WO2017070589A1 - Méthodes et compositions pour le diagnostic et le traitement de maladies autoimmunes du système nerveux central - Google Patents

Méthodes et compositions pour le diagnostic et le traitement de maladies autoimmunes du système nerveux central Download PDF

Info

Publication number
WO2017070589A1
WO2017070589A1 PCT/US2016/058285 US2016058285W WO2017070589A1 WO 2017070589 A1 WO2017070589 A1 WO 2017070589A1 US 2016058285 W US2016058285 W US 2016058285W WO 2017070589 A1 WO2017070589 A1 WO 2017070589A1
Authority
WO
WIPO (PCT)
Prior art keywords
type
expression
eae
cxcr2
subject
Prior art date
Application number
PCT/US2016/058285
Other languages
English (en)
Inventor
Mari L. SHINOHARA
Makoto Inoue
Original Assignee
Duke University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Duke University filed Critical Duke University
Publication of WO2017070589A1 publication Critical patent/WO2017070589A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/521—Chemokines
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/715—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons
    • G01N2333/7151—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons for tumor necrosis factor [TNF]; for lymphotoxin [LT]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/715—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons
    • G01N2333/7158—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons for chemokines
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/285—Demyelinating diseases; Multipel sclerosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • This disclosure relates to central nervous system autoimmune diseases such as multiple sclerosis and diagnosis and treatment thereof.
  • MS Multiple sclerosis
  • CNS central nervous system
  • ⁇ a first-line treatment to MS patients for over 15 years in progressive and relapsing-remitting MS (RRMS)
  • RRMS progressive and relapsing-remitting MS
  • the disclosure relates to methods of treating a central nervous system (CNS) autoimmune disease in a subject.
  • the method includes administering to the subject at least one of an LT ⁇ R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • the inhibitor includes siRNA, an antibody, or a small molecule.
  • the CNS autoimmune disease includes multiple sclerosis (MS).
  • the MS is not responsive to treatment with ⁇ .
  • the MS includes NLRP3-independent MS.
  • the MS includes Type B MS.
  • the subject has irreversible neuronal damage, demyelination, or a combination thereof.
  • the irreversible neuronal damage, demyelination, or a combination thereof is in the brain or optic nerve.
  • the Type B MS is characterized by at least one of: irreversible neuronal damage, demyelination, or a combination thereof; irresponsiveness to treatment with ⁇ ; lack of activation of the NLRP3 inflammasome; increased expression of Ltbr, CXCR2, CXCR1 , or Sema6B; increased levels of B cells in the DLNs; or a combination thereof, relative to a control.
  • the expression of CXCR2 is increased in neutrophils, CD4+ cells, or a combination thereof.
  • the disclosure relates to methods of diagnosing Type B MS in a subject.
  • the method includes: determining the expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in a sample from the subject, wherein an increase of the expression relative to a control indicates a diagnosis that the subject is afflicted with Type B MS.
  • the method further includes diagnosing the patient as having Type B MS when the expression is increased relative to the control.
  • the method further includes administering treatment with one or more pharmaceutical compositions, to the subject diagnosed as having Type B MS.
  • the treatment includes at least one of an LT ⁇ R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • Another aspect of the disclosure provides methods of diagnosing Type B MS in a subject.
  • the method includes: (a) contacting a sample from the subject with an agent for specifically detecting expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (b) determining a level of expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; and (c) comparing the level of expression in the sample with a control level of expression to diagnose Type B MS.
  • the agent includes a polynucleotide probe that hybridizes specifically with RNA transcribed from gene encoding at least one of CXCR2, CXCR1 , Ltbr, and Sema6B present in the subject.
  • the agent includes an antibody that binds to at least one of CXCR2, CXCR1 , Ltbr, and Sema6B.
  • the methods further include diagnosing the patient as having Type B MS when the level of expression is increased relative to the control.
  • the methods further include administering treatment with one or more
  • the treatment includes at least one of an LT ⁇ R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • the disclosure relates to methods of predicting responsiveness of a subject with MS to treatment with ⁇ .
  • the method includes (a) contacting a sample from the subject with an agent for specifically detecting expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (b) determining a level of expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (c) comparing the level of expression in the sample with a control level of expression; and (d) determining that the subject is not responsive to treatment with ⁇ when the level of expression is increased, and that the subject is responsive to treatment with ⁇ when the level of expression is the same or decreased.
  • the method further includes administering treatment with one or more pharmaceutical compositions, to the subject determined to be not responsive to treatment with ⁇ .
  • the treatment may include at least one of an LT ⁇ R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • the method further includes administering treatment with one or more pharmaceutical compositions, to the subject determined to be responsive to treatment with ⁇ .
  • the treatment may include ⁇ .
  • the expression is measured by determining at least one of mRNA levels, protein levels, or protein activity levels. In some embodiments, the determining step includes at least one of PCR or antibody binding. In some embodiments, the sample includes peripheral blood mononuclear cells (PBMCs).
  • PBMCs peripheral blood mononuclear cells
  • FIG. 1 shows that type-B EAE develops independent of the NLRP3 inflammasome and is resistant to ⁇ treatment.
  • D, E Serum levels of I L-1 ⁇ and ⁇ in naive mice, Type-A and Type-B EAE mice at 9-dpi.
  • G EAE scores of WT and Nlrp3-1 mice immunized with a Type-A EAE induction method with MHV68 infection (left panel), and WT Type-A EAE mice infected with MHV68 with or without ⁇ treatment (right panel).
  • FIG. 2 shows that ml_T is a key initiator of Type-B EAE.
  • A Expression levels of Lta mRNA) in DCs obtained front DLNs of naive mice or mice at 9 dpi.
  • B Percentages of DCs, positive for cell surface LTa expression, in DLNs of naive mice or mice with EAE at 9-dpi, determined by flow cytometry.
  • C Methylation analysis by bisulfite conversion with DCs obtained from DLNs, pooled from eight WT mice, Type-A and Type-B EAE at 9-dpi. Methylated and unmethylated CpG were shown with black and gray boxes, respectively.
  • (D) EAE scores of WT mice after Type-A and Type-B induction with or without LT ⁇ R-Fc treatment (n 5-9).
  • FIG. 3 Gene expression profile of CD4+ T cells between Type-A and Type-B.
  • CD4+ T cells were FACS-purified from spleens at 9-dpi and submitted for the RNA-seq analysis.
  • A Heatmap of 1846 differentially expressed genes (P ⁇ 0.05; log2FC >l or ⁇ -1) between CD4+ T cells from Type-A and Type-B EAE mice.
  • B Scatter plot comparing transcriptome.
  • C C
  • FIG. 4 Expression of Cxcr2, Cxcrl , and Ltbr to characterize ⁇ - ⁇ 3 ⁇ 3 ⁇ 3 ⁇ EAE and MS.
  • B mRNA levels of Cxcr2, Cxcrl , and Ltbr in splenic CD4+ T cells obtained from naive mice and mice with Type-A or Type-B EAE induction at 9-dpi determined by qPCR.
  • C Levels of Cxcr2, Cxcrl , and Ltbr mRNA in CD4+ T 6 h after rLT1 treatment at indicated concentrations.
  • D Proportions of CXCR2-positive cells and MFI of CXCR2 staining in splenic CD+ T cells obtained from naive mice and mice with Type-A or Type- B EAE induction at 9-dpi determined by flow, cytometry.
  • E Relative expression of Cxcr2, Cxcrl , and Ltbr normalized by Vcaml in total PBMCs from IFIS ⁇ -responder and non-responder RRMS patients. One circle denotes one patient (*; p ⁇ 0.05).
  • FIG. 5 Involvement of the CXCL1 and CXCR2 in Type-B EAE development.
  • B Levels of CXCL1 in serum from naive mice or mice with Type-A or Type-B EAE mice on 9-dpi determined by ELISA.
  • C CXCL1 staining in the choroid plexus on 9-dpi.
  • D Levels of CXCL1 in serum from Type-B EAE with or without LT ⁇ R-Fc treatment on 9 dpi.
  • F EAE score in Lck-Cre Cxcr2 l mice (CXCR2-cKO) and Cxcr2 fl fl mice (control)(*; p ⁇ 0.05).
  • FIG. 6 Persistent disease severity and spinal transected neurites in Type-B EAE mice.
  • B B
  • FIG. 7. Type-B EAE-derived T cells cause neuronal retraction.
  • A-C CD4+ T cells were obtained from spleens of either naive, Type-A EAE, or Type-B EAE mice; then, co-cultured with neurons. Representative images of hippocampal neuron stained with MAP2 (red) and DAPI (blue) one day after co-culture with indicated CD4+ T cells (A). Scale bars denote 50 ⁇ .
  • B Average numbers of axon and dendrite branches per neuron (left panel) and distributions of branch numbers (right panel)(C).
  • E, F CD4+ T cells were obtained from either naive or Type-B EAE mice, and Sema6b mRNA was knocked down by shRNA in the indicated group. T cells were then co-cultured with neurons. Representative images of neurons stained with MAP2 (red) and DAPI (blue)(E). Average neurite lengths (F, left panel) and average numbers of axon and dendrite branches per neuron (F, right panel).
