Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease

Sebastian Werneburg; Jonathan Jung; Rejani B. Kunjamma; Seung Kwon Ha; Nicholas J. Luciano; Cory M. Willis; Guangping Gao; Natalia P. Biscola; Leif A. Havton; Stephen J. Crocker; Brian Popko; Daniel S. Reich; Dorothy P. Schafer

doi:10.1016/j.immuni.2019.12.004

Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease

Sebastian Werneburg, Jonathan Jung, Rejani B. Kunjamma, Seung Kwon Ha, Nicholas J. Luciano, Cory M. Willis, Guangping Gao, Natalia P. Biscola, Leif A. Havton, Stephen J. Crocker, Brian Popko, Daniel S. Reich, Dorothy P. Schafer^*

^*Corresponding author for this work

Neurology

Research output: Contribution to journal › Article › peer-review

258 Scopus citations

Abstract

Multiple sclerosis (MS) is a demyelinating, autoimmune disease of the central nervous system. While work has focused on myelin and axon loss in MS, less is known about mechanisms underlying synaptic changes. Using postmortem human MS tissue, a preclinical nonhuman primate model of MS, and two rodent models of demyelinating disease, we investigated synapse changes in the visual system. Similar to other neurodegenerative diseases, microglial synaptic engulfment and profound synapse loss were observed. In mice, synapse loss occurred independently of local demyelination and neuronal degeneration but coincided with gliosis and increased complement component C3, but not C1q, at synapses. Viral overexpression of the complement inhibitor Crry at C3-bound synapses decreased microglial engulfment of synapses and protected visual function. These results indicate that microglia eliminate synapses through the alternative complement cascade in demyelinating disease and identify a strategy to prevent synapse loss that may be broadly applicable to other neurodegenerative diseases. Video Abstract: The mechanisms underlying synaptic changes in multiple sclerosis (MS) remain unclear. Werneburg et al. identify microglia-mediated synapse engulfment and synapse loss in MS patients and multiple MS-relevant animal models. Synapse loss can occur early and prior to other MS-relevant pathology and is associated with synapse-localized complement C3. An AAV approach to inhibit C3 protects synapses and preserves circuit function.

Original language	English (US)
Pages (from-to)	167-182.e7
Journal	Immunity
Volume	52
Issue number	1
DOIs	https://doi.org/10.1016/j.immuni.2019.12.004
State	Published - Jan 14 2020

Funding

We thank Dr. Christopher C. Hemond (UMMS) for critical reading of the manuscript. We thank Dr. Paola Perrat (UMMS) for advice on the design and production of plasmids, Shannon Becker (UMMS) and Anoushka Lotun (UMMS) for assistance with tissue preparation, and Dr. Claudio Punzo and Georgia Gunner for advice and training on optomotor testing. We further thank Prof. Dr. Matthew Rasband (Baylor College of Medicine) for sharing CASPR and βIV-spectrin specific antibodies and Prof. Oleg Butovsky for providing P2RY12 specific antibodies. This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany) grant WE 6170/1-1 (S.W.), NIMH— RO1MH113743 (D.P.S.), the Intramural Research Program of NINDS (D.S.R.), NINDS— R01NS099334 (B.P.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (D.P.S., D.S.R., B.P., L.A.H.). We thank Dr. Christopher C. Hemond (UMMS) for critical reading of the manuscript. We thank Dr. Paola Perrat (UMMS) for advice on the design and production of plasmids, Shannon Becker (UMMS) and Anoushka Lotun (UMMS) for assistance with tissue preparation, and Dr. Claudio Punzo and Georgia Gunner for advice and training on optomotor testing. We further thank Prof. Dr. Matthew Rasband (Baylor College of Medicine) for sharing CASPR and ?IV-spectrin specific antibodies and Prof. Oleg Butovsky for providing P2RY12 specific antibodies. This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany) grant WE 6170/1-1 (S.W.), NIMH?RO1MH113743 (D.P.S.), the Intramural Research Program of NINDS (D.S.R.), NINDS?R01NS099334 (B.P.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (D.P.S. D.S.R. B.P. L.A.H.). S.W. and D.P.S. designed the study and wrote the manuscript. S.W. performed and analyzed the experiments. J.J. assisted with mouse maintenance and conduction and analysis of experiments. S.J.C. and C.M.W. provided first murine EAE tissue and assisted in establishing the EAE mouse model. G.G. assisted with viral vector production. B.P. and R.B.K. maintained DTA mice and collected tissue for analyses. D.S.R. supervised collection and processing of human brain tissue by S.-K.H. and treatment and collection of marmoset tissue by N.J.L. N.P.B. and L.A.H. assisted with electron microscopy studies and measurements of G-ratios. All authors revised the manuscript for intellectually important content. The authors declare no conflict of interest.

