Differential expression of molecules associated with acute stroke

Abstract

Methods are provided for evaluating a stroke, for example for determining whether a subject has had an ischemic stroke, determining the severity or likely neurological recovery of a subject who has had an ischemic stroke, and determining a treatment regimen for a subject who has had an ischemic stroke, as are arrays and kits that can be used to practice the methods. In particular examples, the method includes screening for expression in ischemic stroke related genes (or proteins), such as white blood cell activation and differentiation genes (or proteins), genes (or proteins) related to hypoxia, genes (or proteins) involved in vascular repair, and genes (or proteins) related to a specific peripheral blood mononuclear cell (PBMC) response to the altered cerebral microenvironment. Also provided are methods of identifying one or more agents that alter the activity (such as the expression) of an ischemic stroke-related molecule.

1 . A method of determining whether a human subject has had an ischemic stroke, comprising:
 detecting expression of ischemic stroke-related nucleic acid molecules of the subject obtained from a blood sample, wherein the ischemic stroke-related molecules comprise at least four ischemic stroke-related molecules represented by any combination of at least four molecules listed in any of Tables 2-5;   comparing expression of the ischemic stroke-related nucleic acid molecules to a control representing expression of the ischemic stroke-related nucleic acid molecules expected in a subject who has not had an ischemic stroke; and   determining that the subject has had an ischemic stroke when the presence of an at least 4-fold change in expression of the ischemic stroke-related nucleic acid molecules in the subject relative to the control is detected.   
     
     
         2 . The method of  claim 1 , wherein expression of ischemic stroke-related nucleic acid molecules is detected within 24 hours of onset of clinical signs and symptoms that indicate a potential stroke. 
     
     
         3 . The method of  claim 1 , wherein expression of ischemic stroke-related nucleic acid molecules is detected within 7-14 days of onset of clinical signs and symptoms that indicate a potential stroke. 
     
     
         4 . The method of  claim 1 , wherein expression of ischemic stroke-related nucleic acid molecules is detected within 90 days of onset of clinical signs and symptoms that indicate a potential stroke. 
     
     
         5 . The method of  claim 1 , wherein the at least four ischemic-stroke related molecules comprise CD163, hypothetical protein FLJ22662 Laminin A motif; adrenomedullin; KIAA0146 protein; amyloid β(A4) precursor-like protein; CD36; baculoviral IAP repeat-containing protein 1; or combinations thereof. 
     
     
         6 . The method of  claim 1 , wherein the method comprises determining whether there is increased expression any combination of at least four ischemic stroke-related genes, wherein the presence of increased expression of the at least four ischemic stroke-related molecules indicates that the subject has had an ischemic stroke. 
     
     
         7 . The method of  claim 1 , wherein the method has a sensitivity of at least 78% and accuracy of at least 80%. 
     
     
         8 . The method of  claim 1 , wherein the subject had an onset of clinical signs and symptoms of an ischemic stroke no more than 72 hours prior to detecting expression of the at least four ischemic stroke-related molecules. 
     
     
         9 . The method of  claim 1 , where the nucleic acid molecules comprise mRNA or cDNA. 
     
     
         10 . The method of  claim 1 , wherein the nucleic acid molecules are isolated from the subject, thereby generating isolated nucleic acid molecules, and wherein the isolated nucleic acid molecules are hybridized with oligonucleotides that detect the at least four ischemic stroke-related molecules. 
     
     
         11 . The method of  claim 10 , wherein the oligonucleotides are present on an array substrate. 
     
     
         12 . The method of  claim 1 , wherein the nucleic acid molecules isolated from the blood sample are obtained from peripheral blood mononuclear cells (PBMCs). 
     
     
         13 . The method of  claim 1 , wherein the isolated nucleic acid molecules are labeled with a detectable label. 
     
     
         14 . The method of  claim 11 , wherein the oligonucleotides are labeled with a detectable label. 
     
     
         15 . The method of  claim 1 , wherein the blood sample is obtained from the subject within 24 hours of onset of clinical indications of stroke. 
     
     
         16 . The method of  claim 1 , further comprising administering to the subject a treatment to avoid or reduce ischemic injury if the expression indicates that the subject has had an ischemic stroke. 
     
     
         17 . The method of  claim 16 , wherein the selected treatment comprises treating the subject with an anticoagulant agent, a thrombolytic agent, or combinations thereof. 
     
     
         18 . The method of  claim 1 , wherein detecting expression comprises quantifying expression of the at least four ischemic stroke-related molecules. 
     
     
         19 . An array comprising oligonucleotides complementary to ischemic stroke-related gene sequences, wherein the ischemic stroke-related gene sequences comprise any combination of at least four of the genes listed in Tables 2-5. 
     
     
         20 . A method of determining whether a human subject has had an ischemic stroke, comprising:
 applying isolated nucleic acid molecules obtained from PBMCs of the subject to an array, wherein the array consists of oligonucleotides complementary to at least four ischemic stroke-related molecules represented by any combination of at least four molecules listed in any of Tables 2-5;   incubating the isolated nucleic acid molecules with the array for a time sufficient to allow hybridization between the isolated nucleic acid molecules and oligonucleotide probes, thereby forming isolated nucleic acid molecule:oligonucleotide complexes; and   analyzing the isolated nucleic acid molecule:oligonucleotide complexes to determine if expression of the isolated nucleic acid molecules is altered, wherein the presence of differential expression in the at least four ischemic stroke-related molecules indicates that the subject has had an ischemic stroke.