  • Results are expressed as the mean ⁇ SEM by three independent experiments.
  • G Expression levels of Sema6b mRNA in CD4+ T cells isolated from Type-A EAE mice with or without rLT treatment for 6 h.
  • FIG. 8 EAE induced by several immunization methods and phenotypic effects on cell migration, demyelination and nociceptive sensitivity in mice with type A and type B EAE.
  • (C) Numbers of total immune cells were evaluated in the brains and spinal cords (S.C.) of mice with Type-B EAE at 17-dpi. n 4.
  • F T2 FLAIR MRI analysis of spinal cords obtained from mice at 18-dpi. Yellow arrows indicate areas of potential myelin loss.
  • A Flow charts showing ml_T expression in DC (CD1 1c + gated) obtained from naive mice, Type-A EAE mice, and Type-B EAE mice at 9-dpi.
  • FIG. 11 Comparison of cell populations and gene expression in mouse and RRMS patients who have diseases that do not respond to ⁇ therapies.
  • FIG. 12 CXCL1 expression in spinal cords. Shown are representative images of typical CXCL1 staining in spinal cords from naive mice and mice with either Type-A or Type-B EAE at 9-dpi (scale bars, 200 ⁇ ). Images are representatives from 3 similar experiments.
  • FIG. 13 Histology of spinal cords in mice with type A or type B EAE. Shown are representative images of typical staining from multiple mice.
  • A H/E staining in the spinal cord of Type-A and Type-B EAE mice at 70-dpi.
  • B IGF-1 staining in spinal cord of naive mice and mice with either Type-A or Type-B EAE at 22-dpi.
  • C D
  • FIG. 15 Schematic diagram of the distinct pathology of the two EAE subtypes.
  • Type-A EAE is NLRP3 inflammasome-dependent and ⁇ - ⁇ .
  • Type-B EAE is induced by immunization with higher doses of Mtb than Type-A EAE induction, and is NLRP3 inflammasome-independent and ⁇ -resistant.
  • Type-B EAE can also be induced with Type-A EAE induction methods with MHV68 infection or with rl_T ( ⁇ _ ⁇ 2 ⁇ 1) injection.
  • ml_T membrane-bound LT
  • ml_T stimulate LT R on CD4 + T cells, resulting in the upregulation of CXCR2 on CD4 + T cells.
  • Blockade of ⁇ _ ⁇ (with LJ ⁇ R-Fc) and CXCR2 (with SB225002) selectively inhibits the Type-B EAE progression.
  • ml_T is also involved in the induction of Sema6B in T cells. Sema6B causes neural damages, and this may be a reason for the prolonged and minimal remission in Type-B EAE.
  • Provided herein is a method to diagnose subtypes of CNS autoimmune diseases and to more efficiently treat them by determining the subtype.
  • Type B MS Described herein are methods of diagnosing a newly characterized subtype of MS, referred to as Type B MS.
  • Type B MS is non-responsive to the conventional MS treatment with ⁇ .
  • causes for the resistance to ⁇ treatment and the heterogeneity of MS etiology were largely unknown.
  • the inventors have discovered the biochemical distinction between ⁇ -responsive MS and ⁇ - ⁇ -responsive MS, thereby facilitating the classification and diagnosis of Type A MS and Type B MS.
  • Described herein are also methods of treating a central nervous system (CNS) autoimmune disease.
  • the methods may include targeting biomarkers in the biochemical pathways that distinguish Type A MS from Type B MS.
  • CNS central nervous system
  • each intervening number there between with the same degree of precision is explicitly contemplated.
  • the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1 , 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.
  • the term "about” as used herein as applied to one or more values of interest refers to a value that is similar to a stated reference value. In certain aspects, the term “about” refers to a range of values that fall within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11 %, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).
  • administering refers to providing, contacting, and/or delivery of an agent or an inhibitor by any appropriate route to achieve the desired effect.
  • agents or inhibitors may be administered to a subject in numerous ways including, but not limited to, orally, ocularly, nasally, intravenously, topically, as aerosols, suppository, etc. and may be used in combination.
  • biomarker refers to a naturally occurring biological molecule present in a subject at varying concentrations that is useful in identifying and/or classifying a disease or a condition.
  • the biomarker can include genes, proteins,
  • the biomarker comprises a disease marker.
  • the biomarker can be a gene that is upregulated or
  • the biomarker can be a polypeptide whose level is increased or decreased in a subject that has a disease or risk of developing a disease.
  • the biomarker comprises a small molecule.
  • the biomarker comprises a polypeptide.
  • control means "control,” “reference level,” and “reference” are used herein
  • the reference level may be a predetermined value or range, which is employed as a benchmark against which to assess the measured result.
  • Control group refers to a group of control subjects.
  • the predetermined level may be a cutoff value from a control group.
  • the predetermined level may be an average from a control group.
  • Cutoff values (or predetermined cutoff values) may be determined by Adaptive Index Model (AIM) methodology.
  • Cutoff values (or predetermined cutoff values) may be determined by a receiver operating curve (ROC) analysis from biological samples of the patient group.
  • ROC analysis as generally known in the biological arts, is a determination of the ability of a test to discriminate one condition from another, e.g., to determine the performance of each marker in identifying a patient having CRC.
  • cutoff values may be determined by a quartile analysis of biological samples of a patient group.
  • a cutoff value may be determined by selecting a value that corresponds to any value in the 25th-75th percentile range, preferably a value that corresponds to the 25th percentile, the 50th percentile or the 75th percentile, and more preferably the 75th percentile.
  • Such statistical analyses may be performed using any method known in the art and can be implemented through any number of commercially available software packages (e.g., from Analyse-it Software Ltd., Leeds, UK; StataCorp LP, College Station, TX; SAS Institute Inc., Cary, NC).
  • the healthy or normal levels or ranges for a target or for a protein activity may be defined in accordance with standard practice.
  • a control may be a subject, or a sample therefrom, whose disease state is known.
  • the subject, or sample therefrom may be healthy, diseased, diseased prior to treatment, diseased during treatment, diseased after treatment, or healthy after treatment, or a combination thereof.
  • the term "normal subject” as used herein means a healthy subject, i.e.
  • the control is a healthy control.
  • the control comprises Type A MS.
  • the control comprises Type B MS.
  • effective dosage means a dosage of a drug effective for periods of time necessary, to achieve the desired therapeutic result. An effective dosage may be determined by a person skilled in the art and may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the drug to elicit a desired response in the individual.
  • inhibitor or “inhibiting” mean that an activity is decreased or prevented in the presence of an inhibitor as opposed to in the absence of the inhibitor.
  • inhibitor refers to the reduction or down regulation of a process or the elimination of a stimulus for a process, which results in the absence or minimization of the expression or activity of a biomarker or polypeptide. Inhibition may be direct or indirect. Inhibition may be specific, that is, the inhibitor inhibits a biomarker or polypeptide and not others.
  • Polynucleotide as used herein can be single stranded or double stranded, or can contain portions of both double stranded and single stranded sequence.
  • the polynucleotide can be nucleic acid, natural or synthetic, DNA, genomic DNA, cDNA, RNA, or a hybrid, where the polynucleotide can contain combinations of deoxyribo- and ribo-nucleotides, and
  • Polynucleotides can be obtained by chemical synthesis methods or by recombinant methods.
  • a "peptide” or “polypeptide” is a linked sequence of two or more amino acids linked by peptide bonds.
  • the polypeptide can be natural, synthetic, or a modification or combination of natural and synthetic.
  • Peptides and polypeptides include proteins such as binding proteins, receptors, and antibodies.
  • the terms "polypeptide”, “protein,” and “peptide” are used
  • Primary structure refers to the amino acid sequence of a particular peptide.
  • Secondary structure refers to locally ordered, three dimensional structures within a polypeptide. These structures are commonly known as domains, e.g., enzymatic domains, extracellular domains, transmembrane domains, pore domains, and cytoplasmic tail domains. Domains are portions of a polypeptide that form a compact unit of the polypeptide and are typically 15 to 350 amino acids long. Exemplary domains include domains with enzymatic activity or ligand binding activity. Typical domains are made up of sections of lesser organization such as stretches of beta-sheet and alpha-helices.
  • “Tertiary structure” refers to the complete three dimensional structure of a polypeptide monomer. “Quaternary structure” refers to the three dimensional structure formed by the noncovalent association of independent tertiary units.
