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CX3CR1 deficiency accelerates the development of retinopathy in a rodent model of type 1 diabetes

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An Erratum to this article was published on 18 April 2017

Abstract

In this study, the role of CX3CR1 in the progression of diabetic retinopathy (DR) was investigated. The retinas of wild-type (WT), CX3CR1 null (CX3CR1gfp/gfp, KO), and heterozygous (CX3CR1+/gfp, Het) mice were compared in the presence and absence of streptozotocin (STZ)-induced diabetes. CX3CR1 deficiency in STZ-KO increased vascular pathology at 4 months of diabetes, as a significant increase in acellular capillaries was observed only in the STZ-KO group. CX3CR1 deficiency and diabetes had similar effects on retinal neurodegeneration measured by an increase in DNA fragmentation. Retinal vascular pathology in STZ-KO mice was associated with increased numbers of monocyte-derived macrophages in the retina. Furthermore, compared to STZ-WT, STZ-KO mice exhibited increased numbers of inflammatory monocytes in the bone marrow and impaired homing of monocytes to the spleen. The induction of retinal IL-10 expression by diabetes was significantly less in KO mice, and when bone marrow-derived macrophages from KO mice were maintained in high glucose, they expressed significantly less IL-10 and more TNF-α in response to LPS stimulation. These findings support that CX3CR1 deficiency accelerates the development of vascular pathology in DR through increased recruitment of proinflammatory myeloid cells that demonstrate reduced expression of anti-inflammatory IL-10.

Key messages

• CX3CR1 deletion in STZ-diabetic mice accelerated the onset of diabetic retinopathy (DR).

• The early onset of DR was associated with increased retinal cell apoptosis.

• The early onset of DR was associated with increased recruitment of bone marrow-derived macrophages to the retina.

• Bone marrow-derived macrophages from CX3CR1 KO diabetic mice expressed more TNF-α and less IL-10.

• The role of IL-10 in protection from progression of DR is highlighted.

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Acknowledgments

The authors would like to thank Neal Benson from the cytometry core at the ICBR in University of Florida. Research was supported by NIH grants: EY012601-15, EY007739-25, EY018358, EY023629 (MB), DK 090730 and HL110170 (MBG).

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    Authors and Affiliations

    1. Eugene and Marilyn Glick Eye Institute, Department of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA

      Eleni Beli, James M. Dominguez II, Ping Hu, Sergio Li Calzi, Tatiana E. Salazar, Yaqian Duan, Michael E. Boulton & Maria B. Grant

    2. Department of Pharmacology and Therapeutics, University of Florida, Gainesville, FL, USA

      James M. Dominguez II, Jeffrey S. Thinschmidt, Sergio Caballero, Defang Luo, Tatiana E. Salazar, Jeffrey K. Harrison & Maria B. Grant

    3. Department of Physiology, Michigan State University, East Lancing, MI, USA

      Susanna Mohr

    4. Department of Ophthalmology and Visual Sciences, University of Michigan, Kellogg Eye Center, Ann Arbor, MI, USA

      Sumathi Shanmugam & Steven F. Abcouwer

    5. Department of Ophthalmology, Duke University School of Medicine, Durham, NC, USA

      Daniel R. Saban

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    1. Eleni Beli
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    Correspondence to Maria B. Grant.

    Additional information

    Eleni Beli and James M. Dominguez II contributed equally to this work.

    An erratum to this article is available at http://dx.doi.org/10.1007/s00109-017-1530-8.

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    ESM 1

    Supplemental Figure S1: Gating strategy for single cell suspension of brain and retina. Viable/Live singlets were gated on, CD45+, CD45hi recruits, and then F4/80+/- Ly6G- mononuclear cells. Using this gating strategy allowed subsequent analysis of Ly6C+ monocytes and Ly6C- monocyte/macrophage. Data shown are from brain and retina of STZ-diabetic WT mice. Supplemental Figure S2: Gating strategy for single cell suspension of bone marrow and spleen. Splenic and bone marrow monocytes were gated in the lymphocyte gate based on forward and side scatter and identified as Lineage 1 (CD3e, CD19, NK1.1, Ly6G) negative, Lineage 2 (CD11c, F4/80, IA-b) negative and CD11b+ cells (PPTX 379 kb)

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    Beli, E., Dominguez, J.M., Hu, P. et al. CX3CR1 deficiency accelerates the development of retinopathy in a rodent model of type 1 diabetes. J Mol Med 94, 1255–1265 (2016). https://doi.org/10.1007/s00109-016-1433-0

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