Centro de Investigaciones, Principe Felipe, Eduardo Primo Yúfera, Valencia 46013, Spain a SpringerOpen Journal © 2012 Lara-Astiaso et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Lara-Astiaso et al. SpringerPlus 2012, 1:63 http://www.springerplus.com/content/1/1/63 animal models have shown that anaphylatoxin C5a inhib- ition protects against ischemia reperfusion (I/R) injury in diverse organs including myocardium (Vakeva et al. 1998). While anaphylatoxin C5a has been shown to be a potent activator of inflammation (Walport 2001), the upstream role of anaphylatoxin C3a has shown mixed results in ani- mal models of I/R injury. In mouse models of renal and cerebral I/R injury, C3a appears to play a key role in medi- ating inflammation (Mocco et al. 2006; Thurman et al. 2007), whereas other studies suggest it is less important (Busche and Stahl 2010; Proctor et al. 2004). Moreover, in mammals, anaphylatoxins are critical for hepatocyte proliferation and liver regeneration (Strey et al. 2003). C3a promotes homing, chemotaxis, and retention of hematopoietic stem and progenitor cells in the bone mar- row (Ratajczak et al. 2004); the anaphylatoxin receptors (C3aR, C5aR) positively regulate adult neurogenesis as well as the quantity of replacement neurons produced following cerebral ischemia (Rahpeymai et al. 2006). Re- cently, a novel and unexpected function for C3a and its receptor C3aR has also been revealed in the mutual cell-cell attraction (named co-attraction), required for maintaining cohesive clusters of migrating mesenchymal cells during early development (Carmona-Fontaine et al. 2011). The authors proposed that co-attraction and con- tact inhibition must act in concert to allow cell clusters to self-organise and respond efficiently to external signals, such as chemoattractants and repellents. There- fore anaphylatoxins seem to play both positive and nega- tive roles depending on the physiopathological context; hence their artificial modulation to improve healing still requires further research. The adult myocardium has recently been shown to harbour multipotent progenitor cells that can give rise to both myogenic and vasculogenic lineages, which can both contribute to myocardial repair (reviewed in (Barile et al. 2007; Laflamme and Murry 2011)). On the other hand, since the myocardium has a low endogenous re- generative competence, loss of a substantial amount of cardiac muscle ultimately results in scar formation. In- flammatory signals are required to guarantee the opti- mal creation of a supportive scar in the injured tissue, modulating phenotype function and gene expression in fibroblasts, endothelial cells, and leukocytes which to- gether control collagen, fibroblast/myofibroblast depos- ition and vascular network formation. An optimal inflammatory reaction leads to stable scar formation, and the low regenerative potential of cardiac tissue indicates that, at least in a physiological situation, CPCs should behave preferentially as a healing cell popu- lation that participates in scar formation, since they have a very low potential for cardiomyocyte replacement. Telomerase-competent CPCs with long telomeres are present in the atria and apex storage regions of the heart; following activation by growth factors they migrate to damaged areas, where they although have the potential to create a population of young myocytes (Itzhaki-Alfia et al. 2009) their contribution to regenerated cardiac tis- sue is very low. Experimental data suggest that the regen- erative potential of endogenous adult stem cells is low, and multiple reports show that defects in telomere maintenance impairs organ regeneration like liver (Hartmann et al. 2011) or in haematopoietic stem cell maintenance (Calado and Young 2008). In addition, telomere shortening during progenitor cell proliferation affects the