  • predetermined cutoff and predetermined level as used herein means an assay cutoff value that is used to assess diagnostic, prognostic, or therapeutic efficacy results by comparing the assay results against the predetermined cutoff/level, where the predetermined cutoff/level already has been linked or associated with various clinical parameters (e.g., presence of disease, stage of disease, severity of disease, progression, non-progression, or improvement of disease, etc.).
  • the disclosure provides exemplary predetermined levels.
  • cutoff values may vary depending on the nature of the
  • immunoassay e.g., antibodies employed, reaction conditions, sample purity, etc.
  • reporter capable of generating a detectable signal.
  • the label can produce a signal that is detectable by visual or instrumental means.
  • reporter groups can be used, differing in the physical nature of signal transduction (e.g., fluorescence, electrochemical, nuclear magnetic resonance (NMR), and electron paramagnetic resonance (EPR)) and in the chemical nature of the reporter group.
  • Various reporters include signal-producing substances, such as chromagens, fluorescent compounds, chemiluminescent compounds, radioactive compounds, and the like.
  • the reporter comprises a radiolabel.
  • Reporters may include moieties that produce light, e.g., acridinium compounds, and moieties that produce fluorescence, e.g., fluorescein.
  • the signal from the reporter is a fluorescent signal.
  • the reporter may comprise a fluorophore.
  • fluorophores examples include, but are not limited to, acrylodan (6-acryloy 1-2-dimethylaminonaphthalene), badan (6- bromo-acetyl-2-dimethylamino-naphthalene), rhodamine, naphthalene, danzyl aziridine, 4-[N- [(2-iodoacetoxy)ethyl]-N-methylamino]-7-nitrobenz-2-oxa-1 ,3-diazole ester (IANBDE), 4-[N-[(2- iodoacetoxy)ethyl]-N-methylamino-7-nitrobenz-2-oxa-1 ,3-diazole (IANBDA), fluorescein, dipyrrometheneboron difluoride (BODIPY), 4-nitrobenzo[c][1 ,2,5]oxadiazole (NBD), Alexa fluorescent dyes, and derivatives thereof.
  • acrylodan 6-acryl
  • Fluorescein derivatives may include, for example, 5- fluorescein, 6-carboxyfluorescein, 3'6-carboxyfluorescein, 5(6)-carboxyfluorescein, 6- hexachlorofluorescein, 6-tetrachlorofluorescein, fluorescein, and isothiocyanate.
  • risk assessment means an evaluation of factors including biomarkers, to predict the risk of occurrence of future events including disease onset or disease progression, so that treatment decisions regarding the subject may be made on a more informed basis.
  • Sample or “test sample” as used herein can mean any sample in which the presence and/or level of a biomarker or target is to be detected or determined.
  • Samples may include liquids, solutions, emulsions, mixtures, or suspensions. Samples may include a medical sample.
  • Samples may include any biological fluid or tissue, such as blood, whole blood, fractions of blood such as plasma and serum, peripheral blood mononuclear cells (PBMCs), muscle, interstitial fluid, sweat, saliva, urine, tears, synovial fluid, bone marrow, cerebrospinal fluid, nasal secretions, sputum, amniotic fluid, bronchoalveolar lavage fluid, gastric lavage, emesis, fecal matter, lung tissue, peripheral blood mononuclear cells, total white blood cells, lymph node cells, spleen cells, tonsil cells, cancer cells, tumor cells, bile, digestive fluid, skin, or combinations thereof.
  • the sample comprises an aliquot.
  • the sample comprises a biological fluid.
  • Samples can be obtained by any means known in the art.
  • the sample can be used directly as obtained from a patient or can be pre-treated, such as by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, and the like, to modify the character of the sample in some manner as discussed herein or otherwise as is known in the art.
  • Samples may be obtained before treatment, before diagnosis, during treatment, after treatment, or after diagnosis, or a combination thereof.
  • Subject as used herein can mean a mammal that wants or is in need of the herein described conjugates or fusion proteins.
  • the subject may be a human or a non-human animal.
  • the subject may be a mammal.
  • the mammal may be a primate or a non-primate.
  • the mammal can be a primate such as a human; a non-primate such as, for example, dog, cat, horse, cow, pig, mouse, rat, camel, llama, goat, rabbit, sheep, hamster, and guinea pig; or non- human primate such as, for example, monkey, chimpanzee, gorilla, orangutan, and gibbon.
  • the subject may be of any age or stage of development, such as, for example, an adult, an adolescent, or an infant.
  • beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), whether detectable or undetectable, or enhancement or improvement of the condition, disorder or disease. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.
  • Variant as used herein with respect to a polynucleotide means (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a polynucleotide that is substantially identical to a referenced polynucleotide or the complement thereof; or (iv) a polynucleotide that hybridizes under stringent conditions to the referenced polynucleotide, complement thereof, or a sequences substantially identical thereto.
  • a “variant” can further be defined as a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity.
  • biological activity include the ability to be bound by a specific antibody or polypeptide or to promote an immune response.
  • Variant can mean a substantially identical sequence.
  • Variant can mean a functional fragment thereof.
  • Variant can also mean multiple copies of a polypeptide. The multiple copies can be in tandem or separated by a linker.
  • Variant can also mean a polypeptide with an amino acid sequence that is substantially identical to a referenced polypeptide with an amino acid sequence that retains at least one biological activity.
  • a conservative substitution of an amino acid i.e., replacing an amino acid with a different amino acid of similar properties (e.g., hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes can be identified, in part, by considering the hydropathic index of amino acids. See Kyte et al., J. Mol. Biol. 1982, 157, 105-132. The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydropathic indexes can be substituted and still retain protein function. In one aspect, amino acids having hydropathic indices of ⁇ 2 are substituted.
  • hydrophobicity of amino acids can also be used to reveal substitutions that would result in polypeptides retaining biological function.
  • a consideration of the hydrophilicity of amino acids in the context of a polypeptide permits calculation of the greatest local average hydrophilicity of that polypeptide, a useful measure that has been reported to correlate well with antigenicity and immunogenicity, as discussed in U.S. Patent No. 4,554,101 , which is fully incorporated herein by reference.
  • Substitution of amino acids having similar hydrophilicity values can result in polypeptides retaining biological activity, for example immunogenicity, as is understood in the art.
  • Substitutions can be performed with amino acids having hydrophilicity values within ⁇ 2 of each other.
  • a variant can be a polynucleotide sequence that is substantially identical over the full length of the full gene sequence or a fragment thereof.
  • the polynucleotide sequence can be 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the full length of the gene sequence or a fragment thereof.
  • a variant can be an amino acid sequence that is substantially identical over the full length of the amino acid sequence or fragment thereof.
  • the amino acid sequence can be 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the full length of the amino acid sequence or a fragment thereof.
  • variants include homologues. Homologues may be
  • polynucleotides or polypeptides or genes inherited in two species by a common ancestor are polynucleotides or polypeptides or genes inherited in two species by a common ancestor.
  • CNS disorders affect the structure or function of the spinal cord, the brain, or both, which form the central nervous system.
  • the brain is the main organ of the CNS and is protected by the skull.
  • the spinal cord transmits sensory reception from the peripheral nervous system to the brain, and it also conducts motor information to the body's skeletal muscles, cardiac muscles, smooth muscles, and glands.
  • the spinal cord includes multiple pairs of spinal nerves. These nerves each contain both sensory and motor axons.
  • the spinal cord is protected by vertebrae and connects the peripheral nervous system to the brain.
  • the peripheral nervous system includes the nerves and ganglia beyond the brain and spinal cord.
  • Autoimmune disorders are disorders caused by an abnormal immune response, wherein the immune system attacks the body's own tissues and cells as if they were foreign.
  • CNS autoimmune disorders are those in which the immune system attacks the body's own CNS as if it were foreign.
  • CNS autoimmune diseases include, but are not limited to, multiple sclerosis, myasthenia gravis, Guillain-Barre syndrome, transverse myelitis, and chronic inflammatory demyelinating polyneuropathy. a. Multiple Sclerosis
  • MS Multiple Sclerosis
  • Symptoms of MS include autonomic, visual, motor, and/or sensory problems; difficulty or loss of balance or coordination; numbness; difficulty walking; muscle weakness; bowel and bladder symptoms; eye symptoms including double vision, vision loss, and blindness; muscle spasms; problems with speech; problems with swallowing; acute or chronic pain; depression, mood swings, and memory loss.
  • MS symptoms may be progressive (progressive MS), or they may relapse with symptoms partially or fully improving in between episodes (relapsing-remitting MS, or RRMS).
