Diagnostic markers of stroke and cerebral injury and methods of use thereof
The present invention relates to methods for the diagnosis and evaluation of stroke and transient ischemic attacks. In a particular aspect, patient samples are analyzed for the presence or amount of a panel of markers comprising one or more specific markers for cerebral injury and one or more non-specific markers for cerebral injury. In an alternative aspect, samples are analyzed for B-type natriuretic peptide. A variety of markers are disclosed for assembling a panel for such diagnosis and evaluation. In various aspects, the invention provides methods for early detection and differentiation of stroke types and transient ischemic attacks, for determining the prognosis of a patient presenting with stroke symptoms, and identifying a patient at risk for cerebral vasospasm. Invention methods provide rapid, sensitive and specific assays to greatly increase the number of patients that can receive beneficial stroke treatment and therapy, and reduce the costs associated with incorrect stroke diagnosis.
1. A method of determining a diagnosis of stroke or cerebral injury in a subject, said method comprising:
analyzing a test sample obtained from a subject for the presence or amount of one or more markers selected from the group consisting of adenylate kinase, brain-derived neurotrophic factor, calbindin-D, creatine kinase-BB, glial fibrillary acidic protein, lactate dehydrogenase, myelin basic protein, neural cell adhesion molecule, neuron-specific enolase, neurotrophin-3, one or more isoforms of protein kinase C, proteolipid protein, S-100β, thrombomodulin, and marker(s) related thereto, wherein said test sample is blood, serum, or plasma;
analyzing said test sample for the presence or amount of caspase-3 or marker(s) related thereto; and
correlating the presence or amount of the analyzed markers to the occurrence or nonoccurrence of a stroke or cerebral injury in said subject.
2. A method according to claim 1 , wherein said analyzed markers further comprise one or more acute phase reactant(s) selected from the group consisting of C-reactive protein, E-selectin, insulin-like growth factor-1, intercellular adhesion molecule-1, interleukin-1β, interleukin-1 receptor antagonist, interleukin-6, matrix metalloproteinase-3, matrix metalloproteinase-9, monocyte chemotactic protein-1, transforming growth factor β, tumor necrosis factor α, vascular cell adhesion molecule, and marker(s) related thereto.
3. A method according to claim 1 , wherein said analyzed markers further comprise matrix metalloproteinase-9 or marker(s) related thereto.
4. A method according to claim 1 , wherein said analyzed markers further comprise C-reactive protein or marker(s) related thereto.
5. A method according to claim 1 , wherein said analyzed markers comprise S-100 β or marker(s) related thereto.
6. A method according to claim 1 , wherein said analyzed markers further comprise D-dimer or marker(s) related thereto.
7. A method according to claim 1 , wherein said analyzed markers further comprise B-type natriuretic peptide or marker(s) related thereto.
8. A method according to claim 1 , wherein said analyzed markers comprise brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, neural cell adhesion molecule, vascular endothelial growth factor and the A1 domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
9. A method according to claim 1 , wherein said analyzed markers comprise brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, neural cell adhesion molecule, S-100β and the A1 domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
10. A method according to claim 1 , wherein said analyzed markers comprise brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, monocyte chemotactic protein-1, neural cell adhesion molecule, and the A1 domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
11. A method according to claim 1 , wherein said analyzed markers comprise brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, neural cell adhesion molecule, vascular endothelial growth factor, and the integrin domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
12. A method according to claim 1 , wherein said analyzed markers comprise brain-derived neurotrophic factor, caspase-3, glial fibrillary acidic protein, matrix metalloproteinase-9, S-100β, and the A1 domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
13. A method according to claim 1 , wherein said analyzed markers comprise brain-derived neurotrophic factor, caspase-3, glial fibrillary acidic protein, matrix metalloproteinase-9, neural cell adhesion molecule, and S-100β, optionally substituting one or more of said markers with marker(s) related thereto.
14. A method according to claim 1 , wherein said analyzed markers comprise B-type natriuretic peptide, caspase-3, matrix metalloproteinase-9, monocyte chemotactic protein- 1, neural cell adhesion molecule, S-100β, and the integrin domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
15. A method according to claim 1 , wherein said analyzed markers comprise B-type natriuretic peptide, caspase-3, glial fibrillary acidic protein, matrix metalloproteinase-9, monocyte chemotactic protein-1, neural cell adhesion molecule, S-100β,and the A1 domain of von Willebrand factor, optionally substituting one or more of said markers with marker(s) related thereto.
