Experimental demyelination and axonal loss are reduced in MicroRNA-146a deficient mice

Nellie A. Martin, Viktor Molnar, Gabor T. Szilagyi, Maria L. Elkjaer, Arkadiusz Nawrocki, Justyna Okarmus, Agnieszka Wlodarczyk, Eva K. Thygesen, Miklos Palkovits, Ferenc Gallyas, Martin R. Larsen, Hans Lassmann, Eirikur Benedikz, Trevor Owens, Asa F. Svenningsen, Zsolt Illes*

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Background: The cuprizone (CPZ) model of multiple sclerosis (MS) was used to identify microRNAs (miRNAs) related to in vivo de- and remyelination. We further investigated the role of miR-146a in miR-146a-deficient (KO) mice: this miRNA is differentially expressed in MS lesions and promotes differentiation of oligodendrocyte precursor cells (OPCs) during remyelination, but its role has not been examined during demyelination. Methods: MicroRNAs were examined by Agilent Mouse miRNA Microarray in the corpus callosum during CPZ-induced demyelination and remyelination. Demyelination, axonal loss, changes in number of oligodendrocytes, OPCs, and macrophages/microglia was compared by histology/immunohistochemistry between KO and WT mice. Differential expression of target genes and proteins of miR-146a was analyzed in the transcriptome (4 × 44K Agilent Whole Mouse Genome Microarray) and proteome (liquid chromatography tandem mass spectrometry) of CPZ-induced de- and remyelination in WT mice. Levels of proinflammatory molecules in the corpus callosum were compared in WT versus KO mice by Meso Scale Discovery multiplex protein analysis. Results: miR-146a was increasingly upregulated during CPZ-induced de- and remyelination. The absence of miR-146a in KO mice protected against demyelination, axonal loss, body weight loss, and atrophy of thymus and spleen. The number of CNP+ oligodendrocytes was increased during demyelination in the miR-146a KO mice, while there was a trend of increased number of NG2+ OPCs in the WT mice. miR-146a target genes, SNAP25 and SMAD4, were downregulated in the proteome of demyelinating corpus callosum in WT mice. Higher levels of SNAP25 were measured by ELISA in the corpus callosum of miR-146a KO mice, but there was no difference between KO and WT mice during demyelination. Multiplex protein analysis of the corpus callosum lysate revealed upregulated TNF-RI, TNF-RII, and CCL2 in the WT mice in contrast to KO mice. The number of Mac3+ and Iba1+ macrophages/microglia was reduced in the demyelinating corpus callosum of the KO mice. Conclusion: During demyelination, absence of miR-146a reduced inflammatory responses, demyelination, axonal loss, the number of infiltrating macrophages, and increased the number of myelinating oligodendrocytes. The number of OPCs was slightly higher in the WT mice during remyelination, indicating a complex role of miR-146a during in vivo de- and remyelination.

OriginalsprogEngelsk
Artikelnummer490
TidsskriftFrontiers in Immunology
Vol/bind9
Antal sider14
ISSN1664-3224
DOI
StatusUdgivet - 2018

Fingeraftryk

Demyelinating Diseases
Oligodendroglia
Corpus Callosum
Cuprizone
MicroRNAs
Macrophages
Microglia
Proteome
Mouse Mirn146 microRNA
Receptors, Tumor Necrosis Factor, Type II
Proteins
Tandem Mass Spectrometry
Transcriptome
Liquid Chromatography
Down-Regulation