Keywords

complement
demyelination
engulfment
gene therapy
microglia
multiple sclerosis
neural-immune
neurodegeneration
neuroinflammation
synapse

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.immuni.2019.12.004

Cite this

Werneburg, S., Jung, J., Kunjamma, R. B., Ha, S. K., Luciano, N. J., Willis, C. M., Gao, G., Biscola, N. P., Havton, L. A., Crocker, S. J., Popko, B., Reich, D. S., & Schafer, D. P. (2020). Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease. Immunity, 52(1), 167-182.e7. https://doi.org/10.1016/j.immuni.2019.12.004

@article{df88a2ad7af54eb4b453b0ec12da3de7,

title = "Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease",

abstract = "Multiple sclerosis (MS) is a demyelinating, autoimmune disease of the central nervous system. While work has focused on myelin and axon loss in MS, less is known about mechanisms underlying synaptic changes. Using postmortem human MS tissue, a preclinical nonhuman primate model of MS, and two rodent models of demyelinating disease, we investigated synapse changes in the visual system. Similar to other neurodegenerative diseases, microglial synaptic engulfment and profound synapse loss were observed. In mice, synapse loss occurred independently of local demyelination and neuronal degeneration but coincided with gliosis and increased complement component C3, but not C1q, at synapses. Viral overexpression of the complement inhibitor Crry at C3-bound synapses decreased microglial engulfment of synapses and protected visual function. These results indicate that microglia eliminate synapses through the alternative complement cascade in demyelinating disease and identify a strategy to prevent synapse loss that may be broadly applicable to other neurodegenerative diseases. Video Abstract: The mechanisms underlying synaptic changes in multiple sclerosis (MS) remain unclear. Werneburg et al. identify microglia-mediated synapse engulfment and synapse loss in MS patients and multiple MS-relevant animal models. Synapse loss can occur early and prior to other MS-relevant pathology and is associated with synapse-localized complement C3. An AAV approach to inhibit C3 protects synapses and preserves circuit function.",

keywords = "complement, demyelination, engulfment, gene therapy, microglia, multiple sclerosis, neural-immune, neurodegeneration, neuroinflammation, synapse",

author = "Sebastian Werneburg and Jonathan Jung and Kunjamma, {Rejani B.} and Ha, {Seung Kwon} and Luciano, {Nicholas J.} and Willis, {Cory M.} and Guangping Gao and Biscola, {Natalia P.} and Havton, {Leif A.} and Crocker, {Stephen J.} and Brian Popko and Reich, {Daniel S.} and Schafer, {Dorothy P.}",