16. A method according to claim 1 , further comprising comparing the amount of said analyzed markers to the amount of said analyzed markers in normal individuals, wherein changes in said amounts in said test sample as compared to normal individuals is indicative of the occurrence of a stroke or cerebral injury in said subject.
17. A method according to claim 1 , wherein the method rules out the occurrence of a stroke or cerebral injury in said subject.
18. A method of determining a diagnosis of stroke or cerebral injury in a subject, said method comprising:
analyzing a test sample obtained from a subject by performing an assay that detects one or more markers selected from the group consisting of adenylate kinase, brain-derived neurotrophic factor, calbindin-D, creatine kinase-BB, glial fibrillary acidic protein, lactate dehydrogenase, myelin basic protein, neural cell adhesion molecule, neuron-specific enolase, neurotrophin-3, one or more isoforms of protein kinase C, proteolipid protein, S-100β, and thrombomodulin, wherein said test sample is blood, serum, or plasma;
analyzing said test sample by performing an assay that detects caspase-3; and
correlating the results of said assays to the occurrence or nonoccurrence of a stroke or cerebral injury in said subject.
19. A method according to claim 18 , wherein said method further comprises performing one or more assays that detect one or more acute phase reactant(s) selected from the group consisting of C-reactive protein, E-selectin, insulin-like growth factor-1, intercellular adhesion molecule-1, interleukin-1β, interleukin-1 receptor antagonist, interleukin-6, matrix metalloproteinase-3, matrix metalloproteinase-9, monocyte chemotactic protein-1, transforming growth factor β, tumor necrosis factor α, vascular cell adhesion molecule, and marker(s) related thereto.
20. A method according to claim 18 , wherein said method further comprises performing an assay that detects metalloproteinase-9.
21. A method according to claim 18 , wherein said method further comprises performing an assay that detects C-reactive protein.
22. A method according to claim 18 , wherein said method comprises performing an assay that detects S-100 β.
23. A method according to claim 18 , wherein said method further comprises performing an assay that detects D-dimer.
24. A method according to claim 18 , wherein said method further comprises performing an assay that detects B-type natriuretic peptide, NT-proBNP, or proBNP.
25. A method according to claim 18 , wherein said method comprises performing assays that detect brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, neural cell adhesion molecule, vascular endothelial growth factor and the A1 domain of von Willebrand factor.
26. A method according to claim 18 , wherein said method comprises performing assays that detect brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, neural cell adhesion molecule, S-100β and the A1 domain of von Willebrand factor.
27. A method according to claim 18 , wherein said method comprises performing assays that detect brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase- 9, monocyte chemotactic protein-1, neural cell adhesion molecule, and the A1domain of von Willebrand factor.
28. A method according to claim 18 , wherein said method comprises performing assays that detect brain-derived neurotrophic factor, caspase-3, matrix metalloproteinase-9, neural cell adhesion molecule, vascular endothelial growth factor, and the integrin domain of von Willebrand factor.
29. A method according to claim 18 , wherein said method comprises performing assays that detect brain-derived neurotrophic factor, caspase-3, glial fibrillary acidic protein, matrix metalloproteinase-9, S-100β, and the A1 domain of von Willebrand factor.
30. A method according to claim 18 , wherein said method comprises performing assays that detect brain-derived neurotrophic factor, caspase-3, glial fibrillary acidic protein, matrix metalloproteinase-9, neural cell adhesion molecule, and S-100β.
31. A method according to claim 18 , wherein said method comprises performing assays that detect B-type natriuretic peptide, caspase-3, matrix metalloproteinase-9, monocyte chemotactic protein-1, neural cell adhesion molecule, S-100β, and the integrin domain of von Willebrand factor.
32. A method according to claim 18 , wherein said method comprises performing assays that detect B-type natriuretic peptide, caspase-3, glial fibrillary acidic protein, matrix metalloproteinase-9, monocyte chemotactic protein-1 neural cell adhesion molecule, S-100β, and the A1 domain of von Willebrand factor.
33. A method according to claim 18 , further comprising comparing the amount of said analyzed markers to the amount of said analyzed markers in normal individuals, wherein changes in said amounts in said test sample as compared to normal individuals is indicative of the occurrence of a stroke or cerebral injury in said subject.
34. A method according to claim 18 , wherein the method rules out the occurrence of a stroke or cerebral injury in said subject.