function of the brain, pancreas, bone marrow, and heart, pointing to stem cell dysfunction as a critical determinant of organ aging/regeneration (Beausejour and Campisi 2006; Harrington and Greider 1991). There- fore efficient pro-regenerative signalling should modulate telomerase activation, allowing efficient cell proliferation, and also modulation of cell differentiation towards a healing phenotype. In this work we describe the role of complement anaphylatoxins in CPC biology and prove that C3a and C5a are able to enhance cell migration and proliferation. This effect is linked to telomerase mRNA activity and ac- tivation, and a partial preservation of telomere length. Conversely anaphylatoxin stimulation influences CPC fate, pushing them towards myofibroblast differentiation, reduces endothelial gene expression, and increases colla- gen and smooth muscle gene expression, thus supporting a role for them in cardiac scar formation. Hence, enhanced proliferation ability and telomere length maintenance could denote that both C3a and C5a anaphylatoxins can help to maintain the cardiac resi- dent cell pool within the heart during injury, and fa- cilitate their function as scar forming cells. Results CPC culture and characterisation Small biopsies of murine adult hearts were placed on gelatin/fibronectin plates (Figure 1A (1)). Following an ini- tial outgrowth of fibroblast-like cells, within 5–7 days of explant plating, small, round and poorly adherent cells appeared and expanded (Figure 1A (2)). These cells, called explant-derived cells (EDCs) could be detached by gently pipetting, and were harvested and cultured to form cardiospheres (Figure 1A (3)). Using immunofluorescence, EDCs were found to express the cardiac markers, Nkx2.5, Gata4, Cx43, and Mef2c (Figure 1B) and also cell surface makers c-kit and Sca-1 (Figure 1B). During cardiosphere culture, proliferative cells (Ki67 positive) were found pri- marily in the external part of the sphere, in contrast to the CPC marker c-kit predominately found in the core. Sca-1 was more homogenously expressed throughout the whole culture. EDCs showed a very low expression for the vascu- lar markers CD31 and aSMA. Lara-Astiaso et al. SpringerPlus 2012, 1:63 Page 2 of 15 http://www.springerplus.com/content/1/1/63 et al. 2002). Although clinical and biological data shows that the heart has a very low endogenous capacity for re- generation when severe damage is inflicted, multiple en- dogenous putative cardiac stem cell or progenitor cell populations have been identified and isolated. Markers, traditionally associated with blood, bone marrow or pluri- potent stem cells, have been used by several independent groups to identify these cells in adult or postnatal hearts in humans and other mammalian species (Smith et al. 2008). We have worked with a population (CPCs), as described by Messina et al. (Messina et al. 2004), this population expresses the cardiac markers, Nkx2.5, Gata4, Cx43 and Mef2c (Figure 1B) and therefore had a putative cardiogenic potential. Thus our first task was to under- stand why, if these cells have the potential to differentiate in cardiomyocytes, physiological cardiac healing usually results as a fibrotic scar and shows very low levels of cardiomyocyte replacement. We investigated the potential of these cells to differentiate to all three lineages described, and facilitated differentiation by using 10% FBS to improve myofibroblast differentiation, and EGM on Matrigel to increase endothelial lineage differentiation. Interestingly anaphylatoxins repress Gata4 expression and increase the expression of some early cardiac markers (Nkx2.5, Tbx3, Tbx5, Actc1) in CPCs. Endothelial cell (EC) lineage differ- entiation was clearly improved by culture on Matrigel using EGM media and in these conditions, stimulation with C3a and C5a anaphylatoxins