Biomarkers for h-nox delivery of oxygen

Abstract

The invention provides methods to monitor tumor oxygenation by H-NOX proteins. H-NOX proteins extravasate into and preferentially accumulate in tumor tissue for sustained delivery of oxygen. For example, the invention provides methods to monitor brain tumor oxygenation by H-NOX proteins for enhanced treatment of brain cancers.

What is claimed is: 
     
         1 . A method of treating a hypoxic brain tumor in an individual comprising
 a) administering an effective amount of an H-NOX protein to the individual,   b) determining the level of hypoxia in the brain tumor following administration of the H-NOX protein, and   c) administering an effective amount of radiation to the individual wherein the tumor hypoxia measured in step b) is reduced compared to the level of hypoxia in the brain tumor prior to H-NOX administration.   
     
     
         2 . A method of treating a hypoxic brain tumor in an individual comprising
 a) determining the level of hypoxia in the brain tumor,   b) administering an effective amount of an H-NOX protein to the individual,   c) determining the level of hypoxia in the brain tumor following administration of the H-NOX protein, and   d) administering an effective amount of radiation to the individual wherein the tumor hypoxia measured in step c) is reduced compared to the level of hypoxia measured in step a).   
     
     
         3 . A method of optimizing therapeutic efficacy for treatment of a hypoxic brain tumor in an individual, the method comprising
 a) administering H-NOX to the individual,   b) measuring the level of hypoxia of the tumor one or more times after administration of the H-NOX protein,   c) administering radiation therapy when tumor hypoxia is reduced compared to the level of hypoxia prior to H-NOX administration.   
     
     
         4 . The method of  claim 3 , wherein the hypoxia is reduced by at least about 5%, 10%, 15%, 20%, 25% or 50%. 
     
     
         5 . A method of monitoring the efficacy of delivery of O 2  to hypoxic brain tumor by an H-NOX protein in an individual, the method comprising
 a) administering an effective amount of H-NOX protein to the individual,   b) measuring the level of hypoxia in the tumor at one or more time points after administration of the H-NOX protein, wherein a reduction of tumor hypoxia compared to the level of hypoxia in the tumor prior to administration of H-NOX indicates effective delivery of O 2  to the brain tumor.   
     
     
         6 . The method of  claim 5 , wherein the reduction in tumor hypoxia indicates that the individual is suitable for administration of radiation therapy. 
     
     
         7 . The method of  claim 3 , wherein the level of hypoxia in the tumor is measured one or more of one hour, two hours, three hours, four hours, eight hours, twelve hours, 24 hours, 48 hours or 72 hours after administration of H-NOX. 
     
     
         8 . A method of monitoring responsiveness or lack of responsiveness to treatment with a H-NOX in an individual suffering from a brain tumor comprising measuring the hypoxic state of the tumor following H-NOX administration, wherein responsiveness is indicated by a reduction in tumor hypoxia. 
     
     
         9 . The method of  claim 8 , wherein the responsiveness indicates that the individual is suitable for administration of radiation therapy. 
     
     
         10 . A method of identifying an individual with a brain tumor who is more likely to exhibit benefit from a therapy comprising an H-NOX protein, said method comprising
 a) determining the hypoxia level of the tumor,   a) administering H-NOX to the individual,   b) measuring the level of hypoxia of the tumor,   c) wherein about a 5% decrease in hypoxia indicates the individual is more likely to exhibit benefit from radiation treatment in combination with H-NOX treatment.   
     
     
         11 . The method of  claim 10 , wherein the decrease in hypoxia of step c) is a at least a 10%, a 15%, a 20%, a 25%, a 50%, a 75% or a 100% decrease in hypoxia. 
     
     
         12 . The method of  claim 1 , wherein tumor hypoxia is measured by one or more of  18 F-fluoromisonidazole (FMISO) tumor uptake, pimidazole uptake,  18 F-fluoroazomycin arabinoside (FAZA) uptake, a nitroimidazole uptake, Copper(II)-diacetyl-bis(N 4 -methylthiosemicarbazone (Cu-ATSM) uptake,  19 F magnetic resonance imaging of hexafluorobenzene (C6F6) uptake,  1 H MRI of hexamethyldisiloxane uptake, tumor HIF-1α expression, tumor Glut-1 expression, tumor LDHA expression, tumor carbonic anhydrase IX (CA-9) expression, or lactate and/or pyruvate levels. 
     
     
         13 . The method of  claim 1 , wherein the determination of the level of hypoxia in the tumor is repeated. 
     
     
         14 - 15 . (canceled) 
     
     
         16 . The method of claim  15 , further comprising administration of radiation following administration of H-NOX. 
     
     
         17 . (canceled) 
     
     
         18 . The method of  claim 1 , wherein the radiation is X-radiation. 
     
     
         19 . The method of  claims 18 , wherein the X-radiation is administered at about 0.5 gray to about 75 gray. 
     
     
         20 . The method of  claim 1 , where the brain cancer is glioblastoma. 
     
     
         21 . (canceled) 
     
     
         22 . The method of  claim 1 , wherein the individual is a human. 
     
     
         23 - 24 . (canceled) 
     
     
         25 . The method of  claim 1 , wherein the H-NOX protein is a  T. tengcongensis  H-NOX, a  L. pneumophilia  2 H-NOX, a  H. sapiens β 1, a  R. norvegicus β 1, a  C. lupus  H-NOX domain, a  D. melangaster  β1, a  D. melangaster  CG14885-PA, a  C. elegans  GCY-35, a  N. punctiforme  H-NOX,  C. crescentus  H-NOX, a  S. oneidensis  H-NOX, or  C. acetobutylicum  H-NOX. 
     
     
         26 . The method of  claim 1 , wherein the H-NOX protein comprises a H-NOX domain corresponding to the H-NOX domain of  T. tengcongensis  set forth in SEQ ID NO:2. 
     
     
         27 . The method of  claim 1 , wherein the H-NOX comprises one or more distal pocket mutations. 
     
     
         28 . The method of  claim 27 , wherein the distal pocket mutation is an amino acid substitution at a site corresponding to L144 of  T. tengcongensis  H-NOX. 
     
     
         29 . (canceled) 
     
     
         30 . The method of  claim 26 , wherein the amino acid substitution at position 144 is an L144F substitution. 
     
     
         31 - 34 . (canceled) 
     
     
         35 . The method of  claim 1 , wherein the H-NOX protein is a polymeric H-NOX protein. 
     
     
         36 . The method of  claim 35 , wherein the polymeric H-NOX protein comprises monomers, wherein the monomers comprise an H-NOX domain and a polymerization domain. 
     
     
         37 . The method of  claim 36 , wherein the H-NOX domain is covalently linked to the polymerization domain. 
     
     
         38 . The method of  claim 35 , wherein the polymeric H-NOX protein is a trimeric H-NOX protein. 
     
     
         39 . The method of  claim 38 , wherein the trimeric H-NOX protein comprises one or more trimerization domains. 
     
     
         40 . (canceled) 
     
     
         41 . The method of  claim 39 , wherein the trimerization domain is a foldon domain. 
     
     
         42 . The method of  claim 41 , wherein the foldon domain comprises the amino acid sequence of SEQ ID NO:4. 
     
     
         43 . (canceled) 
     
     
         44 . The method of  claim 1 , wherein the H-NOX protein is covalently bound to polyethylene glycol. 
     
     
         45 - 56 . (canceled)