Abstract
Macrophages contribute to peripheral nerve regeneration and produce collagen VI, an extracellular matrix protein involved in nerve function. Here, we show that collagen VI is critical for macrophage migration and polarization during peripheral nerve regeneration. Nerve injury induces a robust upregulation of collagen VI, whereas lack of collagen VI in Col6a1 −/− mice delays peripheral nerve regeneration. In vitro studies demonstrated that collagen VI promotes macrophage migration and polarization via AKT and PKA pathways. Col6a1 −/− macrophages exhibit impaired migration abilities and reduced antiinflammatory (M2) phenotype polarization, but are prone to skewing toward the proinflammatory (M1) phenotype. In vivo, macrophage recruitment and M2 polarization are impaired in Col6a1 −/− mice after nerve injury. The delayed nerve regeneration of Col6a1 −/− mice is induced by macrophage deficits and rejuvenated by transplantation of wild-type bone marrow cells. These results identify collagen VI as a novel regulator for peripheral nerve regeneration by modulating macrophage function.
Similar content being viewed by others
References
Aznavoorian S, Stracke ML, Krutzsch H et al (1990) Signal transduction for chemotaxis and haptotaxis by matrix molecules in tumor cells. J Cell Biol 10:1427–1438
Bonaldo P, Braghetta P, Zanetti M et al (1998) Collagen VI deficiency induces early onset myopathy in the mouse: an animal model for Bethlem myopathy. Hum Mol Genet 7:2135–2140
Byles V, Covarrubias AJ, Ben-Sahra I et al (2013) The TSC-mTOR pathway regulates macrophage polarization. Nat Commun 4:2834
Chen P, Bonaldo P (2013) Role of macrophage polarization in tumor angiogenesis and vessel normalization: implications for new anticancer therapies. Int Rev Cell Mol Biol 301:1–35
Chen P, Cescon M, Bonaldo P (2013) Collagen VI in cancer and its biological mechanisms. Trends Mol Med 19:410–417
Chen P, Cescon M, Bonaldo P (2014) Autophagy-mediated regulation of macrophages and its applications for cancer. Autophagy 10:192–200
Chen P, Cescon M, Megighian A, Bonaldo P (2014) Collagen VI regulates peripheral nerve myelination and function. FASEB J 28:1145–1156
Chen P, Huang Y, Bong R et al (2011) Tumor-associated macrophages promote angiogenesis and melanoma growth via adrenomedullin in a paracrine and autocrine manner. Clin Cancer Res 17:7230–7239
Christie KJ, Webber CA, Martinez JA et al (2010) PTEN inhibition to facilitate intrinsic regenerative outgrowth of adult peripheral axons. J Neurosci 30:9306–9315
Cote SC, Pasvanis S, Bounou S, Dumais N (2009) CCR7-specific migration to CCL19 and CCL21 is induced by PGE(2) stimulation in human monocytes: involvement of EP(2)/EP(4) receptors activation. Mol Immunol 46:2682–2693
Diaz-Munoz MD, Osma-Garcia IC, Iniguez MA, Fresno M (2013) Cyclooxygenase-2 deficiency in macrophages leads to defective p110gamma PI3K signaling and impairs cell adhesion and migration. J Immunol 191:395–406
Du R, Lu KV, Petritsch C et al (2008) HIF1alpha induces the recruitment of bone marrow-derived vascular modulatory cells to regulate tumor angiogenesis and invasion. Cancer Cell 13:206–220
Dubovy P, Jancalek R, Kubek T (2013) Role of inflammation and cytokines in peripheral nerve regeneration. Int Rev Neurobiol 108:173–206
Farah MH, Pan BH, Hoffman PN et al (2011) Reduced BACE1 activity enhances clearance of myelin debris and regeneration of axons in the injured peripheral nervous system. J Neurosci 31:5744–5754
Gara SK, Grumati P, Squarzoni S et al (2011) Differential and restricted expression of novel collagen VI chains in mouse. Matrix Biol 30:248–257
