Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease

[Background] Astrocytes and microglia react to Aβ plaques, neurofibrillary tangles, and neurodegeneration in the Alzheimer's disease (AD) brain. Single-nuclei and single-cell RNA-seq have revealed multiple states or subpopulations of these glial cells but lack spatial information. We have devel...

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Authors: Muñoz-Castro, Clara, Noori, Ayush, Magdamo, Colin G., Li, Zhaozhi, Marks, Jordan D., Frosch, Matthew P., Das, Sudeshna, Hyman, Bradley T., Serrano-Pozo, Alberto
Format: article
Status:Published version
Publication Date:2022
Country:España
Institution:Consejo Superior de Investigaciones Científicas (CSIC)
Repository:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/305309
Online Access:http://hdl.handle.net/10261/305309
https://api.elsevier.com/content/abstract/scopus_id/85123973861
Access Level:Open access
Keyword:Alzheimer’s disease
Amyloid plaques
Astrocytes
Immunohistochemistry
Microglia
Neurofibrillary tangles
Neuropathology
Tau
id ES_0bfa93684b3f437fc9225e71c8bf8d9c
oai_identifier_str oai:digital.csic.es:10261/305309
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
title Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
spellingShingle Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
Muñoz-Castro, Clara
Alzheimer’s disease
Amyloid plaques
Astrocytes
Immunohistochemistry
Microglia
Neurofibrillary tangles
Neuropathology
Tau
title_short Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
title_full Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
title_fullStr Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
title_full_unstemmed Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
title_sort Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's disease
dc.creator.none.fl_str_mv Muñoz-Castro, Clara
Noori, Ayush
Magdamo, Colin G.
Li, Zhaozhi
Marks, Jordan D.
Frosch, Matthew P.
Das, Sudeshna
Hyman, Bradley T.
Serrano-Pozo, Alberto
author Muñoz-Castro, Clara
author_facet Muñoz-Castro, Clara
Noori, Ayush
Magdamo, Colin G.
Li, Zhaozhi
Marks, Jordan D.
Frosch, Matthew P.
Das, Sudeshna
Hyman, Bradley T.
Serrano-Pozo, Alberto
author_role author
author2 Noori, Ayush
Magdamo, Colin G.
Li, Zhaozhi
Marks, Jordan D.
Frosch, Matthew P.
Das, Sudeshna
Hyman, Bradley T.
Serrano-Pozo, Alberto
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Agencia Estatal de Investigación (España)
Harvard University
National Institute on Aging (US)
Massachusetts Alzheimer's Disease Research Center
Alzheimer's Association
Serrano-Pozo, Alberto [0000-0003-0899-7530]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Alzheimer’s disease
Amyloid plaques
Astrocytes
Immunohistochemistry
Microglia
Neurofibrillary tangles
Neuropathology
Tau
topic Alzheimer’s disease
Amyloid plaques
Astrocytes
Immunohistochemistry
Microglia
Neurofibrillary tangles
Neuropathology
Tau
description [Background] Astrocytes and microglia react to Aβ plaques, neurofibrillary tangles, and neurodegeneration in the Alzheimer's disease (AD) brain. Single-nuclei and single-cell RNA-seq have revealed multiple states or subpopulations of these glial cells but lack spatial information. We have developed a methodology of cyclic multiplex fluorescent immunohistochemistry on human postmortem brains and image analysis that enables a comprehensive morphological quantitative characterization of astrocytes and microglia in the context of their spatial relationships with plaques and tangles.
