Antibody blood-brain barrier efflux is modulated by glycan modification

Drug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain penetration of antibody drugs improves Alzheimer's treatment outcomes. New approaches are needed to better accomplish this goal. Based on pr...

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Published inBiochimica et biophysica acta Vol. 1861; no. 9; pp. 2228 - 2239
Main Authors Finke, John M., Ayres, Kari R., Brisbin, Ryan P., Hill, Hali A., Wing, Emily E., Banks, William A.
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.09.2017
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Abstract Drug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain penetration of antibody drugs improves Alzheimer's treatment outcomes. New approaches are needed to better accomplish this goal. Based on prior evidence, the hypothesis that glycan modification alters antibody blood-brain barrier permeability was tested here. The blood-brain barrier permeability coefficient Pe of different glycosylated states of anti-amyloid IgG was measured using in vitro models of brain microvascular endothelial cells. Monoclonal antibodies 4G8, with sialic acid, and 6E10, lacking sialic acid, were studied. The amount of sialic acid was determined using quantitative and semi-quantitative surface plasmon resonance methods. Influx of IgG was not saturable and was largely insensitive to IgG species and glycosylation state. By contrast, efflux of 4G8 efflux was significantly lower than both albumin controls and 6E10. Removal of α2,6-linked sialic acid group present on 12% of 4G8 completely restored efflux to that of 6E10 but increasing the α2,6-sialylated fraction to 15% resulted in no change. Removal of the Fc glycan from 4G8 partially restored efflux. Alternate sialic acid groups with α2,3 and α2,8 linkages, nor on the Fc glycan, were not detected at significant levels on either 4G8 or 6E10. These results support a model in which surface-sialylated 4G8 inhibits its own efflux and that of asialylated 4G8. Glycan modification has the potential to increase antibody drug penetration into the brain through efflux inhibition. •IgG glycan sialic acid inhibits IgG efflux, but not influx, through the blood-brain barrier.•The Fc glycan is required for efflux inhibition by sialylated IgG but does not affect asialylated IgG efflux.•Fab α2,6 sialylation in 12% of IgG inhibits efflux of the asialyalted IgG majority.
AbstractList Drug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain penetration of antibody drugs improves Alzheimer's treatment outcomes. New approaches are needed to better accomplish this goal. Based on prior evidence, the hypothesis that glycan modification alters antibody blood-brain barrier permeability was tested here. The blood-brain barrier permeability coefficient Pe of different glycosylated states of anti-amyloid IgG was measured using in vitro models of brain microvascular endothelial cells. Monoclonal antibodies 4G8, with sialic acid, and 6E10, lacking sialic acid, were studied. The amount of sialic acid was determined using quantitative and semi-quantitative surface plasmon resonance methods. Influx of IgG was not saturable and was largely insensitive to IgG species and glycosylation state. By contrast, efflux of 4G8 efflux was significantly lower than both albumin controls and 6E10. Removal of α2,6-linked sialic acid group present on 12% of 4G8 completely restored efflux to that of 6E10 but increasing the α2,6-sialylated fraction to 15% resulted in no change. Removal of the Fc glycan from 4G8 partially restored efflux. Alternate sialic acid groups with α2,3 and α2,8 linkages, nor on the Fc glycan, were not detected at significant levels on either 4G8 or 6E10. These results support a model in which surface-sialylated 4G8 inhibits its own efflux and that of asialylated 4G8. Glycan modification has the potential to increase antibody drug penetration into the brain through efflux inhibition. •IgG glycan sialic acid inhibits IgG efflux, but not influx, through the blood-brain barrier.•The Fc glycan is required for efflux inhibition by sialylated IgG but does not affect asialylated IgG efflux.•Fab α2,6 sialylation in 12% of IgG inhibits efflux of the asialyalted IgG majority.
