Transplantation of IL-10-transfected endothelial progenitor cells improves retinal vascular repair via suppressing inflammation in diabetic rats
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We aimed to evaluate the effect of IL-10 gene transfection on endothelial progenitor cells (EPCs) under inflammatory conditions, and explore the therapeutic potential of IL-10-transfected EPC transplantation on nonproliferative diabetic retinopathy (NPDR).
Lentivirus vectors encoding IL-10 were constructed and introduced into EPCs isolated from rat bone marrow. After exposure to recombinant rat TNF-α, abilities of nontransfected EPCs (non-EPCs) and EPCs transfected with normal control lentivirus (EPCs-GFP) or IL-10 expressing lentivirus (EPCs-IL-10-GFP) were assessed, including migration, adhesion, and tube formation. IL-10 production by EPCs-IL-10-GFP was determined by ELISA. Following 12 weeks after establishment of diabetes, diabetic rats were randomly injected with non-EPCs, EPCs-GFP, or EPCs-IL-10-GFP via tail vein. Expression of inflammatory factors and factors associated with nuclear factor-kappa B (NF-kB) signal pathway, retinal histological analysis, and retinal vascular permeability were assessed 2 weeks after transplantation.
The detrimental effects of TNF-ɑ on the abilities of EPCs were significantly attenuated in EPCs-IL-10-GFP compared with non-EPCs and EPCs-GFP. The concentration of IL-10 in the EPCs-IL-10-GFP group was significantly higher than the non-EPCs and EPCs-GFP groups. Additionally, transplantation of EPCs-IL-10-GFP significantly inhibited inflammatory factors expression and activation of NF-kB signal pathway, improved retinal histological changes, and attenuated retinal vascular permeability.
In conclusion, transplantation of IL-10-transfected EPCs significantly improved EPCs-mediated retinal vascular repair and subsequently suppressed NPDR progression. This was associated with inflammation suppression, at least partly via inhibiting the NF-kB signal pathway. Transplantation of IL-10-transfected EPCs may be a new strategy for treatment of NPDR.
KeywordsIL-10 Endothelial progenitor cells Nonproliferative diabetic retinopathy Vascular repair Inflammation
This study was supported by Natural Science Foundation of Tianjin (Grant Numbers 12JCYBJC33900 and 14JCYBJC28000) and National Natural Science Foundation of China (Grant numbers 81371038 and 91442124).
Compliance with ethical standards
This study was supported by Natural Science Foundation of Tianjin (Grant Numbers 12JCYBJC33900 and 14JCYBJC28000) and National Natural Science Foundation of China (Grant numbers 81371038 and 91442124). The sponsor had no role in the design or conduct of this research.
Conflict of interest
All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript.
The Association for Research in Vision and Ophthalmology statement for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of Animal Care and Use Committee of Tianjin Medical University.