  • Conventional treatments for MS include, for example, interferon- ⁇ ( ⁇ ), natalizumab, and glatiramer acetate.
  • ⁇ is a conventional first-line treatment for MS, but it is not effective for a significant proportion of MS patients.
  • EAE autoimmune encephalomyelitis
  • the NLRP3 inflammasome plays a role in EAE development. Nlrp3 " ⁇ " mice are resistant to EAE and have fewer Th1 and Th17 cells in the peripheral lymphoid tissues and in the CNS. The lack of the NLRP3 inflammasome in antigen presenting cells (APCs) prevents T helper cells and APCs from migrating to the CNS due to reduced chemokine and chemokine receptor upregulation.
  • APCs antigen presenting cells
  • ⁇ suppresses NLRP3 inflammasome activity; thus, the NLRP3 inflammasome is a target of ⁇ treatment in EAE.
  • EAE can be induced in its absence.
  • EAE induction methods using high dosages of Mtb heat-killed Mycobacteria as an adjuvant
  • Mtb heat-killed Mycobacteria as an adjuvant
  • ⁇ FU$ is effective when EAE is developed in an NLRP3 inflammasome-dependent fashion.
  • I FIS ⁇ -resistant EAE can be induced by altering the intensity of innate immune activation.
  • I FIS ⁇ -resistant EAE subtype As detailed in the Examples, it was found using the EAE model that the addition of extra adjuvant in immunization or acute infection induced an I FIS ⁇ -resistant EAE subtype. Pathogenesis of the I FIS ⁇ -resistant EAE subtype, which involved membrane-bound lymphotoxin and CXCR2 and is characterized by minimal remission, was successfully ameliorated by antagonists of LTfiR or CXCR2. Evaluation of CXCR2 and LTfiR relative mRNA levels identified both I FIS ⁇ -resistant EAE and I FIS ⁇ -nonresponder relapsing-remitting MS patients.
  • mice with IFIS ⁇ -nonresponder relapsing-remitting EAE showed reduced a-motor neuron and shortened neurites mediated by semaphorin 6B on the surface CD4+ T cells. This demonstrated molecular and cellular mechanisms by which the disease heterogeneity is induced.
  • MS may be classified as Type A MS or Type B MS. i) Type A
  • Type A MS refers to MS that is responsive to ⁇ ⁇ treatment.
  • Type A MS may also be referred to as NLRP3 inflammasome-dependent MS.
  • Type A MS may include reversible neuronal damage and demyelination.
  • Type A MS may be characterized by an increased amount of leukocytes, more demyelination, or a combination thereof, in the spinal cord compared to a control.
  • the control is a healthy control.
  • the control comprises Type B MS. ii) Type B
  • Type B MS refers to MS that is non-responsive to I FN ⁇ treatment.
  • Type B MS may also be referred to as N LRP3 inflammasome-independent MS.
  • Subjects with Type-B MS may not display disease remission due to irreversible neuronal damage and demyelination.
  • Type B MS may be characterized by an increased amount of leukocytes, more demyelination, or a combination thereof, in the brain and/or optic nerve compared to a control.
  • Type-B EAE The neuronal damage in Type-B EAE may be caused by a neuronal guidance molecule semaphorin 6B (Sema6B; a class 6 transmembrane semaphorin) expressed on the T cells and upregulated through LT&R signaling.
  • Type B MS may be induced by LT&R signaling and its downstream effectors CXCR2 and Sema6B.
  • Type B MS may also be induced by aggressive immunization. Aggressive immunization may include, for example, administering an antigen such as Mtb in an amount sufficient to induce an acute immune response.
  • LTa lymphotoxin-a
  • DCs dendritic cells
  • Type B MS heterotrimeric ( ⁇ 1 ⁇ 2) membrane-bound lymphotoxin (mLT) on DCs in Type B MS.
  • Type B MS may also induce increased expression of CXCR2 in neutrophils and CD4+ T cells compared to a control.
  • Type B MS is characterized by at least one of irreversible neuronal damage, demyelination, or a combination thereof; irresponsiveness to treatment with ⁇ ; lack of activation of the NLRP3 inflammasome; increased expression of CXCR2, CXCR1 , LT ⁇ R, or Sema6B; increased levels of B cells in the DLNs; or a combination thereof, relative to a control.
  • the control is a healthy control.
  • the expression of CXCR2 is increased in neutrophils, CD4+ cells, or a combination thereof.
  • the control is a healthy control.
  • the control comprises Type A MS.
  • Type B MS may be treated with inhibitors targeting biomarkers that distinguish Type A MS and Type B MS.
  • Type B MS may be treated with inhibitors of LT ⁇ R (mLT receptor), inhibitors of CXCR1 , inhibitors of CXCR2, inhibitors of Sema6, or combinations thereof.
  • Type B MS biomarkers include LT ⁇ R (mLT receptor), CXCR1 , CXCR2, and Sema6.
  • LT ⁇ R mLT receptor
  • CXCR1 CXCR1
  • CXCR2 CXCR2
  • Sema6 Sema6
  • Lymphotoxin beta receptor also known as tumor necrosis factor receptor superfamily member 3 (TNFRSF3), is a cell surface receptor for lymphotoxin.
  • LT ⁇ R specifically binds the lymphotoxin membrane form (mLT), which is a complex of lymphotoxin-alpha (LTa) and lymphtoxin-beta ( ⁇ ). Together with ⁇ , LTa forms the heterotrimeric ( ⁇ 1 ⁇ 2) membrane-bound lymphotoxin (mLT).
  • LT ⁇ R and mLT are involved in apoptosis, cytokine release, and the development and organization of lymphoid tissue and transformed cells.
  • An example of a LT R polypeptide is one according to Accession No.
  • a LT R polypeptide comprises an amino acid sequence of SEQ ID NO: 41 , or a variant thereof.
  • a polynucleotide encodes a polypeptide of SEQ ID NO: 41 , or a variant thereof.
  • Inhibitors of LT R may include antibodies that bind LT&R, siRNA directed to a polynucleotide encoding LT&R, peptides, or small molecule inhibitors specific for LT&R.
  • siRNA directed to a polynucleotide encoding LT&R, peptides, or small molecule inhibitors specific for LT&R.
  • C-X-C motif chemokine receptor 1 (CXCR1 ; also referred to as Interleukin 8 receptor alpha) is a chemokine receptor.
  • CXCR1 binds to Interleukin 8 (IL8) with high affinity and transduces the signal through a G-protein-activated second messenger system. Stimulation of CXCR1 in neutrophils by IL8 leads to neutrophil chemotaxis and activation.
  • IL8 Interleukin 8
  • CXCR1 polypeptide is one according to Accession No. NP_839972.
  • a CXCR1 polypeptide comprises an amino acid sequence of SEQ ID NO: 42, or a variant thereof.
  • a polynucleotide encodes a polypeptide of SEQ ID NO: 42, or a variant thereof.
  • Inhibitors of CXCR1 may include antibodies that bind CXCR1 , siRNA directed to a polynucleotide encoding CXCR1 , peptides, or small molecule inhibitors specific for CXCR1.
  • CXCR2 antibodies that bind CXCR1 , siRNA directed to a polynucleotide encoding CXCR1 , peptides, or small molecule inhibitors specific for CXCR1.
  • C-X-C motif chemokine receptor 2 (CXCR2; also referred to as Interleukin 8 receptor beta) is a chemokine receptor.
  • CXCR2 binds to Interleukin 8 (IL8) with high affinity and transduces the signal through a G-protein-activated second messenger system.
  • CXCR2 also binds CXCL1 , CXCL2, CXCL3, and CXCL5.
  • CXCR2 polypeptide is one according to Accession No. NP_034039.
  • a CXCR2 polypeptide comprises an amino acid sequence of SEQ ID NO: 43, or a variant thereof.
  • a polynucleotide encodes a polypeptide of SEQ ID NO: 43, or a variant thereof.
  • Inhibitors of CXCR2 may include antibodies that bind CXCR2, siRNA directed to a polynucleotide encoding CXCR2, peptides, or small molecule inhibitors specific for CXCR2.
  • the CXCR2 inhibitor comprises SB225002 (N-(2-Bromophenyl)-N'-(2- hydroxy-4-nitrophenyl)urea, which may be purchased from Tocris (Bristol, UK), catalog no. 2725).
  • SB225002 N-(2-Bromophenyl)-N'-(2- hydroxy-4-nitrophenyl)urea, which may be purchased from Tocris (Bristol, UK), catalog no. 2725).
  • Semaphorin 6B is a class 6 transmembrane semaphorin. Semaphorins are a class of secreted and membrane proteins that were originally identified as axonal growth cone guidance molecules. Semaphorins may signal through multimeric receptor complexes and may act as short-range inhibitory signals. Semaphorins may act as cues to deflect axons from inappropriate regions, as in, for example, neural system development.
  • Sema6B polypeptide is one according to Accession No.
  • a Sema6B polypeptide comprises an amino acid sequence of SEQ ID NO: 44, or a variant thereof.
  • a polynucleotide encodes a polypeptide of SEQ ID NO: 44, or a variant thereof.
  • Inhibitors of Sema6 may include antibodies that bind Sema6, siRNA directed to a polynucleotide encoding Sema6, peptides, or small molecule inhibitors specific for Sema6.
  • the term “detect” or “determine the presence of” refers to the qualitative measurement of undetectable, low, normal, or high concentrations of one or more biomarkers. Detection may include in vitro, ex vivo, or in vivo detection. Detection may include detecting the presence of one or more biomarkers versus the absence of the one or more biomarkers. Detection may also include quantification of the level of one or more biomarkers. The term “quantify” or “quantification” may be used interchangeably, and may refer to a process of determining the quantity or abundance of a substance (e.g., biomarker), whether relative or absolute.
  • a substance e.g., biomarker
  • Biomarkers or expression thereof as described herein may be detected and/or measured at the polynucleotide level, the polypeptide level, or a combination thereof.
