CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes

Maria L. Elkjaer; Arkadiusz Nawrocki; Tim Kacprowski; Pernille Lassen; Anja Hviid Simonsen; Romain Marignier; Tobias Sejbaek; Helle H. Nielsen; Lene Wermuth; Alyaa Yakut Rashid; Peter Høgh; Finn Sellebjerg; Richard Reynolds; Jan Baumbach; Martin R. Larsen; Zsolt Illes

doi:10.1038/s41598-021-83591-5

CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes

Maria L. Elkjaer, Arkadiusz Nawrocki, Tim Kacprowski, Pernille Lassen, Anja Hviid Simonsen, Romain Marignier, Tobias Sejbaek, Helle H. Nielsen, Lene Wermuth, Alyaa Yakut Rashid, Peter Høgh, Finn Sellebjerg, Richard Reynolds, Jan Baumbach, Martin R. Larsen, Zsolt Illes^*

^*Kontaktforfatter

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

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Abstrakt

To identify markers in the CSF of multiple sclerosis (MS) subtypes, we used a two-step proteomic approach: (i) Discovery proteomics compared 169 pooled CSF from MS subtypes and inflammatory/degenerative CNS diseases (NMO spectrum and Alzheimer disease) and healthy controls. (ii) Next, 299 proteins selected by comprehensive statistics were quantified in 170 individual CSF samples. (iii) Genes of the identified proteins were also screened among transcripts in 73 MS brain lesions compared to 25 control brains. F-test based feature selection resulted in 8 proteins differentiating the MS subtypes, and secondary progressive (SP)MS was the most different also from controls. Genes of 7 out these 8 proteins were present in MS brain lesions: GOLM was significantly differentially expressed in active, chronic active, inactive and remyelinating lesions, FRZB in active and chronic active lesions, and SELENBP1 in inactive lesions. Volcano maps of normalized proteins in the different disease groups also indicated the highest amount of altered proteins in SPMS. Apolipoprotein C-I, apolipoprotein A-II, augurin, receptor-type tyrosine-protein phosphatase gamma, and trypsin-1 were upregulated in the CSF of MS subtypes compared to controls. This CSF profile and associated brain lesion spectrum highlight non-inflammatory mechanisms in differentiating CNS diseases and MS subtypes and the uniqueness of SPMS.

Originalsprog	Engelsk
Artikelnummer	4132
Tidsskrift	Scientific Reports
Vol/bind	11
Antal sider	13
ISSN	2045-2322
DOI	https://doi.org/10.1038/s41598-021-83591-5
Status	Udgivet - 18. feb. 2021

Bibliografisk note

Funding Information:
Immunohistochemistry and RNAscope of chronic active brain lesion. Human postmortem brain tissue were supplied by the UK Multiple Sclerosis Tissue Bank (UK Multicentre Research Ethics Committee, MREC/02/2/39), funded by the Multiple Sclerosis Society of Great Britain and Northern Ireland (registered charity 207,495). Fresh-frozen blocks containing chronic active lesion from progressive MS patients were sectioned (10-μm), PFA-fixed, blocked in PBS with 10% normal horse serum (NHS) and immunostained with rabbit CHI3L1 (monoclonal antibody) 1:200 (Abcam) followed by biotinylated secondary antibody (Jackson Immu-noresearch Laboratories, Cambridgeshire, UK), avidin/biotin staining (Vector Laboratories, Burlingame, CA) and 3,3′-diaminobenzidine (DAB) staining (Vector Laboratories, Burlingame, CA). The RNAscope 2.5 Duplex Assay (ACD Biosystems) was performed according to the ACD protocol for fresh-frozen tissue. Chronic active lesions were hybridized with two mRNA probes per experiment. Hs-GFAP (Cat No. 311801) was used as the astrocyte marker together with Hs-CHI3L1 (Cat No. 408121). The probes were amplified according to manufacturer’s instructions and labeled with the following red or green color for each experiment. The Hs-CHI3L1 probe was also combined with immunohistochemistry (anti-GFAP and anti-MHCII, Abcam) as described above.