Germano G, Frapolli R, Belgiovine C et al (2013) Role of macrophage targeting in the antitumor activity of trabectedin. Cancer Cell 23:249–262
Ghosh-Roy A, Wu Z, Goncharov A et al (2010) Calcium and cyclic AMP promote axonal regeneration in Caenorhabditis elegans and require DLK-1 kinase. J Neurosci 30:3175–3183
Gordon S, Martinez FO (2010) Alternative activation of macrophages: mechanism and functions. Immunity 32:593–604
Gordon S, Taylor PR (2005) Monocyte and macrophage heterogeneity. Nat Rev Immunol 5:953–964
Griffin JW, George R, Ho T (1993) Macrophage systems in peripheral nerves. A review. J Neuropathol Exp Neurol 52:553–560
Horie H, Kadoya T, Hikawa N et al (2004) Oxidized galectin-1 stimulates macrophages to promote axonal regeneration in peripheral nerves after axotomy. J Neurosci 24:1873–1880
Inserra MM, Bloch DA, Terris DJ (1998) Functional indices for sciatic, peroneal, and posterior tibial nerve lesions in the mouse. Microsurgery 18:119–124
Irwin WA, Bergamin N, Sabatelli P et al (2003) Mitochondrial dysfunction and apoptosis in myopathic mice with collagen VI deficiency. Nat Genet 35:367–371
Klominek J, Robért KH, Sundqvist KG (1993) Chemotaxis and haptotaxis of human malignant mesothelioma cells: effects of fibronectin, laminin, type IV collagen, and an autocrine motility factor-like substance. Cancer Res 53:4376–4382
Liao X, Sharma N, Kapadia F et al (2011) Kruppel-like factor 4 regulates macrophage polarization. J Clin Invest 121:2736–2749
López-Vales R, Navarro X, Shimizu T et al (2008) Intracellular phospholipase A(2) group IVA and group VIA play important roles in Wallerian degeneration and axon regeneration after peripheral nerve injury. Brain 131:2620–2631
Ma L, Dong F, Zaid M et al (2012) ABCA1 protein enhances toll-like receptor 4 (TLR4)-stimulated interleukin-10 (IL-10) secretion through protein kinase A (PKA) activation. J Biol Chem 287:40502–40512
Ma Y, Halade GV, Zhang J et al (2013) Matrix metalloproteinase-28 deletion exacerbates cardiac dysfunction and rupture after myocardial infarction in mice by inhibiting M2 macrophage activation. Circ Res 112:675–688
Mantovani A, Biswas SK, Galdiero MR et al (2013) Macrophage plasticity and polarization in tissue repair and remodeling. J Pathol 229:176–185
Mokarram N, Merchant A, Mukhatyar V et al (2012) Effect of modulating macrophage phenotype on peripheral nerve repair. Biomaterials 33:8793–8801
Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8:958–969
Mueller M, Leonhard C, Wacker K et al (2003) Macrophage response to peripheral nerve injury: the quantitative contribution of resident and hematogenous macrophages. Lab Invest 83:175–185
Namikawa K, Okamoto T, Suzuki A et al (2006) Pancreatitis-associated protein-III is a novel macrophage chemoattractant implicated in nerve regeneration. J Neurosci 26:7460–7467
Niemi JP, DeFrancesco-Lisowitz A, Roldán-Hernández L et al (2013) A critical role for macrophages near axotomized neuronal cell bodies in stimulating nerve regeneration. J Neurosci 33:16236–16248
Parrinello S, Napoli I, Ribeiro S et al (2010) EphB signaling directs peripheral nerve regeneration through Sox2-dependent Schwann cell sorting. Cell 143:145–155
Perrin FE, Lacroix S, viles-Trigueros M, David S (2005) Involvement of monocyte chemoattractant protein-1, macrophage inflammatory protein-1alpha and interleukin-1beta in Wallerian degeneration. Brain 128:854–866
Ploeger DT, van Putten SM, Koerts JA et al (2012) Human macrophages primed with angiogenic factors show dynamic plasticity, irrespective of extracellular matrix components. Immunobiology 217:299–306