Citer dette

@article{ca3ba90f2274474e830a57b104fd7aba,
title = "Experimental demyelination and axonal loss are reduced in MicroRNA-146a deficient mice",
abstract = "Background: The cuprizone (CPZ) model of multiple sclerosis (MS) was used to identify microRNAs (miRNAs) related to in vivo de- and remyelination. We further investigated the role of miR-146a in miR-146a-deficient (KO) mice: this miRNA is differentially expressed in MS lesions and promotes differentiation of oligodendrocyte precursor cells (OPCs) during remyelination, but its role has not been examined during demyelination. Methods: MicroRNAs were examined by Agilent Mouse miRNA Microarray in the corpus callosum during CPZ-induced demyelination and remyelination. Demyelination, axonal loss, changes in number of oligodendrocytes, OPCs, and macrophages/microglia was compared by histology/immunohistochemistry between KO and WT mice. Differential expression of target genes and proteins of miR-146a was analyzed in the transcriptome (4 × 44K Agilent Whole Mouse Genome Microarray) and proteome (liquid chromatography tandem mass spectrometry) of CPZ-induced de- and remyelination in WT mice. Levels of proinflammatory molecules in the corpus callosum were compared in WT versus KO mice by Meso Scale Discovery multiplex protein analysis. Results: miR-146a was increasingly upregulated during CPZ-induced de- and remyelination. The absence of miR-146a in KO mice protected against demyelination, axonal loss, body weight loss, and atrophy of thymus and spleen. The number of CNP+ oligodendrocytes was increased during demyelination in the miR-146a KO mice, while there was a trend of increased number of NG2+ OPCs in the WT mice. miR-146a target genes, SNAP25 and SMAD4, were downregulated in the proteome of demyelinating corpus callosum in WT mice. Higher levels of SNAP25 were measured by ELISA in the corpus callosum of miR-146a KO mice, but there was no difference between KO and WT mice during demyelination. Multiplex protein analysis of the corpus callosum lysate revealed upregulated TNF-RI, TNF-RII, and CCL2 in the WT mice in contrast to KO mice. The number of Mac3+ and Iba1+ macrophages/microglia was reduced in the demyelinating corpus callosum of the KO mice. Conclusion: During demyelination, absence of miR-146a reduced inflammatory responses, demyelination, axonal loss, the number of infiltrating macrophages, and increased the number of myelinating oligodendrocytes. The number of OPCs was slightly higher in the WT mice during remyelination, indicating a complex role of miR-146a during in vivo de- and remyelination.",
keywords = "Cuprizone, Demyelination, MiR-146a, MiR-181b, MiR-193a, Multiple sclerosis, Remyelination, Demyelinating Diseases/genetics, MicroRNAs/genetics, Humans, Mice, Inbred C57BL, Axons/pathology, Corpus Callosum/physiology, Gene Expression Profiling, Oligodendroglia/physiology, Mice, Knockout, Animals, Chemokine CCL2/genetics, Female, Models, Animal, Cell Differentiation, Mice, Receptors, Tumor Necrosis Factor/genetics",
author = "Martin, {Nellie A.} and Viktor Molnar and Szilagyi, {Gabor T.} and Elkjaer, {Maria L.} and Arkadiusz Nawrocki and Justyna Okarmus and Agnieszka Wlodarczyk and Thygesen, {Eva K.} and Miklos Palkovits and Ferenc Gallyas and Larsen, {Martin R.} and Hans Lassmann and Eirikur Benedikz and Trevor Owens and Svenningsen, {Asa F.} and Zsolt Illes",
year = "2018",
doi = "10.3389/fimmu.2018.00490",
language = "English",
volume = "9",
journal = "Frontiers in Immunology",
issn = "1664-3224",
publisher = "Frontiers Research Foundation",

}

Experimental demyelination and axonal loss are reduced in MicroRNA-146a deficient mice. / Martin, Nellie A.; Molnar, Viktor; Szilagyi, Gabor T.; Elkjaer, Maria L.; Nawrocki, Arkadiusz; Okarmus, Justyna; Wlodarczyk, Agnieszka; Thygesen, Eva K.; Palkovits, Miklos; Gallyas, Ferenc; Larsen, Martin R.; Lassmann, Hans; Benedikz, Eirikur; Owens, Trevor; Svenningsen, Asa F.; Illes, Zsolt.

I: Frontiers in Immunology, Bind 9, 490, 2018.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Experimental demyelination and axonal loss are reduced in MicroRNA-146a deficient mice

AU - Martin, Nellie A.

AU - Molnar, Viktor

AU - Szilagyi, Gabor T.

AU - Elkjaer, Maria L.

AU - Nawrocki, Arkadiusz

AU - Okarmus, Justyna

AU - Wlodarczyk, Agnieszka

AU - Thygesen, Eva K.

AU - Palkovits, Miklos

AU - Gallyas, Ferenc

AU - Larsen, Martin R.

AU - Lassmann, Hans

AU - Benedikz, Eirikur

AU - Owens, Trevor

AU - Svenningsen, Asa F.

