Abstract
Mesenchymal stem cells (MSCs) hold great potential for cell- and gene-based therapies for retinal degeneration. Limited survival is the main obstacle in achieving successful subretinal transplantation of MSCs. The present study sought to evaluate the effect of interleukin-13 (IL-13) gene modification on the phenotypic alteration of retinal microglia (RMG) and the survival of MSCs following subretinal grafting. In this study, LPS-activated RMG were cocultured with MSCs or IL-13-expressing MSCs (IL-13-MSCs) for 24 h, and activated phenotypes were detected in vitro. Western blotting was performed to quantify cytokine secretion by light-injured retinas following subretinal transplantation. The numbers of activated RMG and surviving grafted cells were analysed, and the integrity of the blood–retinal barrier (BRB) was examined in vivo. We found that, compared with normal MSCs, cocultured IL-13-MSCs suppressed the expression of pro-inflammatory factors and major histocompatibility complex II, promoted the expression of anti-inflammatory cytokines by activated RMG and simultaneously inhibited the proliferation of and phagocytosis by RMG. The subretinal transplantation of IL-13-MSCs increased the expression of neurotrophic factors, IL-13 and tight junction proteins in the host retina, decreased the number of phagocytic RMG and improved the survival of grafted cells. Furthermore, IL-13-MSCs alleviated BRB breakdown induced by subretinal injection. Our results demonstrate that IL-13-MSCs can polarize activated RMG to the neuroprotective M2 phenotype and enhance the survival of grafted MSCs against the damage stress induced by subretinal transplantation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- MSCs:
-
Mesenchymal stem cells
- MHC:
-
Major histocompatibility complex
- BRB:
-
Blood–retinal barrier
- RMG:
-
Retinal microglia
- IL-13:
-
Interleukin-13
- IL-13-MSCs:
-
Interleukin-13 gene-modified mesenchymal stem cells
- LV:
-
Lentivirus
- LPS:
-
Lipopolysaccharides
- RPE:
-
Retinal pigment epithelium
- TNF-α:
-
Tumour necrosis factor-α
- IL-1β:
-
Interleukin-1β
- IL-10:
-
Interleukin-10
- CNTF:
-
Ciliary neurotrophic factor
- GDNF:
-
Glial cell-derived neurotrophic factor
- ZO-1:
-
Zonula occludens-1
- SEM:
-
Scanning electron microscopy
References
Bianco P, Cao X, Frenette PS, Mao JJ, Robey PG, Simmons PJ, Wang CY (2013) The meaning, the sense and the significance: translating the science of mesenchymal stem cells into medicine. Nat Med 19:35–42. https://doi.org/10.1038/nm.3028
Damoiseaux JG, Döpp EA, Calame W, Chao D, MacPherson GG, Dijkstra CD (1994) Rat macrophage lysosomal membrane antigen recognized by monoclonal antibody ED1. Immunology 83:140–147
De Vocht N et al (2013) Quantitative and phenotypic analysis of mesenchymal stromal cell graft survival and recognition by microglia and astrocytes in mouse brain. Immunobiology 218:696–705. https://doi.org/10.1016/j.imbio.2012.08.266
Ding X, Zhang M, Gu R, Xu G, Wu H (2017) Activated microglia induce the production of reactive oxygen species and promote apoptosis of co-cultured retinal microvascular pericytes. Graefes Arch Clin Exp Ophthalmol 255:777–788. https://doi.org/10.1007/s00417-016-3578-5
Dooley D, Lemmens E, Vangansewinkel T, Le BD, Hoornaert C, Ponsaerts P, Hendrix S (2016) Cell-based delivery of interleukin-13 directs alternative activation of macrophages resulting in improved functional outcome after spinal cord injury. Stem Cell Rep 7:1099–1115. https://doi.org/10.1016/j.stemcr.2016.11.005
Hamzei TS et al (2018) Targeted intracerebral delivery of the anti-inflammatory cytokine IL13 promotes alternative activation of both microglia and macrophages after stroke. J Neuroinflamm 15:174. https://doi.org/10.1186/s12974-018-1212-7