reduced endothelial gene expression. In agreement with this data, immunofluores- cence also showed the absence of ECs in the presence of anaphylatoxins. Inhibition of EC differentiation is paralleled by induction of Twist and Snail genes together with Fsp1 expression. Fsp1 is one of the markers of fibro- blast formation and is a cytoskeletal protein belonging to the calmodulin-S100-troponin C superfamily of intracellu- lar calcium binding proteins associated with cytoskeletal fibres, cell motility, and mesenchymal phenotype (Zimmer et al. 1995). Therefore the expression of Snail1 and Twist1 genes, together with the significant increase in Fsp1 ex- pression, indicated that an endMT-like differentiation event might be occurring in CPCs. In addition, anaphy- latoxins induce the expression of an array of genes asso- ciated with myofibroblasts that probably abolishes their role in regeneration but hints at their relevance in the car- diac healing process that follows MI. In this scenario, com- plement anaphylatoxins would activate migration of CPCs to the damaged area, induce their proliferation and finally, upon persistence of local signalling, would promote their differentiation into myofibroblast cells that would be able to fill and quickly repair the damaged heart area, preven- ting further cardiac complications. Gata4 has been proved to be a critical early factor in cardiogenesis, it lays upstream of Tbx5, Nkx2.5, and Act1 in the cardiogenic transcription network, and acts together with Baf60c to generate a chromatin state competent for cardiogenic differentiation (Takeuchi and Bruneau 2009). Therefore C3a and C5a may block cardiomyocyte differen- tiation by repressing Gata4 expression. The cardiogenic factors Nkx2.5, Actc1 and specially Tbx3 and Tbx5 may have roles other than their cardiogenic ones, including modulating the response of CPCs to complement anaphylatoxins, for instance they might be involved in the C3a/C5a dependent myofibroblast differentiation process. It is known that several mesodermal cells (cardiomyocytes and myofibroblasts) share parts of their transcriptional dif- ferentiation networks, supporting the existence of a com- mon myocardial and smooth muscle cell precursor in the developing embryo. This is consistent with in vivo studies, which show the co-expression of numerous smooth muscle genes in myocardial progenitor cells (Wu et al. 2006). Therefore Tbx3 and Tbx5 would be common to both cell types, and Gata4 levels would be critical in modu- lating the fate decision of progenitor cells. Interestingly it has been proposed that Gata4 expression can play a role in the developmental regulation of cardiac fibroblasts and has a function in the maintenance of cardiac-resident progenitors (Jankowski 2009). This raises the hypothesis that C3a and C5a are factors that affect CPC fate by switching the balance of different lineage specific factors: they shut down the expression of endothelial and cardiac factors and induce the expression of myofibroblast factors (TCF21, SM22α and Myocardin), irreversibly pushing CPCs towards the myofibroblast fate. Taken together, these findings shed some light onto what have been described as endogenous cardiac stem cells and some mechanistic insight about the limited po- tential of CPCs to generate new cardiomyocytes after cardiac injury. We can hypothesize that anaphylatoxin release at early time after cardiac injury is able to in- crease CPC numbers and to promote migration towards the site of injury. This signal also increases CPC poten- tial to differentiate into myofibroblast lineages that would participate in scar formation. This situation gives an advantage to cardiac tissue promoting a fast healing in MI. Differentiation toward myofibroblast in CPCs would make available a higher number of myofibroblasts to contribute to other sources