  • Expression may be detected and/or measured by any means known by one of skill in the art such as, for example, immunohistochemistry, antibody binding, hybridization of a probe, arrays, microarrays, PCR, real time RT-PCR, RNA in situ hybridization, RNAse protection assay, Northern analysis, magnetic particles (e.g., microparticles or nanoparticles), Southern analysis, Western analysis, ELISA, expression reporter plasmids, DNA-chip analysis with primers, HPLC, mass spectrometry, protein microarray analysis, PAGE analysis, isoelectric focusing, 2-D gel electrophoresis, enzymatic assays, and any method or system involving flow cytometry.
  • any means known by one of skill in the art such as, for example, immunohistochemistry, antibody binding, hybridization of a probe, arrays, microarrays, PCR, real time RT-PCR, RNA in situ hybridization, RNAse protection assay, Northern analysis, magnetic particles (e.g., microparticle
  • Binding with antibodies may include, for example, antibodies tethered to or associated with a reporter or an imaging agent, ELISA, and arrays.
  • the ELISA may be a sandwich ELISA.
  • the biomarker itself as a polypeptide is detected by antibody binding or any suitable immunohistochemistry method.
  • a polynucleotide encoding the biomarker may be detected by hybridization with a probe.
  • expression of a polynucleotide encoding the biomarker may be detected using microarray analysis.
  • an antibody or probe may be labeled with a reported for detection.
  • the amount or level of polypeptide or gene expression may be quantified.
  • expression may be measured by determining at least one of mRNA levels, protein levels, or protein activity levels of the biomarker.
  • a biomarker or expression thereof is detected at the polynucleotide level.
  • at least one nucleic acid probe specifically binds or hybridizes to a polynucleotide encoding the biomarker polypeptide to form a complex, and the complex is detected.
  • RT-PCR is used to amplify a polynucleotide encoding the biomarker polypeptide with specific primers.
  • quantitative RT-PCR is used to amplify a polynucleotide encoding the biomarker polypeptide with specific primers.
  • rolling circle amplification is used to amplify a polynucleotide encoding the biomarker polypeptide.
  • strand displacement amplification is used to amplify a polynucleotide encoding the biomarker polypeptide.
  • a biomarker or expression thereof is detected at the polypeptide level.
  • a biomarker or expression thereof is detected by using molecules which bind to the biomarker polypeptide. Suitable molecules/agents which bind either directly or indirectly to the biomarker polypeptide in order to detect its presence include naturally occurring molecules such as peptides and proteins, for example antibodies, or they may be synthetic molecules.
  • a sample is contacted with an antibody capable of binding the biomarker polypeptide and monitoring the sample for the presence of the polypeptide.
  • the antibody may be a monoclonal antibody or polyclonal antibody.
  • the complex formed between the antibody and the biomarker polypeptide may be monitored by any method known in the art, such as, for example, fluorescence resonance energy transfer (FRET), and surface plasmon resonance (SPR).
  • FRET fluorescence resonance energy transfer
  • SPR surface plasmon resonance
  • the biomarker polypeptide is detected by an immunoassay.
  • Immunoassays may include, for example, competitive and noncompetitive assay systems, Western analysis, radioimmunoassays, ELISA, sandwich immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, and protein A immunoassays.
  • Specific immunological binding of the antibody to the biomarker polypeptide can be detected via direct labels, attached to the antibody or via indirect labels, such as alkaline phosphatase or horseradish peroxidase.
  • the use of immobilized antibodies may be incorporated into the immunoassay.
  • the antibodies may be immobilized onto a variety of supports, such as magnetic or chromatographic matrix particles, the surface of an assay plate (such as microtiter wells), pieces of a solid substrate material, and the like.
  • An assay strip can be prepared by coating the antibody or plurality of antibodies in an array on a solid support. This strip can then be dipped into the test biological sample and then processed quickly through washes and detection steps to generate a measurable signal, such as a colored spot.
  • a composition may comprise the inhibitor.
  • the inhibitors of biomarkers as detailed above can be formulated into a composition in accordance with standard techniques well known to those skilled in the pharmaceutical art.
  • the composition may be prepared for administration to a subject.
  • Such compositions can be administered in dosages and by techniques well known to those skilled in the medical arts taking into consideration such factors as the age, sex, weight, and condition of the particular subject, and the route of administration.
  • the inhibitor can be administered prophylactically or therapeutically.
  • the inhibitor can be administered in an amount sufficient to induce a response.
  • the inhibitors are administered to a subject in need thereof in an amount sufficient to elicit a therapeutic effect.
  • An amount adequate to accomplish this is defined as "therapeutically effective dose.” Amounts effective for this use will depend on, e.g., the particular composition of the inhibitor regimen administered, the manner of administration, the stage and severity of the disease, the general state of health of the patient, and the judgment of the prescribing physician.
  • the inhibitor can be administered by methods well known in the art as described in Donnelly et al. (Ann. Rev. Immunol. 1997, 15, 617-648); Feigner et al. (U.S. Patent No. 5,580,859, issued Dec. 3, 1996); Feigner (U.S. Patent No. 5,703,055, issued Dec. 30, 1997); and Carson et al. (U.S. Patent No. 5,679,647, issued Oct. 21 , 1997), the contents of all of which are incorporated herein by reference in their entirety.
  • the inhibitor can be complexed to particles or beads that can be administered to an individual, for example, using a vaccine gun.
  • a pharmaceutically acceptable carrier including a physiologically acceptable compound, depends, for example, on the route of administration.
  • the inhibitor can be delivered via a variety of routes. Typical delivery routes include parenteral administration, e.g., intradermal, intramuscular or subcutaneous delivery. Other routes include oral administration, intranasal, intravaginal, transdermal, intravenous,
  • the inhibitor is administered intravenously, intraarterially, or intraperitoneally to the subject.
  • the inhibitor can be a liquid preparation such as a suspension, syrup, or elixir.
  • the inhibitor can be incorporated into liposomes, microspheres, or other polymer matrices (such as by a method described in Feigner et al., U.S. Patent No. 5,703,055; Gregoriadis, Liposome Technology, Vols. I to III (2nd ed. 1993), the contents of which are incorporated herein by reference in their entirety).
  • Liposomes can consist of phospholipids or other lipids, and can be nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.
  • the inhibitor may be used as a vaccine.
  • the vaccine can be administered via electroporation, such as by a method described in U.S. Patent No. 7,664,545, the contents of which are incorporated herein by reference.
  • the electroporation can be by a method and/or apparatus described in U.S. Patent Nos. 6,302,874; 5,676,646; 6,241 ,701 ; 6,233,482;
  • the electroporation can be carried out via a minimally invasive device.
  • the inhibitor is administered in a controlled release formulation.
  • the inhibitor may be released into the circulation, for example.
  • the inhibitor may be released over a period of at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 1 week, at least about 1.5 weeks, at least about 2 weeks, at least about 2.5 weeks, at least about 3.5 weeks, at least about 4 weeks, or at least about 1 month.
  • CNS central nervous system
  • the method may include administering to the subject at least one of an LT ⁇ R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • the methods may include determining the expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in a sample from the subject, wherein an increase of the expression relative to a control indicates a diagnosis that the subject is afflicted with Type B MS.
  • the methods further include diagnosing the patient as having Type B MS when the expression is increased relative to the control.
  • the control is a healthy control.
  • the control comprises Type A MS.
  • the methods further include administering treatment with one or more
  • compositions to the subject diagnosed as having Type B MS.
  • the methods may include (a) contacting a sample from the subject with an agent for specifically detecting expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (b) determining a level of expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; and (c) comparing the level of expression in the sample with a control level of expression to diagnose Type B MS.
  • the methods further include diagnosing the patient as having Type B MS when the level of expression is increased relative to the control.
  • the control is a healthy control.
  • the control comprises Type A MS.
  • the methods further include administering treatment with one or more pharmaceutical compositions, to the subject diagnosed as having Type B MS. c.
  • the methods may include (a) contacting a sample from the subject with an agent for specifically detecting expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (b) determining a level of expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (c) comparing the level of expression in the sample with a control level of expression; and (d) determining that the subject is not responsive to treatment with ⁇ when the level of expression is increased relative to the control, and that the subject is responsive to treatment with ⁇ when the level of expression is the same or decreased relative to the control.
