Contributions
Abstract: P833
Type: Poster
Abstract Category: Clinical aspects of MS - 1 Diagnosis and differential diagnosis
CLIPPERS is a rare syndrome with relapsing brainstem/cerebellar symptoms.
To identify pathogenic mechanisms underlying CLIPPERS and to establish potential biomarkers, we compared the proteome of 5 CSF samples from 4 CLIPPERS patients to CSF from 5 patients with Alzheimer disease (AD) by liquid chromatography mass spectrometry. 207 proteins in the CSF could discriminate CLIPPERS from AD. The complement cascade, immunoglobulins, and matrix proteins were among the most frequently represented pathways. Network and ingenuity pathway analysis of upstream regulators suggested the importance of VCAM1, IFN-gamma, IL-1 and IL-10 in dysregulated CSF pathways in CLIPPERS. Immunohistochemistry on 3 brain samples confirmed the perivascular complement activation i.e. deposition of C3bc, C3d and the terminal C5b-9 complex that partially overlapped with accumulation of IgG in the vessel wall. Besides endothelial cell damage, reactivity to smooth muscle actin was lost in the walls of inflamed vessels, but the glia limitans was preserved. Activation of complement system was further suggested by elevated concentrations of C3 activation product C3c (C3bc) and the soluble terminal C5b-9 complement complex in 2 CSF samples of one patient. To investigate specific IgG binding to CNS structures, we incubated a pool of 4 CSF samples or isolated serum IgG from 3 CLIPPERS patients with rat brain tissue or formalin-fixed brain tissue of another CLIPPERS patient, but we could not identify specific IgG staining. To further support proteome data as possible biomarkers, we compared inflammatory and vessel-associated proteins in the CSF of 5 patients with CLIPPERS, 7 healthy subjects (HS), and 9 patients with RRMS. Semiquantitative array of 40 biomarkers indicated that elevated concentrations of interleukin-8 (IL-8, CXCL8), eotaxin (CCL11) and the granulocyte-colony stimulating factor (GCSF) in the CSF could distinguish CLIPPERS from HS. By investigating 37 biomarkers in the CSF by quantitative MesoScale array, we found that increased concentration of IL-8 (CXCL8), ICAM1 and VCAM1 were able to differentiate CLIPPERS from both HS and RRMS.
In conclusion, CSF proteome and brain autopsy/biopsy data suggest that IgG deposition, alterations of the extracellular matrix, and complement activation contribute to the perivascular inflammation in CLIPPERS. Importantly, VCAM1, ICAM1 and IL-8 in the CSF may differentiate CLIPPERS from RRMS.
Disclosure: The study was funded by Lundbeckfonden R118-A11472, Jascha Fonden 5589, Scleroseforeningen R399-A28099-B15690 and R431-A29926, Region of Southern Denmark 14/24200, Direktør Ejnar Jonasson kaldet Johnsen og hustrus mindelegat 5609, Odense University Hospital R9-A559 and A474 (Denmark) to ZI. The Danish Research Council of Independent Research (DFF-6110-00489) to PG. Deutsche Forschungsgemeinschaft (Exc 257) and the German Ministry for Education and Research (BMBF Competence Network Multiple Sclerosis) to FP. German Ministry for Education and Research (BMBF Competence Network Multiple Sclerosis) to KR. This work was supported by a generous grant from the VILLUM Foundation to the VILLUM Center for Bioanalytical Sciences at the University of Southern Denmark. This study is not sponsored by industry.
Dr. Illes reports grants from Lundbeckfonden, grants from Scleroseforeningen, during the conduct of the study; personal fees from Biogen Idec, grants and personal fees from Sanofi Genzyme, personal fees from Novartis, personal fees from Merck Serono, outside the submitted work;
Dr. Lassmann reports personal fees from Novartis, personal fees from Sanofi Aventis, personal fees from TEVA, personal fees from Roche, outside the submitted work;
Dr. Paul reports grants and personal fees from Various pharmaceutical companies, outside the submitted work;
Dr. Ruprecht reports grants from German Ministry of Education and Research (BMBF/KKNMS, Competence Network Multiple Sclerosis), during the conduct of the study; grants and personal fees from Novartis, personal fees from Bayer Healthcare, personal fees from Biogen Idec, grants and personal fees from Merck Serono, personal fees from sanofi-aventis/Genzyme, personal fees from Teva Pharmaceuticals, grants from Guthy Jackson Charitable Foundation, outside the submitted work.