of myofribroblasts (Krenning et al. 2010) during the resolution stage of MI. This situation is different from physiological cardiac cell turnover. During life possibly other signals in absence of inflammatory signals will permit cardiomyocyte differen- tiation to replace exhausted cells. Interestingly recent evidence reported by Jianqin Ye et al. (2012) have shown that there is a significant increase in the proliferative capacity of CS-forming cells isolated from the “middle aged” heart following acute MI resulting in a significant rise in the number of CSs in vitro and this increase is most pronounced within the first week post-MI. In Lara-Astiaso et al. SpringerPlus 2012, 1:63 Page 13 of 15 http://www.springerplus.com/content/1/1/63 addition they show that show for the first time that the CS cells obtained from 1-week post-MI hearts engraft in ischemic myocardium and restore cardiac function at 25 days post-injection in vivo. However, we did not find evidence for differentiation of these cells into mature cardiomyocytes or new vessels althouhgt promoted angiogenesis in vivo. Their data suggest that early signals that happens after MI would commit these cells to a more healing phenotype instead of regenerative fate. To heal the heart adding new cardiomyocyte probably is a more delicate and time-consuming situation that mammalian wounded heart cannot go through in a wild environment. Thus the signals involved in cardiac healing have been selected to permit a fast healing situ- ation increasing myofibroblast differentiation but in other hand this impairs cardiomyocyte renewal. Deeper understanding of these mechanisms would help to im- prove wounded heart regeneration. Competing interests The authors indicate no potential conflicts of interests. Authors’ contributions ADJ, DLA and AI drafted the manuscript and performed Q-PCR, cell culture validation and IF experiments. CA, JCA and ES developed TRAP and FISH experiments. IM, JM and AS perform CHIP and cell culture. ADJ and AB conceived the project, read, and interpreted the results. All authors read and approved the final manuscript. Acknowledgements We thank Dr Alfonso Luque for immortalized mouse endothelial cells. All members of Diez-Juan laboratory for their helpful discussions and technical support, specialy to Virginia Zorita, Nuria Martí and Vanessa Blanca, Microscopy and Cellomics Units, at CIPF and Maria Ledran at EFL for Scientific editing. Funding This work was supported by grant to ADJ from the Ministry of Science and Innovation (SAF 2009–1000) and a MICCIN "Ramon y Cajal" contract and by a grant to ABM from GENOMA ESPAÑA (Spain) part of the MACIA research project. Received: 18 October 2012 Accepted: 30 November 2012 Published: 12 December 2012 References Amsterdam EA, Stahl GL, Pan HL, Rendig SV, Fletcher MP, Longhurst JC (1995) Limitation of reperfusion injury by a monoclonal antibody to C5a during myocardial infarction in pigs. Am J Physiol 268:H448–H457 Arslan F, de Kleijn DP, Pasterkamp G (2011) Innate immune signaling in cardiac ischemia. Nat Rev Cardiol 8:292–300 Banerjee PP, Jagadeesh S (2009) Non-radioactive assay methods for the assessment of telomerase activity and telomere length. Methods Mol Biol 523:383–394 Barile L, Messina E, Giacomello A, Marban E (2007) Endogenous cardiac stem cells. Prog Cardiovasc Dis 50:31–48 Beausejour CM, Campisi J (2006) Ageing: balancing regeneration and cancer. Nature 443:404–405 Bergmann O, Bhardwaj RD, Bernard S, Zdunek S, Barnabe-Heider F, Walsh S, Zupicich J, Alkass K, Buchholz BA, Druid H et al (2009) Evidence for cardiomyocyte renewal in humans. Science 324:98–102 Brown K, Gerstberger S, Carlson L, Franzoso G, Siebenlist U (1995) Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation. Science 267:1485–1488 Busche MN, Stahl GL (2010) Role of the complement components C5 and C3a in a mouse model of myocardial ischemia and reperfusion injury. Ger Med Sci 8, doi:pii: Doc20. PubMed PMID: 20930931; PubMed Central PMCID: PMC2940219. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2940219/ Calado RT, Young NS (2008) Telomere maintenance and human bone marrow failure. Blood 111:4446–4455 Carmona-Fontaine C, Theveneau E, Tzekou A, Tada M, Woods M, Page KM, Parsons M, Lambris JD, Mayor R (2011) Complement fragment C3a controls mutual cell attraction during collective cell migration. Dev Cell 21:1026–1037 Choi SH, Jung SY, Kwon SM, Baek SH (2012) Perspectives on stem cell therapy for cardiac regeneration. Advances and challenges. Circ J 76:1307–1312 Choudhary B, Karande AA, Raghavan SC (2012) Telomere and telomerase in stem cells: relevance in ageing and disease. Front Biosci (Schol Ed) 4:16–30 Diez-Juan A, Andres V (2003) Coordinate control of proliferation and migration by the p27Kip1/cyclin-dependent kinase/retinoblastoma pathway in vascular smooth muscle cells and fibroblasts. Circ Res 92:402–410 Frangogiannis NG (2012) Regulation of the inflammatory response in cardiac repair. Circ Res 110:159–173 Harrington LA, Greider CW (1991) Telomerase primer specificity and chromosome healing. Nature 353:451–454 Hartmann D, Srivastava U, Thaler M, Kleinhans KN, N'Kontchou G, Scheffold A, Bauer K, Kratzer RF, Kloos N, Katz SF et al (2011) Telomerase gene mutations are associated with cirrhosis formation. Hepatology 53:1608–1617 Hill JH, Ward PA (1971) The phlogistic role of C3 leukotactic fragments in myocardial infarcts of rats. J Exp Med 133:885–900 Hortelano S, Lopez-Fontal R, Traves PG, Villa N, Grashoff C, Bosca L, Luque A (2010) ILK mediates LPS-induced vascular adhesion receptor expression and subsequent leucocyte trans-endothelial migration. Cardiovasc Res 86:283–292 Huber MA, Kraut N, Beug H (2005) Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 17:548–558 Itzhaki-Alfia A, Leor J, Raanani E, Sternik L, Spiegelstein D, Netser S, Holbova R, Pevsner-Fischer M, Lavee J, Barbash IM (2009) Patient characteristics and cell source determine the number of isolated human cardiac progenitor cells. Circulation 120:2559–2566 Jankowski M (2009) GATA4, a new regulator of cardiac fibroblasts, is sensitive to natriuretic peptides. Cardiovasc Res 84:176–177 Kloner RA, Fishbein MC, Lew H, Maroko PR, Braunwald E (1978) Mummification of the infarcted myocardium by high dose corticosteroids. Circulation 57:56–63 Kohl J (2001) Anaphylatoxins and infectious and non-infectious inflammatory diseases. Mol Immunol 38:175–187 Krenning G, Zeisberg EM, Kalluri R (2010) The origin of fibroblasts and mechanism of cardiac fibrosis. J Cell Physiol 225:631–637 Laflamme MA, Murry CE (2011) Heart regeneration. Nature 473:326–335 Martinez P, Blasco MA (2011) Telomeric and extra-telomeric roles for telomerase and the telomere-binding proteins. Nat Rev Cancer 11:161–176 Messeguer X, Escudero R, Farre D, Nunez O, Martinez J, Alba MM (2002) PROMO: detection of known transcription regulatory elements using species-tailored searches. Bioinformatics 18:333–334 Messina E, De Angelis L, Frati G, Morrone S, Chimenti S, Fiordaliso F, Salio M, Battaglia M, Latronico MV, Coletta M et al (2004) Isolation and expansion of adult cardiac stem cells from human and murine heart. Circ Res 95:911–921 Mocco J, Mack WJ, Ducruet AF, Sosunov SA, Sughrue ME, Hassid BG, Nair MN, Laufer I, Komotar RJ, Claire M et al (2006) Complement component C3 mediates inflammatory injury following focal cerebral ischemia. Circ Res 99:209–217 Monsinjon T, Gasque P, Chan P, Ischenko A, Brady JJ, Fontaine MC (2003) Regulation by complement C3a and C5a anaphylatoxins of cytokine production in human umbilical vein endothelial cells. FASEB J 17:1003–1014 Pan ZK (1998) Anaphylatoxins C5a and C3a induce nuclear factor kappaB activation in human