title: Integrative proteomics, genomics, and translational immunology approaches reveal mutated forms of Proteolipid Protein 1 (PLP1) and mutant-specific immune response in multiple sclerosis.
abstract: In order to gain mechanistic insights into multiple sclerosis (MS) pathogenesis, we utilized a multi-dimensional approach to test the hypothesis that mutations in myelin proteins lead to immune activation and central nervous system autoimmunity in MS. Mass spectrometry-based proteomic analysis of human MS brain lesions revealed seven unique mutations of PLP1; a key myelin protein that is known to be destroyed in MS. Surprisingly, in-depth genomic analysis of two MS patients at the genomic DNA and mRNA confirmed mutated PLP1 in RNA, but not in the genomic DNA. Quantification of wild type and mutant PLP RNA levels by qPCR further validated the presence of mutant PLP RNA in the MS patients. To seek evidence linking mutations in abundant myelin proteins and immune-mediated destruction of myelin, specific immune response against mutant PLP1 in MS patients was examined. Thus, we have designed paired, wild type and mutant peptide microarrays, and examined antibody response to multiple mutated PLP1 in sera from MS patients. Consistent with the idea of different patients exhibiting unique mutation profiles, we found that 13 out of 20 MS patients showed antibody responses against specific but not against all the mutant-PLP1 peptides. Interestingly, we found mutant PLP-directed antibody response against specific mutant peptides in the sera of pre-MS controls. The results from integrative proteomic, genomic, and immune analyses reveal a possible mechanism of mutation-driven pathogenesis in human MS. The study also highlights the need for integrative genomic and proteomic analyses for uncovering pathogenic mechanisms of human diseases.
authors.affiliation: Department of Cell Biology, Center for Vascular Biology, University of Connecticut School of Medicine, Farmington, CT, USA.
authors.forename: Veneta
authors.initials: V
authors.lastname: Qendro
authors.affiliation: Department of Cell Biology, Center for Vascular Biology, University of Connecticut School of Medicine, Farmington, CT, USA.
authors.forename: Grace A
authors.initials: GA
authors.lastname: Bugos
authors.affiliation: Department of Cell Biology, Center for Vascular Biology, University of Connecticut School of Medicine, Farmington, CT, USA.
authors.forename: Debbie H
authors.initials: DH
authors.lastname: Lundgren
authors.affiliation: Molecular Core Facility, University of Connecticut School of Medicine, Farmington, CT, USA.
authors.forename: John
authors.initials: J
authors.lastname: Glynn
authors.affiliation: Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: May H
authors.initials: MH
authors.lastname: Han
authors.affiliation: Department of Cell Biology, Center for Vascular Biology, University of Connecticut School of Medicine, Farmington, CT, USA.
authors.forename: David K
authors.initials: DK
authors.lastname: Han
mesh_terms: D000486:Allergy and Immunology; D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology; D005260:Female; D006801:Humans; D008954:Models, Biological; D009103:Multiple Sclerosis / Q000276:immunology* / Q000378:metabolism*; D009154:Mutation / Q000235:genetics*; D018991:Myelin Proteolipid Protein / Q000737:chemistry / Q000235:genetics*; D040901:Proteomics / Q000379:methods*; D057170:Translational Research, Biomedical / Q000379:methods*
chemical_list: D000906:Antibodies; D018991:Myelin Proteolipid Protein; C472930:PLP1 protein, human
keywords: Genomics; Immunoproteomics; Multiple sclerosis; Next generation sequencing; PLP1; Peptide microarrays; Precision; Proteomics