Santos-Sierra S, Deshmukh SD, Kalnitski J et al (2009) Mal connects TLR2 to PI3Kinase activation and phagocyte polarization. EMBO J 28:2018–2027
Schafer M, Fruttiger M, Montag D et al (1996) Disruption of the gene for the myelin-associated glycoprotein improves axonal regrowth along myelin in C57BL/Wlds mice. Neuron 16:1107–1113
Schnoor M, Cullen P, Lorkowski J et al (2008) Production of type VI collagen by human macrophages: a new dimension in macrophage functional heterogeneity. J Immunol 180:5707–5719
Shamash S, Reichert F, Rotshenker S (2002) The cytokine network of Wallerian degeneration: tumor necrosis factor-alpha, interleukin-1alpha, and interleukin-1beta. J Neurosci 22:3052–3060
Sicari BM, Johnson SA, Siu BF et al (2012) The effect of source animal age upon the in vivo remodeling characteristics of an extracellular matrix scaffold. Biomaterials 33:5524–5533
Siconolfi LB, Seeds NW (2001) Mice lacking tPA, uPA, or plasminogen genes showed delayed functional recovery after sciatic nerve crush. J Neurosci 21:4348–4355
Spencer M, Yao-Borengasser A, Unal R et al (2010) Adipose tissue macrophages in insulin-resistant subjects are associated with collagen VI and fibrosis and demonstrate alternative activation. Am J Physiol Endocrinol Metab 299:E1016–E1027
Tofaris GK, Patterson PH, Jessen KR, Mirsky R (2002) Denervated Schwann cells attract macrophages by secretion of leukemia inhibitory factor (LIF) and monocyte chemoattractant protein-1 in a process regulated by interleukin-6 and LIF. J Neurosci 22:6696–6703
Urciuolo A, Quarta M, Morbidoni V et al (2013) Collagen VI regulates satellite cell self-renewal and muscle regeneration. Nat Commun 4:1964
Vargas ME, Watanabe J, Singh SJ et al (2010) Endogenous antibodies promote rapid myelin clearance and effective axon regeneration after nerve injury. Proc Natl Acad Sci 107:11993–11998
Vitale P, Braghetta P, Volpin D et al (2001) Mechanisms of transcriptional activation of the col6a1 gene during Schwann cell differentiation. Mech Dev 102:145–156
Wang AZ, Chen JM, Fisher GW et al (1994) Improved in vitro models for assay of rheumatoid synoviocyte chemotaxis. Clin Exp Rheumatol 12:293–299
Ydens E, Cauwels A, Asselbergh B et al (2012) Acute injury in the peripheral nervous system triggers an alternative macrophage response. J Neuroinflammation 9:176
Zhang L, Johnson D, Johnson JA (2013) Deletion of Nrf2 impairs functional recovery, reduces clearance of myelin debris and decreases axonal remyelination after peripheral nerve injury. Neurobiol Dis 54:329–338
Zhu X, Lee JY, Timmins JM et al (2008) Increased cellular free cholesterol in macrophage-specific Abca1 knock-out mice enhances pro-inflammatory response of macrophages. J Biol Chem 283:22930–22941
Acknowledgments
We are grateful to W. Giuriati for technical assistance, P. Braghetta for helping with mice and R. Wagener for providing α3(VI) collagen antibodies. This work was supported by grants from the Telethon Foundation (GGP10225 and GGP11082), the Italian Ministry of Education, University and Research (RBAP11Z3YA_003), and the University of Padua Strategic Projects. P. Chen is supported by a fellowship from the Cariparo Foundation and an ImmunoTools award providing the cytokines used in this study.
Conflict of interest
The authors declare no potential conflicts of interest.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chen, P., Cescon, M., Zuccolotto, G. et al. Collagen VI regulates peripheral nerve regeneration by modulating macrophage recruitment and polarization. Acta Neuropathol 129, 97–113 (2015). https://doi.org/10.1007/s00401-014-1369-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00401-014-1369-9