peripheral blood monocytes. Biochim Biophys Acta 1443:90–98 Proctor LM, Arumugam TV, Shiels I, Reid RC, Fairlie DP, Taylor SM (2004) Comparative anti-inflammatory activities of antagonists to C3a and C5a receptors in a rat model of intestinal ischaemia/reperfusion injury. Br J Pharmacol 142:756–764 Rahpeymai Y, Hietala MA, Wilhelmsson U, Fotheringham A, Davies I, Nilsson AK, Zwirner J, Wetsel RA, Gerard C, Pekny M, Pekna M (2006) Complement: a novel factor in basal and ischemia-induced neurogenesis. EMBO J 25:1364–1374 Ratajczak J, Reca R, Kucia M, Majka M, Allendorf DJ, Baran JT, Janowska-Wieczorek A, Wetsel RA, Ross GD, Ratajczak MZ (2004) Mobilization studies in mice deficient in either C3 or C3a receptor (C3aR) reveal a novel role for complement in retention of hematopoietic stem/progenitor cells in bone marrow. Blood 103:2071–2078 Lara-Astiaso et al. SpringerPlus 2012, 1:63 Page 14 of 15 http://www.springerplus.com/content/1/1/63 Samper E, Flores JM, Blasco MA (2001) Restoration of telomerase activity rescues chromosomal instability and premature aging in Terc−/−mice with short telomeres. EMBO Rep 2:800–807 Schraufstatter IU, Discipio RG, Zhao M, Khaldoyanidi SK (2009) C3a and C5a are chemotactic factors for human mesenchymal stem cells, which cause prolonged ERK1/2 phosphorylation. J Immunol 182:3827–3836 Shinjyo N, Stahlberg A, Dragunow M, Pekny M, Pekna M (2009) Complement- derived anaphylatoxin C3a regulates in vitro differentiation and migration of neural progenitor cells. Stem Cells 27:2824–2832 Simonsen JL, Rosada C, Serakinci N, Justesen J, Stenderup K, Rattan SI, Jensen TG, Kassem M (2002) Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells. Nat Biotechnol 20:592–596 Smith LL, Coller HA, Roberts JM (2003) Telomerase modulates expression of growth-controlling genes and enhances cell proliferation. Nat Cell Biol 5:474–479 Smith RR, Barile L, Messina E, Marban E (2008) Stem cells in the heart: what's the buzz all about?–Part 1: preclinical considerations. Hear Rhythm 5:749–757 Stampfer MR, Garbe J, Levine G, Lichtsteiner S, Vasserot AP, Yaswen P (2001) Expression of the telomerase catalytic subunit, hTERT, induces resistance to transforming growth factor beta growth inhibition in p16INK4A(−) human mammary epithelial cells. Proc Natl Acad Sci U S A 98:4498–4503 Strey CW, Markiewski M, Mastellos D, Tudoran R, Spruce LA, Greenbaum LE, Lambris JD (2003) The proinflammatory mediators C3a and C5a are essential for liver regeneration. J Exp Med 198:913–923 Sun Y, Kiani MF, Postlethwaite AE, Weber KT (2002) Infarct scar as living tissue. Basic Res Cardiol 97:343–347 Takeuchi JK, Bruneau BG (2009) Directed transdifferentiation of mouse mesoderm to heart tissue by defined factors. Nature 459:708–711 Testa L, Van Gaal WJ, Bhindi R, Biondi-Zoccai GG, Abbate A, Agostoni P, Porto I, Andreotti F, Crea F, Banning AP (2008) Pexelizumab in ischemic heart disease: a systematic review and meta-analysis on 15,196 patients. J Thorac Cardiovasc Surg 136:884–893 Thurman JM, Lenderink AM, Royer PA, Coleman KE, Zhou J, Lambris JD, Nemenoff RA, Quigg RJ, Holers VM (2007) C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ishemia/reperfusion. J Immunol 178:1819–1828 Vakeva AP, Agah A, Rollins SA, Matis LA, Li L, Stahl GL (1998) Myocardial infarction and apoptosis after myocardial ischemia and reperfusion: role of the terminal complement components and inhibition by anti-C5 therapy. Circulation 97:2259–2267 Van Beek J, Bernaudin M, Petit E, Gasque P, Nouvelot A, MacKenzie ET, Fontaine M (2000) Expression of receptors for complement anaphylatoxins C3a and C5a following permanent focal cerebral ischemia in the mouse. Exp Neurol 161:373–382 van der Pals J, Koul S, Andersson P, Gotberg M, Ubachs JF, Kanski M, Arheden H, Olivecrona GK, Larsson B, Erlinge D (2010) Treatment with the C5a receptor