- 7.Long J, Wang S, Zhang Y, Liu X, Zhang H, Wang S (2013) The therapeutic effect of vascular endothelial growth factor gene- or heme oxygenase-1 gene-modified endothelial progenitor cells on neovascularization of rat hindlimb ischemia model. J Vasc Surg 58(3):756.e2–765.e2. doi: 10.1016/j.jvs.2012.11.096 CrossRefGoogle Scholar
- 10.Sen S, Merchan J, Dean J, Ii M, Gavin M, Silver M, Tkebuchava T, Yoon YS, Rasko JE, Aikawa R (2010) Autologous transplantation of endothelial progenitor cells genetically modified by adeno-associated viral vector delivering insulin-like growth factor-1 gene after myocardial infarction. Hum Gene Ther 21(10):1327–1334. doi: 10.1089/hum.2010.006 CrossRefPubMedGoogle Scholar
- 12.Brunner S, Schernthaner GH, Satler M, Elhenicky M, Hoellerl F, Schmid-Kubista KE, Zeiler F, Binder S, Schernthaner G (2009) Correlation of different circulating endothelial progenitor cells to stages of diabetic retinopathy: first in vivo data. Invest Ophthalmol Vis Sci 50(1):392–398. doi: 10.1167/iovs.08-1748 CrossRefPubMedGoogle Scholar
- 13.Brunner S, Hoellerl F, Schmid-Kubista KE, Zeiler F, Schernthaner G, Binder S, Schernthaner GH (2011) Circulating angiopoietic cells and diabetic retinopathy in type 2 diabetes mellitus, with or without macrovascular disease. Invest Ophthalmol Vis Sci 52(7):4655–4662. doi: 10.1167/iovs.10-6520 CrossRefPubMedGoogle Scholar
- 15.Feng Y, van Eck M, Van Craeyveld E, Jacobs F, Carlier V, Van Linthout S, Erdel M, Tjwa M, De Geest B (2009) Critical role of scavenger receptor-BI-expressing bone marrow-derived endothelial progenitor cells in the attenuation of allograft vasculopathy after human apo A-I transfer. Blood 113(3):755–764. doi: 10.1182/blood-2008-06-161794 CrossRefPubMedGoogle Scholar
- 18.Zimmermann O, Homann JM, Bangert A, Muller AM, Hristov G, Goeser S, Wiehe JM, Zittrich S, Rottbauer W, Torzewski J, Pfitzer G, Katus HA, Kaya Z (2012) Successful use of mRNA-nucleofection for overexpression of interleukin-10 in murine monocytes/macrophages for anti-inflammatory therapy in a murine model of autoimmune myocarditis. J Am Heart Assoc 1(6), e003293. doi: 10.1161/jaha.112.003293 CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Verma SK, Krishnamurthy P, Barefield D, Singh N, Gupta R, Lambers E, Thal M, Mackie A, Hoxha E, Ramirez V, Qin G, Sadayappan S, Ghosh AK, Kishore R (2012) Interleukin-10 treatment attenuates pressure overload-induced hypertrophic remodeling and improves heart function via signal transducers and activators of transcription 3-dependent inhibition of nuclear factor-kappaB. Circulation 126(4):418–429. doi: 10.1161/circulationaha.112.112185 CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Krishnamurthy P, Thal M, Verma S, Hoxha E, Lambers E, Ramirez V, Qin G, Losordo D, Kishore R (2011) Interleukin-10 deficiency impairs bone marrow-derived endothelial progenitor cell survival and function in ischemic myocardium. Circ Res 109(11):1280–1289. doi: 10.1161/circresaha.111.248369 CrossRefPubMedPubMedCentralGoogle Scholar
- 23.Chen YH, Lin SJ, Lin FY, Wu TC, Tsao CR, Huang PH, Liu PL, Chen YL, Chen JW (2007) High glucose impairs early and late endothelial progenitor cells by modifying nitric oxide-related but not oxidative stress-mediated mechanisms. Diabetes 56(6):1559–1568. doi: 10.2337/db06-1103 CrossRefPubMedGoogle Scholar
- 27.Carr AJ, Vugler AA, Hikita ST, Lawrence JM, Gias C, Chen LL, Buchholz DE, Ahmado A, Semo M, Smart MJ, Hasan S, da Cruz L, Johnson LV, Clegg DO, Coffey PJ (2009) Protective effects of human iPS-derived retinal pigment epithelium cell transplantation in the retinal dystrophic rat. PLoS One 4(12), e8152. doi: 10.1371/journal.pone.0008152 CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Huang H, Gandhi JK, Zhong X, Wei Y, Gong J, Duh EJ, Vinores SA (2011) TNFalpha is required for late BRB breakdown in diabetic retinopathy, and its inhibition prevents leukostasis and protects vessels and neurons from apoptosis. Invest Ophthalmol Vis Sci 52(3):1336–1344. doi: 10.1167/iovs.10-5768 CrossRefPubMedPubMedCentralGoogle Scholar