Peripheral marker of blood brain barrier permeability

• US 20030170747A1
• Filed: 06/25/2001
• Published: 09/11/2003
• Est. Priority Date: 06/25/2001
• Status: Active Grant

• Current Assignee: Cleveland Clinic Foundation
• Original Assignee: Cleveland Clinic Foundation
• Inventors: Janigro, Damir, Barnett, Gene, Mayberg, Marc
• Granted Patent: US 6,884,591 B2
• Time in Patent Office: Days
• US Class Current: 435/7.21
• CPC Class Codes: G01N 33/574 for cancer

Abstract

The present invention relates generally to a peripheral marker or markers of blood brain barrier (“BBB”) integrity and methods of using same in the diagnosis, prognosis, and treatment of a variety of diseases. The peripheral marker(s) of the present invention are particularly useful in the differential diagnosis of diseased states. The preferred embodiments of the present invention relate to methods, compositions, kits, and assays useful in determining the integrity or permeability of a blood brain barrier. The various embodiments of the present invention can be used to identify subjects at risk for developing a disease associated with increased permeability of the blood brain barrier, as well as to provide insight on the ability of an agent or agents to pass the blood brain barrier. Embodiments of the present invention preferably involve the use of subject derived blood samples to determine the occurrence and level of circulating proteins indicative of blood brain barrier permeability or integrity. The embodiments of the present invention also provides screening methods for diagnosis, prognosis, susceptibility, or degree of permeability of penetration of the blood brain barrier by detecting the presence of serum S-100β either directly or through the use of antibodies. The present invention further provides for kits for carrying out the above described screening methods. Preferably, such kits will be used to screen patients for increased levels of S100β protein alone or in combination with other markers of diseased states as a diagnostic and prognostic indicator of permeability of the BBB. Thus, the present invention provides a minimally invasive alternative to direct cerebrospinal fluid sampling to determine the permeability of the blood brain barrier.

Claims

1. A method for diagnosis of blood brain barrier permeability in a subject comprising:

detecting levels of S100β

protein in a blood sample derived from a subject; and

comparing the level of S100β

protein detected in the sample to a level of S100β

protein in a control, wherein an increase in the level of S100β

protein detected in the sample as compared to the control sample is indicative of blood brain barrier permeability.

2. The method of claim 1, wherein the sample is scored based upon permeability of the blood brain barrier.

3. The method of claim 1, wherein the S100β

protein is detected using an immunoassay.

4. The method of claim 3, wherein the immunoassay is an immunoprecipitation assay.

5. The method of claim 1, wherein the levels of S100β

protein in blood samples over time indicates stages of diseased states.

6. The method of claim 1, wherein the indication of blood brain permeability is made independent of indicators of neuronal distress.

7. The method of claim 1, further including detecting levels of markers of neuronal distress, said markers being selected from the group consisting of NSE, GFAP, and elevated levels of S100β

protein beyond increased levels of S100β

.

8. A method of treating a patient in need thereof with a therapeutic agent, said method comprising:

administering an agent which causes blood brain barrier opening;

detecting elevated levels of S100β

protein in the patient'"'"'s blood to ensure blood brain barrier opening; and

administering the therapeutic agent.

10. The method of claim 8, wherein the step of detecting elevated levels of S100β

to ensure blood brain barrier opening is repeated over a course of treatment of a patient benefiting from said blood brain barrier opening.

11. The method of claim 8, wherein said therapeutic agent is selected from the group consisting of:

chemotherapeutics, pharmaceuticals, neuropharmaceuticals, potential neuropharmaceuticals, and other neurologically active agents.

9. The method of claim 9 further comprising the step of detecting reduced levels of S100β

protein in the patient'"'"'s blood to confirm a closed blood brain barrier.

12. A method for delivering a compound from the bloodstream to the brain, the method comprising:

introducing an agent into the bloodstream to open the blood brain barrier;

determining the level of S100β

in the blood; and

introducing the compound into the bloodstream when S100β

protein in the bloodstream is elevated.

13. The method according to claim 12, comprising admitting the compound into the patient'"'"'s bloodstream in a vicinity of the brain.

14. The method according to claim 12, wherein said compound is selected from the group consisting of a contrast agent, a neuropharmacologic agent, a neuroactive peptides, a protein, an enzyme, a gene therapy agent, a neuroprotective factor, a growth factor, a biogenic amine, a trophic factor to any of brain and spinal transplants, an immunoreactive proteins, a receptor binding protein, a radioactive agent, an antibody, and a cytotoxin.

15. The method of claim 12, wherein the step of determining the level of S100β

is repeated over a course of treatment to determine efficacy of the treatment.

16. The method of claim 12, further including the step of comparing to specific threshold values of the concentration of a S100β

protein to determine the presence of statistically significant concentrations thereof above normal levels.

17. A method of diagnosing cancer, said method comprising detecting a cancer marker if present;

and determining levels of S100β

in a patient'"'"'s blood, wherein elevated levels of S100β

in the patient'"'"'s blood is indicative of brain cancer.

18. The method of claim 17, wherein presence of elevated S100β

protein without detection of the cancer marker is indicative of a primary tumor.

19. The method of claim 17 wherein presence of elevated S100β

protein and a cancer marker from another organ is indicative of metastatic cancer.

20. The method of claim 17, wherein a patient predisposed to re-occurrence of a primary tumor is screened for elevated S100β

protein in the patient'"'"'s blood to indicate re-occurrence of the primary tumor.