Funding Information:
Lundbeckfonden R118-A11472, OTKA-K77892, Scleroseforeningen R431-A29926-B15690 and R399-A28099-B15690, Region of Southern Denmark 14/24200, Jascha Fonden 5589, Direktør Ejnar Jonasson kaldet Johnsen og hustrus mindelegat 5609, Odense University Hospital (OUH) A474 (to ZI), OTKA-NN109841, GINOP 2.3.2-15-2016-00049. JB is grateful for financial support from JB’s VILLUM Young Investigator Grant nr. 13154. NA received funding from JB’s SDU2020 grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Publisher Copyright:
© 2021, The Author(s).

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Dokumenter og links

10.1038/s41598-021-83591-5Licens: CC BY

Open Access VersionForlagets udgivne version, 1,93 MBLicens: CC BY

Citationsformater

Elkjaer, M. L., Nawrocki, A., Kacprowski, T., Lassen, P., Simonsen, A. H., Marignier, R., Sejbaek, T., Nielsen, H. H., Wermuth, L., Rashid, A. Y., Høgh, P., Sellebjerg, F., Reynolds, R., Baumbach, J., Larsen, M. R., & Illes, Z. (2021). CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes. Scientific Reports, 11, [4132]. https://doi.org/10.1038/s41598-021-83591-5

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title = "CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes",

abstract = "To identify markers in the CSF of multiple sclerosis (MS) subtypes, we used a two-step proteomic approach: (i) Discovery proteomics compared 169 pooled CSF from MS subtypes and inflammatory/degenerative CNS diseases (NMO spectrum and Alzheimer disease) and healthy controls. (ii) Next, 299 proteins selected by comprehensive statistics were quantified in 170 individual CSF samples. (iii) Genes of the identified proteins were also screened among transcripts in 73 MS brain lesions compared to 25 control brains. F-test based feature selection resulted in 8 proteins differentiating the MS subtypes, and secondary progressive (SP)MS was the most different also from controls. Genes of 7 out these 8 proteins were present in MS brain lesions: GOLM was significantly differentially expressed in active, chronic active, inactive and remyelinating lesions, FRZB in active and chronic active lesions, and SELENBP1 in inactive lesions. Volcano maps of normalized proteins in the different disease groups also indicated the highest amount of altered proteins in SPMS. Apolipoprotein C-I, apolipoprotein A-II, augurin, receptor-type tyrosine-protein phosphatase gamma, and trypsin-1 were upregulated in the CSF of MS subtypes compared to controls. This CSF profile and associated brain lesion spectrum highlight non-inflammatory mechanisms in differentiating CNS diseases and MS subtypes and the uniqueness of SPMS.",

author = "Elkjaer, {Maria L.} and Arkadiusz Nawrocki and Tim Kacprowski and Pernille Lassen and Simonsen, {Anja Hviid} and Romain Marignier and Tobias Sejbaek and Nielsen, {Helle H.} and Lene Wermuth and Rashid, {Alyaa Yakut} and Peter H{\o}gh and Finn Sellebjerg and Richard Reynolds and Jan Baumbach and Larsen, {Martin R.} and Zsolt Illes",