  • the methods further include administering treatment with ⁇ , to the subject determined to be responsive to treatment with ⁇ . In some embodiments, the methods further include administering treatment to the subject determined to be not responsive to treatment with ⁇ , wherein the treatment includes at least one of an LT R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • the sample is obtained before treatment. In some embodiments, the sample is obtained during treatment. In some embodiments, the sample is obtained after treatment. In some embodiments, the sample is obtained before, during, and/or after treatment.
  • the control is a healthy control. In some embodiments, the control comprises Type A MS. In some embodiments, the control is a sample from the subject obtained prior to treatment. In some embodiments, the control is a sample from the subject obtained during treatment. In some embodiments, the control is a sample from the subject obtained after treatment.
  • Cxcr2 fl fl mice were a gift from Dr. Richard M. Ransohoff (Cleveland Clinic)(Liu et al. Genesis 2013, 51 , 587- 595).
  • Lck-Cre transgenic mice were purchased from Jackson Laboratories.
  • C57BU6 mice were purchased from Jackson Laboratories. All the mice were kept in a specific pathogen free facility. This study was approved by the Duke University Institutional Animal Care and Use Committee.
  • MOG 3 5.55 peptide was emulsified with CFA (including Mtb, heat- killed Mycobacteria) and subcutaneously injected in the flanks of mice on day 0 for one-time immunization, or day 0 and 7 for two-time immunization. Amounts of MOG peptide and Mtb are described in FIG. 8. In addition to whether immunization was performed once or twice,
  • Pertussis toxin (PTx; 200 ng/mouse) was administered on day 0 and 2, if the MOG
  • Antibodies against LT (ab100844), CXCR2 (129101), and CXCL1 (LS-B2513) were purchased from Abeam, Biolegend, and Life Span Bioscience, respectively.
  • the ⁇ used in EAE treatment was human r!FNp- 1 b (Betaseron, Bayer).
  • rLT ( ⁇ 2 ⁇ 1 ; 1008-LY-010/CF) was purchased from R & D systems.
  • Mouse rCXCLI (573702), rlL-17 (576002), and rIFNv (575306) were purchased from Biolegend.
  • MOG 35-55 peptide was synthesized by United Peptides.
  • Enzyme-linked immunosorbent assay (EUSA) kits for the detection of ⁇ L- f ⁇ and CXCL1 were purchased from BD PharMingen, and Promocel!, respectively.
  • MHV68 EAE induction by Type-A EAE with MHV68 infection.
  • MHV68 was purchased from ATCC Ceil lines (VR-1465).
  • MHV68 strains were amplified in BALB/3 TI2-3 ceils (ATCC, CCL-164TM), and virus-infected cells were treated with three freeze-thaw cycles, and then supernatant was isolated.
  • Titers of purified virus were determined using the median tissue culture infective dose (TCID50) method, and convert TCID50 to plaque forming units (PFU).
  • TCID50 median tissue culture infective dose
  • PFU plaque forming units
  • mice with EAE ⁇ (3x10 4 unit/mouse) were i.p. injected every other day from day 0 to 8 as previously performed (Inoue et al. Sci. Signal 2012, 5, ra38).
  • rLT ⁇ 32 ⁇ 1(1008- ⁇ -0 0/ ⁇ , R&D; 10 Mg/kg mouse) was i.p. administered every day from day 0 to 9.
  • CXCR2 inhibitor SB225002, Tocris: 0.1 pg/kg mouse was i.p. injected every day from day 0 to day 20.
  • I ⁇ R-Fc PR0154527, Genentech
  • CD4+ T cells were isolated with CD4 microbead (Myltenyi) from the spleen of naive mice.
  • DCs were isolated with CD1 1c microbead (Myltenyi) from DLNs of naive mice in all the experiments.
  • DCs were stimulated with riL-17 (10 ng/mL) or r!FNy (10 ng/mL) for 3h.
  • Total RNA isolation, cDNA synthesis and qPCR were performed as described below.
  • RNA and cDNA preparation and qPCR and PCR array analyses were isolated by FACS-sorting on 9-dpi. Total RNA was extracted from cells with RNeasy Kit (Qiagen). cDNA synthesis was performed with qScript cDNA SuperMix (Quanta).
  • qPCR quantitative polymerase chain reaction
  • PCR array was performed by using Chemokines & Receptors PGR Array (PA M-022Z, Qiagen) and Cytokines & Chemokines PGR Array (PA M-150Z, Qiagen) with CD4 ⁇ T cells isolated from spleen and DCs isolated from DLNs at 9-dpi, respectively.
  • RNA-seq analysis For RNA-seq analysis, CD3 ⁇ CD4+ cells (as CD4+ T cells) were purified by FACS-sorting from spleens of two mice at 9-dpi. Total RNA was extracted from cells with RNeasy Kit (Qiagen). Stranded mRNA libraries were made with the !!iumina TruSeq Stranded mRNA kit (catalog number RS-122-2101). Before library preparation, the RNA quality was evaluated using a TapeStation instrument (Agilent technologies) and using a Qubit (Life Technologies). The quality of RNA libraries was checked by running them on a 2100
  • Bioanalyzer instrument (Agilent), pooled and sequenced on 2 lanes of lliumina HiSeq 50bp SR. RNA-seq data was processed using the TrimGa!ore toolkit
  • NCBI .38r73 version of the mouse genome and transcriptome (Kersey et al. Nucleic Acids Research 2012, 40, D91-97) using the STAR RNA-seq alignment tool (Dobin et al.
  • RRMS patient's information and qPCR analysis of PBMC samples from RRMS patients were obtained from the MURDOCH study cohorts (www.murdock-study.com). Each participant self-reported their drug history, which included the drug, start and end dates, and comments. Comments on discontinued drugs typically included the reason for discontinuation, and we selected group 2 individuals (non-responders) as those that self-reported stopping an interferon drug due to it not working. Relapses and more lesions via RI were often cited, and some referred to positive neutralizing antibody results, but sometimes they just reported that it was not working without further description. Group 1 individuals (responders) were currently taking an interferon drug with no reports of disease progression, and we weighted selection to those that explicitly reported that the drug was working for them and that they had not had relapses or developed more lesions while on it.
  • RNA from samples responder and non-responder groups were quantified via
  • DCs were FACS-sorted as CD11 c+CD1 1 b- from DLNs of naive and Type-A and Type-B EAE mice at 9-dpi. Genomic DNA was obtained with the
  • the perfused mouse brains were kept within the cranium to prevent any potential damage to the brain caused by surgical removal.
  • Ail images were acquired using a 9.4 T (400 Hz) 89-mm vertical bore Oxford magnet with shielded coil providing gradients of 2200 mT/m.
  • the system was controlled by the Agilent imaging console.
  • the specimen was positioned tightly inside a cylindrical polyethylene tube which was filled with Fombiin (perfiuoropolyether; Ausimont, Inc., Morristown, NJ) that did not emit proton MR signal thus providing dark background in the image.
  • the tube was sealed with care to avoid air bubbles entering the tube.
  • Specimens were imaged in a solenoid radiofrequency coil constructed from a single sheet of microwave substrate.
  • the diameter of the solenoid coil was closely matched to the specimen container thus providing high signal sensitivity.
  • FLAIR 2D fluid attenuated inversion recovery
  • Neuronal staining and motor neuron analysis by stereoiogy Neuronal staining was performed by Goigi's silver staining with the FD Rapid Goigi staining kit according manufacture's protocol (FD Neurotechonologies, inc.).
  • FD Neurotechonologies, inc. FD Neurotechonologies, inc.
  • EAE mice at 70 dpi were euthanized by C02 and transcardially perfused with PBS followed by cold 4% paraformaldehyde in 0.1 M phosphate buffer. Fixed spinal cords were removed and
  • the dissector height was 18 m and the counting frame size was 75 x 75 ⁇ Stereoiogy parameters were indicated in TABLE 4.
  • the total numbers of bilateral Nissl- stained ventral horn neurons were counted stereoiogicaily with Stereo Investigator software by using an optical fractionator.
  • a-motor neurons were defined by having a maximal diameter greater than 30 m.
  • Total neuron numbers included a-motor neurons and Nissl-stained neurons having a diameter less than 30 ⁇ to ensure that a change in motor neuron number did not reflect changes in cell size.
  • the average number of a-motor neurons (>30 ⁇ ) was 162.
  • the average number of total Nissl-stained neurons counted per animal was 337.
  • CD4 ⁇ T cell-neuron co-culture, and neurite length measurement Primary neurons were obtained from the hippocampus of P2 mice, as described previously (Banker and Goslin Nature 1988, 336, 185-186). After papain treatment to hippocampus tissue, 1x10 s cells were plated on poly-L lysine-coated 8 mm diameter glass slides and were cultured for 8 days. CD4+ T cells were isolated with CD4+ microbead ( yltenyi) from the spleen of naive mice and EAE at 9-dpi, and cultured with primary cultured neuron for 24 h. Neuron and nuclear staining were performed with MAP2 and DAPL respectively.
  • Hot-plate test To evaluate thermal sensitivity, hot plate test (Eddy and Leimbach The Journal of Pharmacology and Experimental Therapeutics 1953, 107, 385-393) was used for mice at 9-dpi. The mice were yet to show any EAE symptoms and motor dysfunction. The latency to nociceptive responses, such as licking of paw or flicks hind paw, were measured, when they are placed on a warmed metal plate at constant temperature (52°C). A cut-off time of40 sec was used to avoid pain and tissue injury as much as possible. [000104] Chemotaxis assay.