Harhaj NS, Felinski EA, Wolpert EB, Sundstrom JM, Gardner TW, Antonetti DA (2006) VEGF activation of protein kinase C stimulates occludin phosphorylation and contributes to endothelial permeability. Invest Ophthalmol Vis Sci 47:5106–5115. https://doi.org/10.1167/iovs.06-0322
Herberg S, Shi X, Johnson MH, Hamrick MW, Isales CM, Hill WD (2013) Stromal cell-derived factor-1beta mediates cell survival through enhancing autophagy in bone marrow-derived mesenchymal stem cells. PLoS ONE 8:e58207. https://doi.org/10.1371/journal.pone.0058207
Hoornaert CJ et al (2016) In vivo interleukin-13-primed macrophages contribute to reduced alloantigen-specific T cell activation and prolong immunological survival of allogeneic mesenchymal stem cell implants. Stem Cells 34:1971–1984. https://doi.org/10.1002/stem.2360
Huang L, Xu W, Xu G (2013) Transplantation of CX3CL1-expressing mesenchymal stem cells provides neuroprotective and immunomodulatory effects in a rat model of retinal degeneration. Ocul Immunol Inflamm 21:276–285. https://doi.org/10.3109/09273948.2013.791925
Huang L, Xu G, Guo J, Xie M, Chen L, Xu W (2016) Mesenchymal stem cells modulate light-induced activation of retinal microglia through CX3CL1/CX3CR1 signaling. Ocul Immunol Inflamm 24:684–692. https://doi.org/10.3109/09273948.2015.1071405
Joe AW, Gregory-Evans K (2010) Mesenchymal stem cells and potential applications in treating ocular disease. Curr Eye Res 35:941–952
Karlstetter M, Ebert S, Langmann T (2010) Microglia in the healthy and degenerating retina: insights from novel mouse models. Immunobiology 215:685–691. https://doi.org/10.1016/j.imbio.2010.05.010
Langmann T (2007) Microglia activation in retinal degeneration. J Leukoc Biol 81:1345–1351
Le BD et al (2016) Intracerebral transplantation of interleukin 13-producing mesenchymal stem cells limits microgliosis, oligodendrocyte loss and demyelination in the cuprizone mouse model. J Neuroinflamm 13:288. https://doi.org/10.1186/s12974-016-0756-7
Li J et al (2012) Plumbagin inhibits cell growth and potentiates apoptosis in human gastric cancer cells in vitro through the NF-κB signaling pathway. Acta Pharmacol Sin 33:242–249. https://doi.org/10.1038/aps.2011.152
Lu B, Wang S, Girman S, McGill T, Ragaglia V, Lund R (2010) Human adult bone marrow-derived somatic cells rescue vision in a rodent model of retinal degeneration. Exp Eye Res 91:449–455. https://doi.org/10.1016/j.exer.2010.06.024
Mehrabadi AR, Korolainen MA, Odero G, Miller DW, Kauppinen TM (2017) Poly(ADP-ribose) polymerase-1 regulates microglia mediated decrease of endothelial tight junction integrity. Neurochem Int 108:266–271. https://doi.org/10.1016/j.neuint.2017.04.014
Mori S, Maher P, Conti B (2016) Neuroimmunology of the Interleukins 13 and 4. Brain Sci. https://doi.org/10.3390/brainsci6020018
O’Keefe GM, Nguyen VT, Benveniste EN (1999) Class II transactivator and class II MHC gene expression in microglia: modulation by the cytokines TGF-beta, IL-4, IL-13 and IL-10. Eur J Immunol 29:1275–1285. https://doi.org/10.1002/(SICI)1521-4141(199904)29:04%3c1275:AID-IMMU1275%3e3.0.CO;2-T
Orihuela R, McPherson CA, Harry GJ (2016) Microglial M1/M2 polarization and metabolic states. Br J Pharmacol 173:649–665. https://doi.org/10.1111/bph.13139
Park SS et al (2017) Advances in bone marrow stem cell therapy for retinal dysfunction. Prog Retin Eye Res 56:148–165. https://doi.org/10.1016/j.preteyeres.2016.10.002
Sanagi T et al (2010) Appearance of phagocytic microglia adjacent to motoneurons in spinal cord tissue from a presymptomatic transgenic rat model of amyotrophic lateral sclerosis. J Neurosci Res 88:2736–2746. https://doi.org/10.1002/jnr.22424