note = "Funding Information: Immunohistochemistry and RNAscope of chronic active brain lesion. Human postmortem brain tissue were supplied by the UK Multiple Sclerosis Tissue Bank (UK Multicentre Research Ethics Committee, MREC/02/2/39), funded by the Multiple Sclerosis Society of Great Britain and Northern Ireland (registered charity 207,495). Fresh-frozen blocks containing chronic active lesion from progressive MS patients were sectioned (10-μm), PFA-fixed, blocked in PBS with 10% normal horse serum (NHS) and immunostained with rabbit CHI3L1 (monoclonal antibody) 1:200 (Abcam) followed by biotinylated secondary antibody (Jackson Immu-noresearch Laboratories, Cambridgeshire, UK), avidin/biotin staining (Vector Laboratories, Burlingame, CA) and 3,3′-diaminobenzidine (DAB) staining (Vector Laboratories, Burlingame, CA). The RNAscope 2.5 Duplex Assay (ACD Biosystems) was performed according to the ACD protocol for fresh-frozen tissue. Chronic active lesions were hybridized with two mRNA probes per experiment. Hs-GFAP (Cat No. 311801) was used as the astrocyte marker together with Hs-CHI3L1 (Cat No. 408121). The probes were amplified according to manufacturer{\textquoteright}s instructions and labeled with the following red or green color for each experiment. The Hs-CHI3L1 probe was also combined with immunohistochemistry (anti-GFAP and anti-MHCII, Abcam) as described above. Funding Information: Lundbeckfonden R118-A11472, OTKA-K77892, Scleroseforeningen R431-A29926-B15690 and R399-A28099-B15690, Region of Southern Denmark 14/24200, Jascha Fonden 5589, Direkt{\o}r Ejnar Jonasson kaldet Johnsen og hustrus mindelegat 5609, Odense University Hospital (OUH) A474 (to ZI), OTKA-NN109841, GINOP 2.3.2-15-2016-00049. JB is grateful for financial support from JB{\textquoteright}s VILLUM Young Investigator Grant nr. 13154. NA received funding from JB{\textquoteright}s SDU2020 grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: {\textcopyright} 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

year = "2021",

month = feb,

day = "18",

doi = "10.1038/s41598-021-83591-5",

language = "English",

volume = "11",

journal = "Scientific Reports",

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Elkjaer, ML , Nawrocki, A, Kacprowski, T, Lassen, P, Simonsen, AH, Marignier, R, Sejbaek, T, Nielsen, HH, Wermuth, L, Rashid, AY, Høgh, P, Sellebjerg, F, Reynolds, R, Baumbach, J , Larsen, MR & Illes, Z 2021, 'CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes', Scientific Reports, bind 11, 4132. https://doi.org/10.1038/s41598-021-83591-5

TY - JOUR

T1 - CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes

AU - Elkjaer, Maria L.

AU - Nawrocki, Arkadiusz

AU - Kacprowski, Tim

AU - Lassen, Pernille

AU - Simonsen, Anja Hviid

AU - Marignier, Romain

AU - Sejbaek, Tobias

AU - Nielsen, Helle H.

AU - Wermuth, Lene

AU - Rashid, Alyaa Yakut

AU - Høgh, Peter

AU - Sellebjerg, Finn

AU - Reynolds, Richard

AU - Baumbach, Jan

AU - Larsen, Martin R.

AU - Illes, Zsolt

N1 - Funding Information: Immunohistochemistry and RNAscope of chronic active brain lesion. Human postmortem brain tissue were supplied by the UK Multiple Sclerosis Tissue Bank (UK Multicentre Research Ethics Committee, MREC/02/2/39), funded by the Multiple Sclerosis Society of Great Britain and Northern Ireland (registered charity 207,495). Fresh-frozen blocks containing chronic active lesion from progressive MS patients were sectioned (10-μm), PFA-fixed, blocked in PBS with 10% normal horse serum (NHS) and immunostained with rabbit CHI3L1 (monoclonal antibody) 1:200 (Abcam) followed by biotinylated secondary antibody (Jackson Immu-noresearch Laboratories, Cambridgeshire, UK), avidin/biotin staining (Vector Laboratories, Burlingame, CA) and 3,3′-diaminobenzidine (DAB) staining (Vector Laboratories, Burlingame, CA). The RNAscope 2.5 Duplex Assay (ACD Biosystems) was performed according to the ACD protocol for fresh-frozen tissue. Chronic active lesions were hybridized with two mRNA probes per experiment. Hs-GFAP (Cat No. 311801) was used as the astrocyte marker together with Hs-CHI3L1 (Cat No. 408121). The probes were amplified according to manufacturer’s instructions and labeled with the following red or green color for each experiment. The Hs-CHI3L1 probe was also combined with immunohistochemistry (anti-GFAP and anti-MHCII, Abcam) as described above. Funding Information: Lundbeckfonden R118-A11472, OTKA-K77892, Scleroseforeningen R431-A29926-B15690 and R399-A28099-B15690, Region of Southern Denmark 14/24200, Jascha Fonden 5589, Direktør Ejnar Jonasson kaldet Johnsen og hustrus mindelegat 5609, Odense University Hospital (OUH) A474 (to ZI), OTKA-NN109841, GINOP 2.3.2-15-2016-00049. JB is grateful for financial support from JB’s VILLUM Young Investigator Grant nr. 13154. NA received funding from JB’s SDU2020 grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/2/18