publishDate 2022
dc.date.none.fl_str_mv 2022
2023
2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/305309
https://api.elsevier.com/content/abstract/scopus_id/85123973861
url http://hdl.handle.net/10261/305309
https://api.elsevier.com/content/abstract/scopus_id/85123973861
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Muñoz-Castro, Clara; Noori, Ayush; Magdamo, Colin G.; Li, Zhaozhi; Marks, Jordan D.; Frosch, Matthew P.; Das, Sudeshna; Hyman, Bradley T.; Serrano-Pozo, Alberto; 2022; Additional file 1 of Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer’s disease [Dataset]; Figshare; https://doi.org/10.6084/m9.figshare.19113054.v1
Muñoz-Castro, Clara; Noori, Ayush; Magdamo, Colin G.; Li, Zhaozhi; Marks, Jordan D.; Frosch, Matthew P.; Das, Sudeshna; Hyman, Bradley T.; Serrano-Pozo, Alberto; 2022; Additional file 2 of Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer’s disease [Dataset]; Figshare; https://doi.org/10.6084/m9.figshare.19113057.v1
https://doi.org/10.1186/s12974-022-02383-4
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer's diseaseMuñoz-Castro, ClaraNoori, AyushMagdamo, Colin G.Li, ZhaozhiMarks, Jordan D.Frosch, Matthew P.Das, SudeshnaHyman, Bradley T.Serrano-Pozo, AlbertoAlzheimer’s diseaseAmyloid plaquesAstrocytesImmunohistochemistryMicrogliaNeurofibrillary tanglesNeuropathologyTau[Background] Astrocytes and microglia react to Aβ plaques, neurofibrillary tangles, and neurodegeneration in the Alzheimer's disease (AD) brain. Single-nuclei and single-cell RNA-seq have revealed multiple states or subpopulations of these glial cells but lack spatial information. We have developed a methodology of cyclic multiplex fluorescent immunohistochemistry on human postmortem brains and image analysis that enables a comprehensive morphological quantitative characterization of astrocytes and microglia in the context of their spatial relationships with plaques and tangles.[Methods] Single FFPE sections from the temporal association cortex of control and AD subjects were subjected to 8 cycles of multiplex fluorescent immunohistochemistry, including 7 astroglial, 6 microglial, 1 neuronal, Aβ, and phospho-tau markers. Our analysis pipeline consisted of: (1) image alignment across cycles; (2) background subtraction; (3) manual annotation of 5172 ALDH1L1+ astrocytic and 6226 IBA1+ microglial profiles; (4) local thresholding and segmentation of profiles; (5) machine learning on marker intensity data; and (6) deep learning on image features.[Results] Spectral clustering identified three phenotypes of astrocytes and microglia, which we termed “homeostatic,” “intermediate,” and “reactive.” Reactive and, to a lesser extent, intermediate astrocytes and microglia were closely associated with AD pathology (≤ 50 µm). Compared to homeostatic, reactive astrocytes contained substantially higher GFAP and YKL-40, modestly elevated vimentin and TSPO as well as EAAT1, and reduced GS. Intermediate astrocytes had markedly increased EAAT2, moderately increased GS, and intermediate GFAP and YKL-40 levels. Relative to homeostatic, reactive microglia showed increased expression of all markers (CD68, ferritin, MHC2, TMEM119, TSPO), whereas intermediate microglia exhibited increased ferritin and TMEM119 as well as intermediate CD68 levels. Machine learning models applied on either high-plex signal intensity data (gradient boosting machines) or directly on image features (convolutional neural networks) accurately discriminated control vs. AD diagnoses at the single-cell level.[Conclusions] Cyclic multiplex fluorescent immunohistochemistry combined with machine learning models holds promise to advance our understanding of the complexity and heterogeneity of glial responses as well as inform transcriptomics studies. Three distinct phenotypes emerged with our combination of markers, thus expanding the classic binary “homeostatic vs. reactive” classification to a third state, which could represent “transitional” or “resilient” glia.This work was supported by the Spanish Ministry of Science, Innovation, and Universities (FPU fellowship to CM-C), the Real Colegio Complutense at Harvard University (Research Fellowship to CM-C), the National Institute on Aging (K08AG064039 to AS-P, NACC New Investigator Award 2019-NI-09 to AS-P, Massachusetts Alzheimer’s Disease Research Center grant P30AG062421 to BTH, and 1R56AG061196 to BTH), and the Alzheimer’s Association (AACF-17-524184 and AACF-17-524184-RAPID to AS-P). The National Alzheimer’s Coordinating Center (NACC) is funded by the National Institute on Aging (U01 AG016976).Peer reviewedBioMed CentralMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de Investigación (España)Harvard UniversityNational Institute on Aging (US)Massachusetts Alzheimer's Disease Research CenterAlzheimer's AssociationSerrano-Pozo, Alberto [0000-0003-0899-7530]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202320232022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/305309https://api.elsevier.com/content/abstract/scopus_id/85123973861reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésMuñoz-Castro, Clara; Noori, Ayush; Magdamo, Colin G.; Li, Zhaozhi; Marks, Jordan D.; Frosch, Matthew P.; Das, Sudeshna; Hyman, Bradley T.; Serrano-Pozo, Alberto; 2022; Additional file 1 of Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer’s disease [Dataset]; Figshare; https://doi.org/10.6084/m9.figshare.19113054.v1Muñoz-Castro, Clara; Noori, Ayush; Magdamo, Colin G.; Li, Zhaozhi; Marks, Jordan D.; Frosch, Matthew P.; Das, Sudeshna; Hyman, Bradley T.; Serrano-Pozo, Alberto; 2022; Additional file 2 of Cyclic multiplex fluorescent immunohistochemistry and machine learning reveal distinct states of astrocytes and microglia in normal aging and Alzheimer’s disease [Dataset]; Figshare; https://doi.org/10.6084/m9.figshare.19113057.v1https://doi.org/10.1186/s12974-022-02383-4Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3053092026-05-22T06:33:51Z
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