  • CD4 ⁇ T cells were obtained from naive mice or mice with either Type-A EAE or Type-B EAE at 9-dpi, and submitted for chemotaxis assays as previously described (Inoue et ai. PNAS 2012109, 10480-10485). Briefly, CD4 ⁇ T cells (1x10 6 cells/well) were resuspended in the RPMI 1640 medium containing 5% FBS, and plated in upper chambers of Transweil (5 pm pore, Coming Costar). rCXCL.1 was added to lower chambers, and cells were incubated for 3 hr at 37°C. Numbers of migrated cell were counted, and the data are displayed after subtracting ceil numbers in lower chambers in untreated group (as control),
  • Type-B EAE did not ameliorate Type-B EAE both in WT and Nlrp3-1 mice (FIG. 1 F; FIG. 8D), while Type-A EAE responded well to ⁇ treatment (FIG. 1 F, right panel). Severity of Type-B EAE was similar to Type-A EAE, whereas ⁇ treatment readily distinguished these two types of disease.
  • Type-B EAE is induced with high Mtb dosages, i.e., high levels of microbial adjuvant, used in EAE immunization (FIG. 8)
  • MHV-68 murine gamma herpesvirus-68
  • EBV infection is one of the risk factors for MS.
  • Type-B EAE mice showed significantly higher numbers of various leukocytes in brains than Type-A EAE mice on 17-dpi (peak time of disease)(FIG. 1 H, FIG. 8E). In contrast, Type-A EAE mice showed higher numbers of these cells in spinal cords than Type-B EAE mice (FIG. 1 H, FIG. 8E). The data strongly suggest that Type-B EAE tends to attract immune cells to the brain.
  • Type-A EAE mice suffered severe demyelination in the spinal cord, whereas this demyelination was milder in Type-B EAE mice (FIG. 11).
  • Type-B EAE mice but not Type-A EAE mice, suffered severe demyelination in the brain (proximal to choroid plexus, an entry zone of immune cells into the brain) and in the optic nerve (FIG. 11).
  • MRI T2 FLAIR images also showed hyperintensity in the external capsule and adjacent cortical regions in the brains of Type-B EAE mice at 18-dpi and to a lesser extent in the spinal cord (FIG. 1 J, FIG. 8F). The results indicate possible loss of myelin in the brain cortical region and milder loss in the spinal cord in Type-B EAE mice.
  • Type-B EAE does not require the NLRP3 inflammasome for the disease development (FIG. 1), we hypothesized that an as-yet-unknown molecular pathway is involved.
  • PCR array analyses indicated high expression of Lta mRNAs in DCs isolated from draining lymph nodes (DLNs) of Type-B EAE mice compared to Type-A EAE mice (TABLE 1).
  • Lta encodes lymphotoxin a (LTa, also termed ⁇ ), which forms secreted homotrimer or membrane-bound hetero-trimer with ⁇ (mLT as membrane LT).
  • LT is known to be critical in the development of MS and EAE.
  • LT was observed in demyelinated regions in the brain of some MS patient. Serum LT levels were reported to be elevated in a subgroup of MS patients with high numbers of lesions and disease burden.
  • Blockade of LTfiR, an mLT receptor ameliorates EAE induced by repetitive immunizations, a condition that approximates Type-B EAE induction.
  • Lta mRNA and mLT expression in DCs from Type-B EAE mice, but not from naive and Type-A EAE mice (FIG. 2A, FIG. 2B; FIG. 9A), suggesting involvement of mLT in Type-B EAE development.
  • Th17-mediated passive EAE did not ameliorate EAE induced by Th17 cell adoptive transfer. Therefore, we sought a connection between Th17-mediated passive EAE and Type-B EAE.
  • Th17 cells may induce Type-B EAE disease.
  • LT ⁇ R-Fc strongly ameliorated Th17-mediated passive EAE (FIG. 2I).
  • Th17 cell transfer made Asc "A recipients develop severe EAE (FIG. 2I), although MOG-stimulated unpolarized T cells were not potent enough to drive EAE in recipients lacking the NLRP3 inflammasome (Inoue et al. Sci. Signal 2012, 5, ra38).
  • Type-B EAE [000120] Interestingly, several pattern-recognition receptors and neuronal damage-related molecules are upregulated in Type-B EAE (FIG. 3C), suggesting enhanced inflammation and neuronal damages in Type-B EAE. The results strongly suggested Type-A and Type-B EAE have quite distinct gene expression profiles in splenic CD4+ T cells.
  • CXCR2 is generally known as a receptor on neutrophils, but more than 10% of CD4+ T cells from Type-B EAE mice expressed CXCR2 on their cell surface (FIG. 40, FIG. 11 D).
  • MFI mean fluorescent intensity
  • CXCR2+ macrophages were also more frequent and expressed higher levels of CXCR2 in Type-B EAE, although the CXCR2+ macrophage population was small (FIG. 11 E).
  • mRNA expression levels of Cxcr2, Cxcrl , and Ltbr may be used to distinguish an IFIS ⁇ -resistant subtype.
  • PBMCs peripheral blood mononuclear cells
  • FIG. 5B Blocking LTfiR in Type-B EAE mice reduced CXCL1 levels both in serum and choroid plexus (FIG. 5D). Therefore, enhancement of the CXCL1 -CXCR2 interaction is a key effector response downstream of LT in the development of Type-B EAE. Indeed, blockade of CXCR2 ameliorated Type-B EAE, but not Type-A EAE (FIG. 5E).
  • CXCR2 cKO conditional knockout of CXCR2
  • Type-A EAE was developed similarly between WT and CXCR2 cKO mice, but CXCR2 cKO mice were significantly protected from the induction of Type-B EAE in an early phase (FIG. 5F).
  • CXCR2 cKO mice eventually catch up with WT mice in EAE severity later and this suggest the involvement of CXCR2 on other cells, such as neutrophil and macrophages in Type-B EAE.
  • CXCR1/2 On T cells and enhanced expression of CXCL1 in an LT mediated fashion in Type-B EAE.
  • Type-B EAE mice showed persistent EAE severity with little remission (FIG. 6A).
  • Type-B EAE mice displayed severe myelin loss in the spinal cord at 60-dpi as the disease did not remit (FIG. 6B).
  • spinal cords in Type-B EAE mice at 70-dpi were devoid of cell infiltration in the CNS as well as those in Type-A EAE mice (FIG. 13A). Then, we considered possibility of poor remyelination in Type-B EAE by evaluating insulin-like growth factor (IGF)-I, which plays a key role in
  • mice with both subtypes of EAE showed similarly increased levels of IGF-1 in the spinal cord on 22-dpi, compared with WT control (FIG. 13B), suggesting that at least IGF-1-mediated remyelination may not be devoid in Type-B EAE.
  • Type-A EAE mice showed similar neuronal morphology and number of a-motor neurons to naive mice (FIG. 6C, FIG. 6D, FIG. 13C, FIG. 13D). Based on the minimal remission in Type-B EAE, it is possible that axon retraction played a role in making Type-B EAE severity irreversible.
  • Example 10
  • T cells in Type-B EAE mice induce neuronal retraction by Sema6B
  • a method of treating a central nervous system (CNS) autoimmune disease in a subject comprising administering to the subject at least one of an LT R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • CNS central nervous system
  • Clause 2 The method of clause 1 , wherein the inhibitor comprises siRNA, an antibody, or a small molecule.
  • Clause 10 The method of clause 9, wherein expression of CXCR2 is increased in neutrophils, CD4+ cells, or a combination thereof.
  • Clause 1 A method of diagnosing Type B MS in a subject, the method comprising: determining the expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in a sample from the subject, wherein an increase of the expression relative to a control indicates a diagnosis that the subject is afflicted with Type B MS.
  • Clause 12 The method of clause 11 , further comprising diagnosing the patient as having Type B MS when the expression is increased relative to the control.
  • Clause 13 The method of clause 11 , further comprising administering treatment with one or more pharmaceutical compositions, to the subject diagnosed as having Type B MS.
  • Clause 14 The method of clause 13, wherein the treatment comprises at least one of an LT ⁇ R inhibitor, a CXCR2 inhibitor, a CXCR1 inhibitor, or a Sema6B inhibitor, or a combination thereof.
  • a method of diagnosing Type B MS in a subject comprising: (a) contacting a sample from the subject with an agent for specifically detecting expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (b) determining a level of expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; and (c) comparing the level of expression in the sample with a control level of expression to diagnose Type B MS.
  • Clause 16 The method of clause 15, wherein the agent comprises a polynucleotide probe that hybridizes specifically with RNA transcribed from gene encoding at least one of CXCR2, CXCR1 , Ltbr, and Sema6B present in the subject.
  • Clause 17 The method of clause 15, wherein the agent comprises an antibody that binds to at least one of CXCR2, CXCR1 , Ltbr, and Sema6B.
  • Clause 18 The method of any one of clauses 15-17, further comprising diagnosing the patient as having Type B MS when the level of expression is increased relative to the control.
  • a method of predicting responsiveness of a subject with MS to treatment with ⁇ comprising: (a) contacting a sample from the subject with an agent for specifically detecting expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (b) determining a level of expression of at least one of CXCR2, CXCR1 , Ltbr, and Sema6B in the sample; (c) comparing the level of expression in the sample with a control level of expression; and (d) determining that the subject is not responsive to treatment with ⁇ when the level of expression is increased, and that the subject is responsive to treatment with ⁇ when the level of expression is the same or decreased.
  • Clause 21 The method of any one of clauses 9-20, wherein the expression is measured by determining at least one of mRNA levels, protein levels, or protein activity levels.
  • Clause 22 The method of any one of clauses 1 1-21 , wherein the determining step comprises at least one of PCR or antibody binding.
  • Sema6B from Mus musculus Accession No. NP_001123928, 886 amino acids