AU - Illes, Zsolt

PY - 2018

Y1 - 2018

N2 - Background: The cuprizone (CPZ) model of multiple sclerosis (MS) was used to identify microRNAs (miRNAs) related to in vivo de- and remyelination. We further investigated the role of miR-146a in miR-146a-deficient (KO) mice: this miRNA is differentially expressed in MS lesions and promotes differentiation of oligodendrocyte precursor cells (OPCs) during remyelination, but its role has not been examined during demyelination. Methods: MicroRNAs were examined by Agilent Mouse miRNA Microarray in the corpus callosum during CPZ-induced demyelination and remyelination. Demyelination, axonal loss, changes in number of oligodendrocytes, OPCs, and macrophages/microglia was compared by histology/immunohistochemistry between KO and WT mice. Differential expression of target genes and proteins of miR-146a was analyzed in the transcriptome (4 × 44K Agilent Whole Mouse Genome Microarray) and proteome (liquid chromatography tandem mass spectrometry) of CPZ-induced de- and remyelination in WT mice. Levels of proinflammatory molecules in the corpus callosum were compared in WT versus KO mice by Meso Scale Discovery multiplex protein analysis. Results: miR-146a was increasingly upregulated during CPZ-induced de- and remyelination. The absence of miR-146a in KO mice protected against demyelination, axonal loss, body weight loss, and atrophy of thymus and spleen. The number of CNP+ oligodendrocytes was increased during demyelination in the miR-146a KO mice, while there was a trend of increased number of NG2+ OPCs in the WT mice. miR-146a target genes, SNAP25 and SMAD4, were downregulated in the proteome of demyelinating corpus callosum in WT mice. Higher levels of SNAP25 were measured by ELISA in the corpus callosum of miR-146a KO mice, but there was no difference between KO and WT mice during demyelination. Multiplex protein analysis of the corpus callosum lysate revealed upregulated TNF-RI, TNF-RII, and CCL2 in the WT mice in contrast to KO mice. The number of Mac3+ and Iba1+ macrophages/microglia was reduced in the demyelinating corpus callosum of the KO mice. Conclusion: During demyelination, absence of miR-146a reduced inflammatory responses, demyelination, axonal loss, the number of infiltrating macrophages, and increased the number of myelinating oligodendrocytes. The number of OPCs was slightly higher in the WT mice during remyelination, indicating a complex role of miR-146a during in vivo de- and remyelination.

AB - Background: The cuprizone (CPZ) model of multiple sclerosis (MS) was used to identify microRNAs (miRNAs) related to in vivo de- and remyelination. We further investigated the role of miR-146a in miR-146a-deficient (KO) mice: this miRNA is differentially expressed in MS lesions and promotes differentiation of oligodendrocyte precursor cells (OPCs) during remyelination, but its role has not been examined during demyelination. Methods: MicroRNAs were examined by Agilent Mouse miRNA Microarray in the corpus callosum during CPZ-induced demyelination and remyelination. Demyelination, axonal loss, changes in number of oligodendrocytes, OPCs, and macrophages/microglia was compared by histology/immunohistochemistry between KO and WT mice. Differential expression of target genes and proteins of miR-146a was analyzed in the transcriptome (4 × 44K Agilent Whole Mouse Genome Microarray) and proteome (liquid chromatography tandem mass spectrometry) of CPZ-induced de- and remyelination in WT mice. Levels of proinflammatory molecules in the corpus callosum were compared in WT versus KO mice by Meso Scale Discovery multiplex protein analysis. Results: miR-146a was increasingly upregulated during CPZ-induced de- and remyelination. The absence of miR-146a in KO mice protected against demyelination, axonal loss, body weight loss, and atrophy of thymus and spleen. The number of CNP+ oligodendrocytes was increased during demyelination in the miR-146a KO mice, while there was a trend of increased number of NG2+ OPCs in the WT mice. miR-146a target genes, SNAP25 and SMAD4, were downregulated in the proteome of demyelinating corpus callosum in WT mice. Higher levels of SNAP25 were measured by ELISA in the corpus callosum of miR-146a KO mice, but there was no difference between KO and WT mice during demyelination. Multiplex protein analysis of the corpus callosum lysate revealed upregulated TNF-RI, TNF-RII, and CCL2 in the WT mice in contrast to KO mice. The number of Mac3+ and Iba1+ macrophages/microglia was reduced in the demyelinating corpus callosum of the KO mice. Conclusion: During demyelination, absence of miR-146a reduced inflammatory responses, demyelination, axonal loss, the number of infiltrating macrophages, and increased the number of myelinating oligodendrocytes. The number of OPCs was slightly higher in the WT mice during remyelination, indicating a complex role of miR-146a during in vivo de- and remyelination.

KW - Cuprizone

KW - Demyelination

KW - MiR-146a

KW - MiR-181b

KW - MiR-193a

KW - Multiple sclerosis

KW - Remyelination

KW - Demyelinating Diseases/genetics

KW - MicroRNAs/genetics

KW - Humans

KW - Mice, Inbred C57BL

KW - Axons/pathology

KW - Corpus Callosum/physiology

KW - Gene Expression Profiling

KW - Oligodendroglia/physiology

KW - Mice, Knockout

KW - Animals

KW - Chemokine CCL2/genetics

KW - Female

KW - Models, Animal

KW - Cell Differentiation

KW - Mice

KW - Receptors, Tumor Necrosis Factor/genetics

U2 - 10.3389/fimmu.2018.00490

DO - 10.3389/fimmu.2018.00490

M3 - Journal article

C2 - 29593734

AN - SCOPUS:85043452033

VL - 9

JO - Frontiers in Immunology

JF - Frontiers in Immunology

SN - 1664-3224

M1 - 490

ER -