note = "Funding Information: We thank Dr. Christopher C. Hemond (UMMS) for critical reading of the manuscript. We thank Dr. Paola Perrat (UMMS) for advice on the design and production of plasmids, Shannon Becker (UMMS) and Anoushka Lotun (UMMS) for assistance with tissue preparation, and Dr. Claudio Punzo and Georgia Gunner for advice and training on optomotor testing. We further thank Prof. Dr. Matthew Rasband (Baylor College of Medicine) for sharing CASPR and βIV-spectrin specific antibodies and Prof. Oleg Butovsky for providing P2RY12 specific antibodies. This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany) grant WE 6170/1-1 (S.W.), NIMH— RO1MH113743 (D.P.S.), the Intramural Research Program of NINDS (D.S.R.), NINDS— R01NS099334 (B.P.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (D.P.S., D.S.R., B.P., L.A.H.). Funding Information: We thank Dr. Christopher C. Hemond (UMMS) for critical reading of the manuscript. We thank Dr. Paola Perrat (UMMS) for advice on the design and production of plasmids, Shannon Becker (UMMS) and Anoushka Lotun (UMMS) for assistance with tissue preparation, and Dr. Claudio Punzo and Georgia Gunner for advice and training on optomotor testing. We further thank Prof. Dr. Matthew Rasband (Baylor College of Medicine) for sharing CASPR and ?IV-spectrin specific antibodies and Prof. Oleg Butovsky for providing P2RY12 specific antibodies. This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany) grant WE 6170/1-1 (S.W.), NIMH?RO1MH113743 (D.P.S.), the Intramural Research Program of NINDS (D.S.R.), NINDS?R01NS099334 (B.P.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (D.P.S. D.S.R. B.P. L.A.H.). S.W. and D.P.S. designed the study and wrote the manuscript. S.W. performed and analyzed the experiments. J.J. assisted with mouse maintenance and conduction and analysis of experiments. S.J.C. and C.M.W. provided first murine EAE tissue and assisted in establishing the EAE mouse model. G.G. assisted with viral vector production. B.P. and R.B.K. maintained DTA mice and collected tissue for analyses. D.S.R. supervised collection and processing of human brain tissue by S.-K.H. and treatment and collection of marmoset tissue by N.J.L. N.P.B. and L.A.H. assisted with electron microscopy studies and measurements of G-ratios. All authors revised the manuscript for intellectually important content. The authors declare no conflict of interest. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",

year = "2020",

month = jan,

day = "14",

doi = "10.1016/j.immuni.2019.12.004",

language = "English (US)",

volume = "52",

pages = "167--182.e7",

journal = "Immunity",

issn = "1074-7613",

publisher = "Cell Press",

number = "1",

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Werneburg, S, Jung, J, Kunjamma, RB, Ha, SK, Luciano, NJ, Willis, CM, Gao, G, Biscola, NP, Havton, LA, Crocker, SJ, Popko, B, Reich, DS & Schafer, DP 2020, 'Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease', Immunity, vol. 52, no. 1, pp. 167-182.e7. https://doi.org/10.1016/j.immuni.2019.12.004

TY - JOUR

T1 - Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease

AU - Werneburg, Sebastian

AU - Jung, Jonathan

AU - Kunjamma, Rejani B.

AU - Ha, Seung Kwon

AU - Luciano, Nicholas J.

AU - Willis, Cory M.

AU - Gao, Guangping

AU - Biscola, Natalia P.

AU - Havton, Leif A.

AU - Crocker, Stephen J.

AU - Popko, Brian

AU - Reich, Daniel S.

AU - Schafer, Dorothy P.

N1 - Funding Information: We thank Dr. Christopher C. Hemond (UMMS) for critical reading of the manuscript. We thank Dr. Paola Perrat (UMMS) for advice on the design and production of plasmids, Shannon Becker (UMMS) and Anoushka Lotun (UMMS) for assistance with tissue preparation, and Dr. Claudio Punzo and Georgia Gunner for advice and training on optomotor testing. We further thank Prof. Dr. Matthew Rasband (Baylor College of Medicine) for sharing CASPR and βIV-spectrin specific antibodies and Prof. Oleg Butovsky for providing P2RY12 specific antibodies. This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany) grant WE 6170/1-1 (S.W.), NIMH— RO1MH113743 (D.P.S.), the Intramural Research Program of NINDS (D.S.R.), NINDS— R01NS099334 (B.P.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (D.P.S., D.S.R., B.P., L.A.H.). Funding Information: We thank Dr. Christopher C. Hemond (UMMS) for critical reading of the manuscript. We thank Dr. Paola Perrat (UMMS) for advice on the design and production of plasmids, Shannon Becker (UMMS) and Anoushka Lotun (UMMS) for assistance with tissue preparation, and Dr. Claudio Punzo and Georgia Gunner for advice and training on optomotor testing. We further thank Prof. Dr. Matthew Rasband (Baylor College of Medicine) for sharing CASPR and ?IV-spectrin specific antibodies and Prof. Oleg Butovsky for providing P2RY12 specific antibodies. This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany) grant WE 6170/1-1 (S.W.), NIMH?RO1MH113743 (D.P.S.), the Intramural Research Program of NINDS (D.S.R.), NINDS?R01NS099334 (B.P.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (D.P.S. D.S.R. B.P. L.A.H.). S.W. and D.P.S. designed the study and wrote the manuscript. S.W. performed and analyzed the experiments. J.J. assisted with mouse maintenance and conduction and analysis of experiments. S.J.C. and C.M.W. provided first murine EAE tissue and assisted in establishing the EAE mouse model. G.G. assisted with viral vector production. B.P. and R.B.K. maintained DTA mice and collected tissue for analyses. D.S.R. supervised collection and processing of human brain tissue by S.-K.H. and treatment and collection of marmoset tissue by N.J.L. N.P.B. and L.A.H. assisted with electron microscopy studies and measurements of G-ratios. All authors revised the manuscript for intellectually important content. The authors declare no conflict of interest. Publisher Copyright: © 2019 Elsevier Inc.