Abstract

L'invention concerne des compositions et des méthodes pour le diagnostic et le traitement de maladies autoimmunes du SNC telles que la sclérose en plaques. Les méthodes de traitement peuvent comprendre l'administration au patient d'au moins l'un des éléments suivants : un inhibiteur de LTβR, un inhibiteur de CXCR2, un inhibiteur de CXCR1, ou un inhibiteur de Sema6B, ou une association de ces derniers. La méthode permettant de diagnostiquer une sclérose en plaque de type B peut comprendre l'évaluation, dans un échantillon provenant du patient, de l'expression d'au moins l'un des éléments suivants : CXCR2, CXCR1, Ltbr et Sema6B, une augmentation de l'expression en comparaison à un contrôle indiquant un diagnostic de sclérose en plaque de type B chez le patient.
PCT/US2016/058285 2015-10-22 2016-10-21 Méthodes et compositions pour le diagnostic et le traitement de maladies autoimmunes du système nerveux central WO2017070589A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201562244885P 2015-10-22 2015-10-22
US62/244,885 2015-10-22

Publications (1)

Publication Number Publication Date
WO2017070589A1 true WO2017070589A1 (fr) 2017-04-27

Family

ID=58558204

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2016/058285 WO2017070589A1 (fr) 2015-10-22 2016-10-21 Méthodes et compositions pour le diagnostic et le traitement de maladies autoimmunes du système nerveux central

Country Status (1)

Country Link
WO (1) WO2017070589A1 (fr)

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060003327A1 (en) * 2002-03-21 2006-01-05 Anat Achiron Peripheral blood cell markers useful for diagnosing multiple sclerosis and methods and kits utilizing same
US20100256157A1 (en) * 2002-10-29 2010-10-07 Jakob Busch-Petersen Method of treatment
US20110177094A1 (en) * 2007-08-13 2011-07-21 Baxter International Inc. Ivig modulation of chemokines for treatment of multiple sclerosis, alzheimer's disease, and parkinson's disease
US20120046243A1 (en) * 2010-08-23 2012-02-23 Syntrix Biosystems Inc. Aminopyridine- and Aminopyrimidinecarboxamides as CXCR2 Modulators
US20120177632A1 (en) * 2011-01-10 2012-07-12 Shinohara Mari L Methods of optimizing disease treatment
WO2014127258A2 (fr) * 2013-02-14 2014-08-21 Cornell University Méthodes de protection contre la sclérose en plaques et méthodes pour la traiter
WO2014170317A1 (fr) * 2013-04-17 2014-10-23 Morphosys Ag Anticorps ciblant spécifiquement le cxcr2 humain

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060003327A1 (en) * 2002-03-21 2006-01-05 Anat Achiron Peripheral blood cell markers useful for diagnosing multiple sclerosis and methods and kits utilizing same
US20100256157A1 (en) * 2002-10-29 2010-10-07 Jakob Busch-Petersen Method of treatment
US20110177094A1 (en) * 2007-08-13 2011-07-21 Baxter International Inc. Ivig modulation of chemokines for treatment of multiple sclerosis, alzheimer's disease, and parkinson's disease
US20120046243A1 (en) * 2010-08-23 2012-02-23 Syntrix Biosystems Inc. Aminopyridine- and Aminopyrimidinecarboxamides as CXCR2 Modulators
US20120177632A1 (en) * 2011-01-10 2012-07-12 Shinohara Mari L Methods of optimizing disease treatment
WO2014127258A2 (fr) * 2013-02-14 2014-08-21 Cornell University Méthodes de protection contre la sclérose en plaques et méthodes pour la traiter
WO2014170317A1 (fr) * 2013-04-17 2014-10-23 Morphosys Ag Anticorps ciblant spécifiquement le cxcr2 humain

Similar Documents

Publication Publication Date Title
Li et al. Delayed microglial depletion after spinal cord injury reduces chronic inflammation and neurodegeneration in the brain and improves neurological recovery in male mice
Shin et al. Macrophage migration inhibitory factor regulates U1 small nuclear RNP immune complex–mediated activation of the NLRP3 Inflammasome
Inoue et al. An interferon-β-resistant and NLRP3 inflammasome–independent subtype of EAE with neuronal damage
US11136626B2 (en) Biomarkers for the diagnosis of lacunar stroke
JP5555626B2 (ja) 多発性硬化症、アルツハイマー病およびパーキンソン病を処置するためのケモカインのｉｖｉｇ調節
JP5650871B1 (ja) そう痒を伴う疾患に罹患した患者のｉｌ−３１アンタゴニストによる治療に対する応答を予測する方法
JP2010535825A5 (fr)
US20120177632A1 (en) Methods of optimizing disease treatment
KR20110014979A (ko) 항바이러스 요법
JP2014513289A (ja) ＩＦＮβに対する治療応答性を予測するバイオマーカーおよびその使用
KR20210084528A (ko) I형 인터페론 시그너처 및 사용 방법
US7709215B2 (en) Method for diagnosing and treating acute joint injury
BRPI0618609A2 (pt) biomarcadores para tratamento com anticorpo anti-nogo-a em danos à medula espinhal
WO2016042114A1 (fr) Cxcl14 en tant que biomarqueur de l'activité de la voie hedgehog pour diagnostic, le pronostic et le traitement d'une fibrose pulmonaire idiopathique
JP2013213774A (ja) 結核検査用バイオマーカー
WO2017070589A1 (fr) Méthodes et compositions pour le diagnostic et le traitement de maladies autoimmunes du système nerveux central
WO2014038622A1 (fr) Biomarqueur pour maladies autoimmunes
CN110740733A (zh) Irak4介导的疾患和病症的诊断和治疗方法
Hamed et al. Association of interleukin-6 and its-174G/C promoter polymorphism with clinical and laboratory characteristics of non hepatitis C virus rheumatoid arthritis patients
US9945843B2 (en) Methods for identifying compounds that inhibit G protein-coupled receptor (GPR84) agonist-stimulated chemotaxis
KR20140109956A (ko) 테나신-c 및 류마티스 관절염에서의 이의 용도
Inoue et al. Mechanism to develop inflammasome-independent and interferon-β-resistant EAE with neuronal damages
KR102325972B1 (ko) 간암 치료 반응 예측용 spp1 바이오마커 및 이의 용도
KR102494928B1 (ko) Mrgprx2를 이용한 즉시형 과민 반응 환자를 판별하는 방법
US20220326243A1 (en) Composition for cancer diagnosis

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16858379

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16858379

Country of ref document: EP

Kind code of ref document: A1