antagonist ADC-1004 reduces myocardial infarction in a porcine ischemia- reperfusion model. BMC Cardiovasc Disord 10:45 Walport MJ (2001) Complement second of two parts. N Engl J Med 344:1140–1144 Wu SM, Fujiwara Y, Cibulsky SM, Clapham DE, Lien CL, Schultheiss TM, Orkin SH (2006) Developmental origin of a bipotential myocardial and smooth muscle cell precursor in the mammalian heart. Cell 127:1137–1150 Ye J, Boyle A, Shih H, Sievers RE, Zhang Y, Prasad M, Su H, Zhou Y, Grossman W, Bernstein HS, Yeghiazarians Y (2012) Sca-1+ cardiosphere-derived cells are enriched for Isl1-expressing cardiac precursors and improve cardiac function after myocardial injury. PLoS One 7:e30329 Zhang H, Qin G, Liang G, Li J, Barrington RA, Liu DX (2007) C5aR-mediated myocardial ischemia/reperfusion injury. Biochem Biophys Res Commun 357:446–452 Zimmer DB, Cornwall EH, Landar A, Song W (1995) The S100 protein family: history, function, and expression. Brain Res Bull 37:417–429 doi:10.1186/2193-1801-1-63 Cite this article as: Lara-Astiaso et al.: Complement anaphylatoxins C3a and C5a induce a failing regenerative program in cardiac resident cells. Evidence of a role for cardiac resident stem cells other than cardiomyocyte renewal. SpringerPlus 2012 1:63. Submit your manuscript to a  journal and beneﬁ t from: 7 Convenient online submission 7 Rigorous peer review 7 Immediate publication on acceptance 7 Open access: articles freely available online 7 High visibility within the ﬁ eld 7 Retaining the copyright to your article     Submit your next manuscript at 7 springeropen.com Lara-Astiaso et al. SpringerPlus 2012, 1:63 Page 15 of 15 http://www.springerplus.com/content/1/1/63
Title: Complement anaphylatoxins C3a and C5a induce a failing regenerative program in cardiac resident cells. ​ Evidence of a role for cardiac resident stem cells other than cardiomyocyte renewal
Authors: David Lara-Astiaso, Alberto Izarra, Juan Camilo Estrada, Carmen Albo, Isabel Moscoso, Enrique Samper, Javier Moncayo, Abelardo Solano, Antonio Bernad, Antonio Díez-Juan
Publisher: SpringerPlus
Date: December 12, 2012 ​
Abstract: Cardiac healing, which follows myocardial infarction, is a complex process guided by intricate interactions among different components. ​ Some resident cell populations with a potential role in cardiac healing have already been described in cardiac tissues. ​ These non-cardiomyocyte cell subsets, globally described as cardiac pluripotent/progenitor cells (CPCs), are able to differentiate into all three major cardiac cell lineages (endothelial, smooth muscle, and cardiomyocyte cells) in experimental settings. ​ Nevertheless, physiological cardiac healing results in a fibrous scar, which remains to be fully modeled experimentally. ​ Since a role for complement anaphylatoxins (C3a and C5a) has been described in several regeneration/repair processes, we examined the effects that C3a and C5a exert on a defined population of CPCs. ​ We found that C3a and C5a are able to enhance CPC migration and proliferation. ​ In vitro studies showed that this effect is linked to activation of telomerase mRNA and partial preservation of telomere length, in an NFκB-dependent manner. ​ In addition, anaphylatoxin signaling modulates the CPC phenotype, increasing myofibroblast differentiation and reducing endothelial and cardiac gene expression. ​ These findings may denote that C3a and C5a are able to maintain/increase the cardiac stem cell pool within the heart, whilst simultaneously facilitating and modulating resident cell differentiation. ​ We found that this modulation was directed towards scar-forming cells, which increased fibroblast/myofibroblast generation and suggests that both these anaphylatoxins could play a relevant role in the damage-coupled activation of resident cells, and regulation of the cardiac healing process after injury. ​