BACKGROUNDDrug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain penetration of antibody drugs improves Alzheimer's treatment outcomes. New approaches are needed to better accomplish this goal. Based on prior evidence, the hypothesis that glycan modification alters antibody blood-brain barrier permeability was tested here.METHODSThe blood-brain barrier permeability coefficient Pe of different glycosylated states of anti-amyloid IgG was measured using in vitro models of brain microvascular endothelial cells. Monoclonal antibodies 4G8, with sialic acid, and 6E10, lacking sialic acid, were studied. The amount of sialic acid was determined using quantitative and semi-quantitative surface plasmon resonance methods.RESULTSInflux of IgG was not saturable and was largely insensitive to IgG species and glycosylation state. By contrast, efflux of 4G8 efflux was significantly lower than both albumin controls and 6E10. Removal of α2,6-linked sialic acid group present on 12% of 4G8 completely restored efflux to that of 6E10 but increasing the α2,6-sialylated fraction to 15% resulted in no change. Removal of the Fc glycan from 4G8 partially restored efflux. Alternate sialic acid groups with α2,3 and α2,8 linkages, nor on the Fc glycan, were not detected at significant levels on either 4G8 or 6E10.CONCLUSIONSThese results support a model in which surface-sialylated 4G8 inhibits its own efflux and that of asialylated 4G8.GENERAL SIGNIFICANCEGlycan modification has the potential to increase antibody drug penetration into the brain through efflux inhibition.
Drug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain penetration of antibody drugs improves Alzheimer's treatment outcomes. New approaches are needed to better accomplish this goal. Based on prior evidence, the hypothesis that glycan modification alters antibody blood-brain barrier permeability was tested here. The blood-brain barrier permeability coefficient Pe of different glycosylated states of anti-amyloid IgG was measured using in vitro models of brain microvascular endothelial cells. Monoclonal antibodies 4G8, with sialic acid, and 6E10, lacking sialic acid, were studied. The amount of sialic acid was determined using quantitative and semi-quantitative surface plasmon resonance methods. Influx of IgG was not saturable and was largely insensitive to IgG species and glycosylation state. By contrast, efflux of 4G8 efflux was significantly lower than both albumin controls and 6E10. Removal of α2,6-linked sialic acid group present on 12% of 4G8 completely restored efflux to that of 6E10 but increasing the α2,6-sialylated fraction to 15% resulted in no change. Removal of the Fc glycan from 4G8 partially restored efflux. Alternate sialic acid groups with α2,3 and α2,8 linkages, nor on the Fc glycan, were not detected at significant levels on either 4G8 or 6E10. These results support a model in which surface-sialylated 4G8 inhibits its own efflux and that of asialylated 4G8. Glycan modification has the potential to increase antibody drug penetration into the brain through efflux inhibition.
Author Ayres, Kari R.
Wing, Emily E.
Brisbin, Ryan P.
Hill, Hali A.
Finke, John M.
Banks, William A.
AuthorAffiliation b Geriatric Research Education and Clinical Center, VA Puget Sound Health Care System, Seattle, WA, USA
a Division of Sciences and Mathematics, Interdisciplinary Arts and Sciences, University of Washington Tacoma, Tacoma, WA, USA
c Division of Gerontology and Geriatric Medicine, Department of Geriatric Medicine, University of Washington School of Medicine, Seattle, WA, USA
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Issue 9
Keywords Fab Fc antibody glycosylation
Sialic acid
Surface plasmon resonance
Alzheimer's disease
Blood brain barrier
Language English
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Snippet Drug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain...
BACKGROUNDDrug delivery to the brain is a major roadblock to treatment of Alzheimer's disease. Recent results of the PRIME study indicate that increasing brain...
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StartPage 2228
SubjectTerms Alzheimer's disease
Animals
Antibodies, Monoclonal - analysis
Antibodies, Monoclonal - metabolism
Blood brain barrier
Fab Fc antibody glycosylation
Glycosylation
Immunoglobulin G - analysis
Immunoglobulin G - metabolism
Male
Mice
N-Acetylneuraminic Acid - analysis
Sialic acid
Surface plasmon resonance
Title Antibody blood-brain barrier efflux is modulated by glycan modification
URI https://dx.doi.org/10.1016/j.bbagen.2017.06.008
https://www.ncbi.nlm.nih.gov/pubmed/28624473
https://search.proquest.com/docview/1911198673
https://pubmed.ncbi.nlm.nih.gov/PMC5557653
Volume 1861
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