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A flow cytometry-based in vitro assay reveals that formation of apolipoprotein E (ApoE)-amyloid beta complexes depends on ApoE isoform and cell type.
J Biol Chem. 2018 08 24; 293(34):13247-13256.JB

Abstract

Apolipoprotein E (ApoE) is a secreted apolipoprotein with three isoforms, E2, E3, and E4, that binds to lipids and facilitates their transport in the extracellular environment of the brain and the periphery. The E4 allele is a major genetic risk factor for the sporadic form of Alzheimer's disease (AD), and studies of human brain and mouse models have revealed that E4 significantly exacerbates the deposition of amyloid beta (Aβ). It has been suggested that this deposition could be attributed to the formation of soluble ApoE isoform-specific ApoE-Aβ complexes. However, previous studies have reported conflicting results regarding the directionality and strength of those interactions. In this study, using a series of flow cytometry assays that maintain the physiological integrity of ApoE-Aβ complexes, we systematically assessed the association of Aβ with ApoE2, E3, or E4. We used ApoE secreted from HEK cells or astrocytes overexpressing ApoE fused with a GFP tag. As a source of soluble Aβ peptide, we used synthetic Aβ40 or Aβ42 or physiological Aβ secreted from CHO cell lines overexpressing WT or V717F variant amyloid precursor protein (APP). We observed significant interactions between the different ApoE isoforms and Aβ, with E4 interacting with Aβ more strongly than the E2 and E3 isoforms. We also found subtle differences depending on the Aβ type and the ApoE-producing cell type. In conclusion, these results indicate that the strength of the ApoE-Aβ association depends on the source of Aβ or ApoE.

Authors+Show Affiliations

From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129.From the Alzheimer's Disease Research Laboratory, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129 BHYMAN@mgh.harvard.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29950521

Citation

Kara, Eleanna, et al. "A Flow Cytometry-based in Vitro Assay Reveals That Formation of Apolipoprotein E (ApoE)-amyloid Beta Complexes Depends On ApoE Isoform and Cell Type." The Journal of Biological Chemistry, vol. 293, no. 34, 2018, pp. 13247-13256.
Kara E, Marks JD, Roe AD, et al. A flow cytometry-based in vitro assay reveals that formation of apolipoprotein E (ApoE)-amyloid beta complexes depends on ApoE isoform and cell type. J Biol Chem. 2018;293(34):13247-13256.
Kara, E., Marks, J. D., Roe, A. D., Commins, C., Fan, Z., Calvo-Rodriguez, M., Wegmann, S., Hudry, E., & Hyman, B. T. (2018). A flow cytometry-based in vitro assay reveals that formation of apolipoprotein E (ApoE)-amyloid beta complexes depends on ApoE isoform and cell type. The Journal of Biological Chemistry, 293(34), 13247-13256. https://doi.org/10.1074/jbc.RA117.001388
Kara E, et al. A Flow Cytometry-based in Vitro Assay Reveals That Formation of Apolipoprotein E (ApoE)-amyloid Beta Complexes Depends On ApoE Isoform and Cell Type. J Biol Chem. 2018 08 24;293(34):13247-13256. PubMed PMID: 29950521.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A flow cytometry-based in vitro assay reveals that formation of apolipoprotein E (ApoE)-amyloid beta complexes depends on ApoE isoform and cell type. AU - Kara,Eleanna, AU - Marks,Jordan D, AU - Roe,Allyson D, AU - Commins,Caitlin, AU - Fan,Zhanyun, AU - Calvo-Rodriguez,Maria, AU - Wegmann,Susanne, AU - Hudry,Eloise, AU - Hyman,Bradley T, Y1 - 2018/06/27/ PY - 2017/12/09/received PY - 2018/05/21/revised PY - 2018/6/29/pubmed PY - 2019/3/1/medline PY - 2018/6/29/entrez KW - Alzheimer disease KW - amyloid beta (AB) KW - apolipoprotein E (ApoE) KW - astrocyte KW - flow cytometry SP - 13247 EP - 13256 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 293 IS - 34 N2 - Apolipoprotein E (ApoE) is a secreted apolipoprotein with three isoforms, E2, E3, and E4, that binds to lipids and facilitates their transport in the extracellular environment of the brain and the periphery. The E4 allele is a major genetic risk factor for the sporadic form of Alzheimer's disease (AD), and studies of human brain and mouse models have revealed that E4 significantly exacerbates the deposition of amyloid beta (Aβ). It has been suggested that this deposition could be attributed to the formation of soluble ApoE isoform-specific ApoE-Aβ complexes. However, previous studies have reported conflicting results regarding the directionality and strength of those interactions. In this study, using a series of flow cytometry assays that maintain the physiological integrity of ApoE-Aβ complexes, we systematically assessed the association of Aβ with ApoE2, E3, or E4. We used ApoE secreted from HEK cells or astrocytes overexpressing ApoE fused with a GFP tag. As a source of soluble Aβ peptide, we used synthetic Aβ40 or Aβ42 or physiological Aβ secreted from CHO cell lines overexpressing WT or V717F variant amyloid precursor protein (APP). We observed significant interactions between the different ApoE isoforms and Aβ, with E4 interacting with Aβ more strongly than the E2 and E3 isoforms. We also found subtle differences depending on the Aβ type and the ApoE-producing cell type. In conclusion, these results indicate that the strength of the ApoE-Aβ association depends on the source of Aβ or ApoE. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/29950521/A_flow_cytometry_based_in_vitro_assay_reveals_that_formation_of_apolipoprotein_E__ApoE__amyloid_beta_complexes_depends_on_ApoE_isoform_and_cell_type_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=29950521 DB - PRIME DP - Unbound Medicine ER -