Abstract: P833
Type: Poster
Abstract Category: Clinical aspects of MS - 1 Diagnosis and differential diagnosis
CLIPPERS is a rare syndrome with relapsing brainstem/cerebellar symptoms.
To identify pathogenic mechanisms underlying CLIPPERS and to establish potential biomarkers, we compared the proteome of 5 CSF samples from 4 CLIPPERS patients to CSF from 5 patients with Alzheimer disease (AD) by liquid chromatography mass spectrometry. 207 proteins in the CSF could discriminate CLIPPERS from AD. The complement cascade, immunoglobulins, and matrix proteins were among the most frequently represented pathways. Network and ingenuity pathway analysis of upstream regulators suggested the importance of VCAM1, IFN-gamma, IL-1 and IL-10 in dysregulated CSF pathways in CLIPPERS. Immunohistochemistry on 3 brain samples confirmed the perivascular complement activation i.e. deposition of C3bc, C3d and the terminal C5b-9 complex that partially overlapped with accumulation of IgG in the vessel wall. Besides endothelial cell damage, reactivity to smooth muscle actin was lost in the walls of inflamed vessels, but the glia limitans was preserved. Activation of complement system was further suggested by elevated concentrations of C3 activation product C3c (C3bc) and the soluble terminal C5b-9 complement complex in 2 CSF samples of one patient. To investigate specific IgG binding to CNS structures, we incubated a pool of 4 CSF samples or isolated serum IgG from 3 CLIPPERS patients with rat brain tissue or formalin-fixed brain tissue of another CLIPPERS patient, but we could not identify specific IgG staining. To further support proteome data as possible biomarkers, we compared inflammatory and vessel-associated proteins in the CSF of 5 patients with CLIPPERS, 7 healthy subjects (HS), and 9 patients with RRMS. Semiquantitative array of 40 biomarkers indicated that elevated concentrations of interleukin-8 (IL-8, CXCL8), eotaxin (CCL11) and the granulocyte-colony stimulating factor (GCSF) in the CSF could distinguish CLIPPERS from HS. By investigating 37 biomarkers in the CSF by quantitative MesoScale array, we found that increased concentration of IL-8 (CXCL8), ICAM1 and VCAM1 were able to differentiate CLIPPERS from both HS and RRMS.
In conclusion, CSF proteome and brain autopsy/biopsy data suggest that IgG deposition, alterations of the extracellular matrix, and complement activation contribute to the perivascular inflammation in CLIPPERS. Importantly, VCAM1, ICAM1 and IL-8 in the CSF may differentiate CLIPPERS from RRMS.
Disclosure: The study was funded by Lundbeckfonden R118-A11472, Jascha Fonden 5589, Scleroseforeningen R399-A28099-B15690 and R431-A29926, Region of Southern Denmark 14/24200, Direktør Ejnar Jonasson kaldet Johnsen og hustrus mindelegat 5609, Odense University Hospital R9-A559 and A474 (Denmark) to ZI. The Danish Research Council of Independent Research (DFF-6110-00489) to PG. Deutsche Forschungsgemeinschaft (Exc 257) and the German Ministry for Education and Research (BMBF Competence Network Multiple Sclerosis) to FP. German Ministry for Education and Research (BMBF Competence Network Multiple Sclerosis) to KR. This work was supported by a generous grant from the VILLUM Foundation to the VILLUM Center for Bioanalytical Sciences at the University of Southern Denmark. This study is not sponsored by industry.
Dr. Illes reports grants from Lundbeckfonden, grants from Scleroseforeningen, during the conduct of the study; personal fees from Biogen Idec, grants and personal fees from Sanofi Genzyme, personal fees from Novartis, personal fees from Merck Serono, outside the submitted work;
Dr. Lassmann reports personal fees from Novartis, personal fees from Sanofi Aventis, personal fees from TEVA, personal fees from Roche, outside the submitted work;
Dr. Paul reports grants and personal fees from Various pharmaceutical companies, outside the submitted work;
Dr. Ruprecht reports grants from German Ministry of Education and Research (BMBF/KKNMS, Competence Network Multiple Sclerosis), during the conduct of the study; grants and personal fees from Novartis, personal fees from Bayer Healthcare, personal fees from Biogen Idec, grants and personal fees from Merck Serono, personal fees from sanofi-aventis/Genzyme, personal fees from Teva Pharmaceuticals, grants from Guthy Jackson Charitable Foundation, outside the submitted work.