Innate immune proteins as biomarkers for cns injury

Abstract

The present invention provides novel markers of the severity of a central nervous system injury, such as spinal cord injury or traumatic brain injury, in a patient. In particular, protein components of inflammasomes in the cerebrospinal fluid that can be used to assess the severity of central nervous system injury in a patient are disclosed. Methods of using such protein biomarkers to determine a prognosis, direct treatment and rehabilitation efforts, and monitor response to treatment for a patient with a central nervous system injury are also described.

1 . A method of evaluating a patient suspected of having a central nervous system (CNS) injury comprising:
 providing a biological sample from a patient presenting with clinical symptoms consistent with a CNS injury;   measuring the level of at least one inflammasome protein in the biological sample;   determining the presence or absence of a protein signature associated with a CNS injury or a more severe CNS injury, wherein the protein signature comprises an elevated level of said at least one inflammasome protein; and   selecting patients exhibiting the presence of the protein signature as having a CNS injury or a more severe CNS injury.   
     
     
         2 . The method of  claim 1 , wherein the level of said at least one inflammasome protein in the protein signature is enhanced relative to the level of said at least one inflammasome protein in a control sample. 
     
     
         3 . The method of  claim 1 , wherein the level of said at least one inflammasome protein in the protein signature is enhanced relative to a pre-determined reference value or range of reference values. 
     
     
         4 . The method of  claim 1 , wherein the CNS injury is a spinal cord injury or traumatic brain injury. 
     
     
         5 . The method of  claim 1 , wherein said at least one inflammasome protein is nucleotide-binding leucine-rich repeat pyrin domain containing protein 1 (NLRP1), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase-1, or combinations thereof. 
     
     
         6 . The method of  claim 6 , wherein the protein signature comprises an elevated level for each of caspase-1, NLRP1, and ASC. 
     
     
         7 . The method of  claim 6 , wherein the protein signature comprises an elevated level for the p20 subunit of caspase-1. 
     
     
         8 . The method of  claim 1 , wherein said biological sample is obtained within one week of the suspected injury. 
     
     
         9 . The method of  claim 1 , wherein said biological sample is obtained within five days of the suspected injury. 
     
     
         10 . The method of  claim 1 , wherein said biological sample is obtained within three days of the suspected injury. 
     
     
         11 . The method of  claim 1 , wherein said biological sample is cerebrospinal fluid (CSF), CNS microdialysate, saliva, serum, plasma, or urine. 
     
     
         12 . The method of  claim 1 , wherein the patient has an A or B rating on the American Spinal Cord Injury Association (ASIA) Impairment Scale. 
     
     
         13 . The method of  claim 1 , wherein the patient has a Glasgow Coma Scale (GCS) score of 3 to 12. 
     
     
         14 . The method of  claim 13 , wherein the patient has a GCS score of 3 to 8. 
     
     
         15 . The method of  claim 1 , wherein the patient is a pediatric patient. 
     
     
         16 . The method of  claim 1 , wherein the level of said at least one inflammasome protein is measured by immunoblot or ELISA. 
     
     
         17 . The method of  claim 1  further comprising administering a neuroprotective treatment to the patient when said protein signature is identified. 
     
     
         18 . The method of  claim 17 , where in the neuroprotective treatment is hypothermia, methylprednisolone, 17α-estradiol, 17β-estradiol, ginsenoside, progesterone, simvastatin, deprenyl, minocycline, and resveratrol. 
     
     
         19 . The method of  claim 17 , further comprising:
 measuring the level of said at least one inflammasome protein in a biological sample obtained from the patient following neuroprotective treatment;   preparing a treatment protein signature associated with a positive response to the neuroprotective treatment, wherein the treatment protein signature comprises a reduced level of at least one inflammasome protein; and   identifying patients exhibiting the presence of the treatment protein signature as responding positively to the neuroprotective treatment.   
     
     
         20 . A kit for preparing an inflammasome protein profile associated with CNS injury comprising a labeled-binding partner that specifically binds to one or more inflammasome proteins, wherein said one or more inflammasome proteins are selected from the group consisting of NLRP1, ASC, caspase-1, and combinations thereof. 
     
     
         21 . The kit of  claim 20 , wherein the kit comprises labeled-binding partners that specifically binding to each of NLRP1, ASC, and caspase-1. 
     
     
         22 . The kit of  claim 20 , wherein the labeled-binding partner is a labeled-antibody or fragment thereof, aptamer, or peptide. 
     
     
         23 . The kit of  claim 20 , wherein the binding partner is labeled with metal nanoparticles, a fluorescent label, or an enzyme label. 
     
     
         24 . A method of determining a prognosis for a patient with a CNS injury comprising:
 providing a biological sample obtained from the patient within a week of injury; and   measuring the level of at least one inflammasome protein in the biological sample to prepare an inflammasome protein profile, wherein the inflammasome protein profile is indicative of the prognosis of the patient.   
     
     
         25 . The method of  claim 24 , wherein an elevated level of at least one inflammasome protein relative to a pre-determined reference value or range of reference values is predictive of the patient having a Glasgow Outcome Scale (GOS) score of 1 to 3 upon follow-up assessment. 
     
     
         26 . The method of  claim 24 , wherein said at least one inflammasome protein is NLRP1, ASC, caspase-1, or combinations thereof. 
     
     
         27 . The method of  claim 26 , wherein said at least one inflammasome protein is the p20 subunit of caspase-1. 
     
     
         28 . The method of  claim 24 , wherein the CNS injury is a spinal cord injury or traumatic brain injury. 
     
     
         29 . The method of  claim 24 , wherein a reduced level of at least one inflammasome protein relative to a pre-determined reference value or range of reference values is predictive of the patient having a GOS score of 4 or 5 upon follow-up assessment. 
     
     
         30 . The method of  claim 24 , wherein said biological sample is CSF, CNS microdialysate, saliva, serum, plasma, or urine. 
     
     
         31 . The method of  claim 24 , wherein the level of said at least one inflammasome protein is measured by immunoblot or ELISA. 
     
     
         32 . The method of  claim 24 , wherein the patient is a pediatric patient.