PY - 2020/1/14

Y1 - 2020/1/14

N2 - Multiple sclerosis (MS) is a demyelinating, autoimmune disease of the central nervous system. While work has focused on myelin and axon loss in MS, less is known about mechanisms underlying synaptic changes. Using postmortem human MS tissue, a preclinical nonhuman primate model of MS, and two rodent models of demyelinating disease, we investigated synapse changes in the visual system. Similar to other neurodegenerative diseases, microglial synaptic engulfment and profound synapse loss were observed. In mice, synapse loss occurred independently of local demyelination and neuronal degeneration but coincided with gliosis and increased complement component C3, but not C1q, at synapses. Viral overexpression of the complement inhibitor Crry at C3-bound synapses decreased microglial engulfment of synapses and protected visual function. These results indicate that microglia eliminate synapses through the alternative complement cascade in demyelinating disease and identify a strategy to prevent synapse loss that may be broadly applicable to other neurodegenerative diseases. Video Abstract: The mechanisms underlying synaptic changes in multiple sclerosis (MS) remain unclear. Werneburg et al. identify microglia-mediated synapse engulfment and synapse loss in MS patients and multiple MS-relevant animal models. Synapse loss can occur early and prior to other MS-relevant pathology and is associated with synapse-localized complement C3. An AAV approach to inhibit C3 protects synapses and preserves circuit function.

AB - Multiple sclerosis (MS) is a demyelinating, autoimmune disease of the central nervous system. While work has focused on myelin and axon loss in MS, less is known about mechanisms underlying synaptic changes. Using postmortem human MS tissue, a preclinical nonhuman primate model of MS, and two rodent models of demyelinating disease, we investigated synapse changes in the visual system. Similar to other neurodegenerative diseases, microglial synaptic engulfment and profound synapse loss were observed. In mice, synapse loss occurred independently of local demyelination and neuronal degeneration but coincided with gliosis and increased complement component C3, but not C1q, at synapses. Viral overexpression of the complement inhibitor Crry at C3-bound synapses decreased microglial engulfment of synapses and protected visual function. These results indicate that microglia eliminate synapses through the alternative complement cascade in demyelinating disease and identify a strategy to prevent synapse loss that may be broadly applicable to other neurodegenerative diseases. Video Abstract: The mechanisms underlying synaptic changes in multiple sclerosis (MS) remain unclear. Werneburg et al. identify microglia-mediated synapse engulfment and synapse loss in MS patients and multiple MS-relevant animal models. Synapse loss can occur early and prior to other MS-relevant pathology and is associated with synapse-localized complement C3. An AAV approach to inhibit C3 protects synapses and preserves circuit function.

KW - complement

KW - demyelination

KW - engulfment

KW - gene therapy

KW - microglia

KW - multiple sclerosis

KW - neural-immune

KW - neurodegeneration

KW - neuroinflammation

KW - synapse

UR - http://www.scopus.com/inward/record.url?scp=85077636150&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85077636150&partnerID=8YFLogxK

U2 - 10.1016/j.immuni.2019.12.004

DO - 10.1016/j.immuni.2019.12.004

M3 - Article

C2 - 31883839

AN - SCOPUS:85077636150

SN - 1074-7613

VL - 52

SP - 167-182.e7

JO - Immunity

JF - Immunity

IS - 1

ER -

Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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