Y1 - 2021/2/18

N2 - To identify markers in the CSF of multiple sclerosis (MS) subtypes, we used a two-step proteomic approach: (i) Discovery proteomics compared 169 pooled CSF from MS subtypes and inflammatory/degenerative CNS diseases (NMO spectrum and Alzheimer disease) and healthy controls. (ii) Next, 299 proteins selected by comprehensive statistics were quantified in 170 individual CSF samples. (iii) Genes of the identified proteins were also screened among transcripts in 73 MS brain lesions compared to 25 control brains. F-test based feature selection resulted in 8 proteins differentiating the MS subtypes, and secondary progressive (SP)MS was the most different also from controls. Genes of 7 out these 8 proteins were present in MS brain lesions: GOLM was significantly differentially expressed in active, chronic active, inactive and remyelinating lesions, FRZB in active and chronic active lesions, and SELENBP1 in inactive lesions. Volcano maps of normalized proteins in the different disease groups also indicated the highest amount of altered proteins in SPMS. Apolipoprotein C-I, apolipoprotein A-II, augurin, receptor-type tyrosine-protein phosphatase gamma, and trypsin-1 were upregulated in the CSF of MS subtypes compared to controls. This CSF profile and associated brain lesion spectrum highlight non-inflammatory mechanisms in differentiating CNS diseases and MS subtypes and the uniqueness of SPMS.

AB - To identify markers in the CSF of multiple sclerosis (MS) subtypes, we used a two-step proteomic approach: (i) Discovery proteomics compared 169 pooled CSF from MS subtypes and inflammatory/degenerative CNS diseases (NMO spectrum and Alzheimer disease) and healthy controls. (ii) Next, 299 proteins selected by comprehensive statistics were quantified in 170 individual CSF samples. (iii) Genes of the identified proteins were also screened among transcripts in 73 MS brain lesions compared to 25 control brains. F-test based feature selection resulted in 8 proteins differentiating the MS subtypes, and secondary progressive (SP)MS was the most different also from controls. Genes of 7 out these 8 proteins were present in MS brain lesions: GOLM was significantly differentially expressed in active, chronic active, inactive and remyelinating lesions, FRZB in active and chronic active lesions, and SELENBP1 in inactive lesions. Volcano maps of normalized proteins in the different disease groups also indicated the highest amount of altered proteins in SPMS. Apolipoprotein C-I, apolipoprotein A-II, augurin, receptor-type tyrosine-protein phosphatase gamma, and trypsin-1 were upregulated in the CSF of MS subtypes compared to controls. This CSF profile and associated brain lesion spectrum highlight non-inflammatory mechanisms in differentiating CNS diseases and MS subtypes and the uniqueness of SPMS.

U2 - 10.1038/s41598-021-83591-5

DO - 10.1038/s41598-021-83591-5

M3 - Journal article

C2 - 33603109

AN - SCOPUS:85101239393

SN - 2045-2322

VL - 11

JO - Scientific Reports

JF - Scientific Reports

M1 - 4132

ER -

CSF proteome in multiple sclerosis subtypes related to brain lesion transcriptomes

Abstrakt

Bibliografisk note

Dokumenter og links

Fingeraftryk

Citationsformater