Schafer S, Calas AG, Vergouts M, Hermans E (2012) Immunomodulatory influence of bone marrow-derived mesenchymal stem cells on neuroinflammation in astrocyte cultures. J Neuroimmunol 249:40–48. https://doi.org/10.1016/j.jneuroim.2012.04.018
Sierra A, Abiega O, Shahraz A, Neumann H (2013) Janus-faced microglia: beneficial and detrimental consequences of microglial phagocytosis. Front Cell Neurosci 7:6. https://doi.org/10.3389/fncel.2013.00006
Singhal S et al (2008) Chondroitin sulfate proteoglycans and microglia prevent migration and integration of grafted Müller stem cells into degenerating retina. Stem Cells 26:1074–1082. https://doi.org/10.1634/stemcells.2007-0898
Stanzel BV et al (2014) Human RPE stem cells grown into polarized RPE monolayers on a polyester matrix are maintained after grafting into rabbit subretinal space. Stem Cell Rep 2:64–77. https://doi.org/10.1016/j.stemcr.2013.11.005
Sumi N et al (2010) Lipopolysaccharide-activated microglia induce dysfunction of the blood-brain barrier in rat microvascular endothelial cells co-cultured with microglia. Cell Mol Neurobiol 30:247–253. https://doi.org/10.1007/s10571-009-9446-7
Tzameret A et al (2014) Transplantation of human bone marrow mesenchymal stem cells as a thin subretinal layer ameliorates retinal degeneration in a rat model of retinal dystrophy. Exp Eye Res 118:135–144. https://doi.org/10.1016/j.exer.2013.10.023
Wang S et al (2010) Non-invasive stem cell therapy in a rat model for retinal degeneration and vascular pathology. PLoS ONE 5:e9200. https://doi.org/10.1371/journal.pone.0009200
Wang Y, Chen X, Cao W, Shi Y (2014) Plasticity of mesenchymal stem cells in immunomodulation: pathological and therapeutic implications. Nat Immunol 15:1009–1016. https://doi.org/10.1038/ni.3002
Xian B, Huang B (2015) The immune response of stem cells in subretinal transplantation. Stem Cell Res Ther 6:161. https://doi.org/10.1186/s13287-015-0167-1
Xu Q, Qaum T, Adamis AP (2001) Sensitive blood-retinal barrier breakdown quantitation using Evans blue. Invest Ophthalmol Vis Sci 42:789–794
Yang F, Wang D, Wu L, Li Y (2015) Protective effects of triptolide on retinal ganglion cells in a rat model of chronic glaucoma. Drug Des Dev Ther 9:6095–6107. https://doi.org/10.2147/DDDT.S92022
Zhang L, Liu H, Peng YM, Dai YY, Liu FY (2015) Vascular endothelial growth factor increases GEnC permeability by affecting the distributions of occludin, ZO-1 and tight juction assembly. Eur Rev Med Pharmacol Sci 19:2621–2627
Funding
This study was supported by grants from Startup Fund for scientific research of Fujian Medical University (Grant Number: 2016QH041), and Fund for Young and Middle-aged University Teachers’ educational research of Fujian Province (Grant Number: JT180188).
Author information
Authors and Affiliations
Contributions
HL performed and analysed majority of all experiments, including cell coculture and subretinal transplantation. XM and YJ participated in most of the experiments. HL and YY conceived and designed the experiments. The manuscript was written by HL, XM and YJ. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Ethical Approval
All animal procedures were approved by the Animal Care and Use Committee of the First Affiliated Hospital of Fujian Medical University (no.2016-YK-163) and conformed to the Association for Research in Vision and Ophthalmology (ARVO) Statement on the Use of Animals in Ophthalmic and Vision Research. All efforts were resorted to minimize the number of rats used and their suffering.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Huang, L., You, J., Yao, Y. et al. Interleukin-13 Gene Modification Enhances Grafted Mesenchymal Stem Cells Survival After Subretinal Transplantation. Cell Mol Neurobiol 40, 725–735 (2020). https://doi.org/10.1007/s10571-019-00768-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-019-00768-3