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Complement component 3 from astrocytes mediates retinal ganglion cell loss during neuroinflammation

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Abstract

Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS) characterized by varying degrees of secondary neurodegeneration. Retinal ganglion cells (RGC) are lost in MS in association with optic neuritis but the mechanisms of neuronal injury remain unclear. Complement component C3 has been implicated in retinal and cerebral synaptic pathology that may precede neurodegeneration. Herein, we examined post-mortem MS retinas, and then used a mouse model, experimental autoimmune encephalomyelitis (EAE), to examine the role of C3 in the pathogenesis of RGC loss associated with optic neuritis. First, we show extensive C3 expression in astrocytes (C3+/GFAP+ cells) and significant RGC loss (RBPMS+ cells) in post-mortem retinas from people with MS compared to retinas from non-MS individuals. A patient with progressive MS with a remote history of optic neuritis showed marked reactive astrogliosis with C3 expression in the inner retina extending into deeper layers in the affected eye more than the unaffected eye. To study whether C3 mediates retinal degeneration, we utilized global C3–/– EAE mice and found that they had less RGC loss and partially preserved neurites in the retina compared with C3+/+ EAE mice. C3–/– EAE mice had fewer axonal swellings in the optic nerve, reflecting reduced axonal injury, but had no changes in demyelination or T cell infiltration into the CNS. Using a C3-tdTomato reporter mouse line, we show definitive evidence of C3 expression in astrocytes in the retina and optic nerves of EAE mice. Conditional deletion of C3 in astrocytes showed RGC protection replicating the effects seen in the global knockouts. These data implicate astrocyte C3 expression as a critical mediator of retinal neuronal pathology in EAE and MS, and are consistent with recent studies showing C3 gene variants are associated with faster rates of retinal neurodegeneration in human disease.

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Acknowledgements

The authors wish to thank Kyle Martin, Arthur Anthony Reyes, and Hannah-Noelle Lord for assistance with scoring EAE mice. The authors also thank the JHU Ross Flow Cytometry Core for use of a Cytek Aurora with a yellow laser to assess tdT expression.

Funding

This study was funded by; R01 NS041435 (PAC), National MS Society (NMSS) grants RG-1907–34756 (PAC) and SI-2004–36590 (DSR), the Division of Intramural Research of the National Institute of Neurological Disorders and Stroke (NINDS) and the National Heart, Lung, and Blood Institute (NHLBI), NIH (ZIANS003119 to DSR and ZIAHL006223 to CK), Fonds de recherche du Québec—Santé (FRQS)# 270746 to MG, and NIH S10OD026859 to JHU.

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  1. Marjan Gharagozloo, Matthew D. Smith and Jing Jin contributed equally.

Authors and Affiliations

  1. Division of Neuroimmunology and Neurological Infections, Johns Hopkins Hospital, Pathology Building 509, 600 N. Wolfe St., Baltimore, Md, 21287, USA

    Marjan Gharagozloo, Matthew D. Smith, Jing Jin, Thomas Garton, Michelle Taylor, Alyssa Chao, Keya Meyers, Michael D. Kornberg, Daniel S. Reich, Katharine A. Whartenby & Peter A. Calabresi

  2. Solomon H. Snyder Department of Neuroscience, Johns Hopkins University, Baltimore, MD, 21205, USA

    Donald J. Zack & Peter A. Calabresi

  3. Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Donald J. Zack, Charles G. Eberhart & Peter A. Calabresi

  4. The Guerrieri Center for Genetic Engineering and Molecular Ophthalmology, The Wilmer Eye Institute, The Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    Donald J. Zack

  5. Neuroimmunology Clinic and Translational Neuroradiology Section National Institute of Neurological Disorders and Stroke, Bethesda, MD, 20892, USA

    Joan Ohayon, Brent A. Calabresi & Daniel S. Reich

  6. Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA

    Charles G. Eberhart

  7. Complement and Inflammation Research Section (CIRS), National Heart, Lung and Blood Institute, Bethesda, MD, 20892, USA

    Claudia Kemper

  8. Institute for Systemic Inflammation Research, University of Lübeck, 23562, Lübeck, Germany

    Claudia Kemper

  9. Departments of Neurology and Pathology, Johns Hopkins Hospital, Pathology Building 509, 600 N. Wolfe St., Baltimore, Md, 21287, USA

    Carlos A. Pardo

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  1. Marjan Gharagozloo
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Gharagozloo, M., Smith, M.D., Jin, J. et al. Complement component 3 from astrocytes mediates retinal ganglion cell loss during neuroinflammation. Acta Neuropathol 142, 899–915 (2021). https://doi.org/10.1007/s00401-021-02366-4

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