Methods of Determining Efficacy of Therapy For Niemann-Pick C Disease And Related Disorders

Abstract

A method for identifying and screening subjects for cholesterol storage or trafficking diseases using an oxysterol as a biomarker. More particularly, subjects are screened and can be identified as having Niemann-Pick C disease, the method comprising the steps quantifying the concentration of an oxysterol in a biological sample taken from the subject and comparing the concentration of the oxysterol of the subject to a reference value of the oxysterol derived from a non-affected subject population. If the concentration of the oxysterol from the subject is higher than the reference value, the subject is identified as affected with Niemann-Pick C disease.

What is claimed is: 
     
         1 . A method for determining efficacy of a therapy in a subject afflicted with a disorder involving the accumulation of one or more oxysterols, comprising:
 (a) providing at least one first body fluid sample from the subject prior to administering a therapy to the subject;   (b) administering the therapy to the subject;   (c) obtaining at least one second body fluid sample from the subject after the administering the therapy;   (d) subjecting the at least one first body fluid sample to a first chromatography-mass spectroscopy analysis to determine concentration in the at least one first body fluid sample of at least one oxysterol selected from the group consisting of 7-ketocholesterol and cholestane-3β,5α,6β-triol;   (e) subjecting the at least one second body fluid sample to a second chromatography-mass spectroscopy analysis to determine concentration in the at least one second body fluid sample of the at least one oxysterol; and   (f) determining magnitude of difference between the at least one oxysterol concentration of the at least one first body fluid sample and the at least one second body fluid sample, whereby a decrease in concentration of the at least one oxysterol in the second sample compared to the first sample indicates efficacy of the therapy.   
     
     
         2 . A method in accordance with  claim 1 , wherein the therapy is selected from the group consisting of enzyme replacement therapy, gene therapy, statin administration, administration of a cholesterol synthesis inhibitor, administration of a liver X receptor (LXR) agonist, administration of an orphan nuclear receptor PXR ligands, administration of a neurosteroid, administration of N-butyldeoxynojirimycin (NB-DNJ) (miglustat), administration of curcumin, administration of an antibiotic, administration of salsalate, administration of salicylic acid, administration of an RXR ligand, administration of a sphingolipid synthesis inhibitor (myriocin), administration of KCl, administration of EGTA, administration of a calcium channel inhibitor, administration of nifedipine, administration of verapamil, administration of an antioxidant, administration of vitamin E, administration of vitamin C, administration of aurintricarboxylic acid, administration of flavonoids, administration of an estrogen, administration of propyl gallate, administration of glutathione, administration of a caspase inhibitor, administration of a MAP kinase inhibitor, administration of a peroxisome proliferator-activated receptor (PPARs) ligand, administration of 15d-PGJ2, administration of WY 14643, administration of indomethacin, administration of a gluccorticoid, administration of dexamethasone, administration of hydrocortisone, administration of a PI-3 kinase inhibitor, administration of an NMDA open channel blocker, dietary therapy and a combination thereof. 
     
     
         3 . A method in accordance with  claim 1 , wherein the at least one oxysterol is cholestane-3β,5α,6β-triol. 
     
     
         4 . A method in accordance with  claim 1 , wherein the at least one oxysterol is 7-ketocholesterol. 
     
     
         5 . A method in accordance with  claim 4 , wherein the therapy is cyclodextrin therapy. 
     
     
         6 . A method in accordance with  claim 1 , wherein each of the first chromatography-mass spectroscopy analysis and the second chromatography-mass spectroscopy analysis comprises:
 (a) adding to the body fluid sample a known amount of an internal standard comprising an oxysterol other than the at least one oxysterol;   (b) isolating the at least one oxysterol from the biological sample; and   (c) quantifying the isolated at least one oxysterol.   
     
     
         7 . A method in accordance with  claim 6 , wherein the quantifying the at least one oxysterol comprises determining the relative concentration of the at least one oxysterol and the known amount of internal standard in the body fluid sample by correlating the area under the curve obtained for the at least one oxysterol with the area under the curve obtained for the internal standard. 
     
     
         8 . A method in accordance with  claim 1 , wherein the first biological sample and the second biological sample are each selected from the group consisting of plasma, serum, blood, sputum and amniotic fluid. 
     
     
         9 . A method in accordance with  claim 1 , wherein the disorder involving the accumulation of one or more oxysterols is selected from the group consisting of a lysosomal storage disease, a cholesterol trafficking disease, a neurodegenerative disease and a combination thereof. 
     
     
         10 . A method in accordance with  claim 1 , wherein the disorder involving the accumulation of one or more oxysterols is selected from the group consisting of Niemann-Pick C (NPC) disease, infantile neuronal ceroid lipofuscinosis, GM1 gangliosidosis (“GM-1”), GM-2 gangliosidosis (“GM-2”), Gaucher's disease and hepatosplenomegaly. 
     
     
         11 . A method in accordance with  claim 1 , wherein the disorder involving the accumulation of an oxysterol is Niemann-Pick C (NPC) disease. 
     
     
         12 . A method for determining efficacy of a therapy in a subject afflicted with a disorder involving the accumulation of one or more oxysterols, comprising:
 (a) obtaining at least one first body fluid sample from the subject prior to administering a therapy to the subject;   (b) administering the therapy to the subject;   (c) obtaining at least one second body fluid sample from the subject after the administering the therapy;   (d) subjecting the at least one first body fluid sample to a first chromatography-mass spectroscopy analysis to determine concentration in the at least one first body fluid sample of 24-hydroxycholesterol;   (e) subjecting the at least one second body fluid sample to a second chromatography-mass spectroscopy analysis to determine concentration in the at least one second body fluid sample of 24-hydroxycholesterol; and   (f) determining magnitude of difference between the 24-hydroxycholesterol concentration of the at least one first body fluid sample and the at least one second body fluid sample, whereby an increase in concentration of 24-hydroxycholesterol in the second sample compared to the first sample indicates efficacy of the therapy.   
     
     
         13 . A method in accordance with  claim 12 , wherein the therapy is selected from the group consisting of enzyme replacement therapy, gene therapy, statin administration, administration of a cholesterol synthesis inhibitor, administration of a liver X receptor (LXR) agonist, administration of an orphan nuclear receptor PXR ligands, administration of a neurosteroid, administration of N-butyldeoxynojirimycin (NB-DNJ) (miglustat), administration of curcumin, administration of an antibiotic, administration of salsalate, administration of salicylic acid, administration of an RXR ligand, administration of a sphingolipid synthesis inhibitor (myriocin), administration of KCl, administration of EGTA, administration of a calcium channel inhibitor, administration of nifedipine, administration of verapamil, administration of an antioxidant, administration of vitamin E, administration of vitamin C, administration of aurintricarboxylic acid, administration of flavonoids, administration of an estrogen, administration of propyl gallate, administration of glutathione, administration of a caspase inhibitor, administration of a MAP kinase inhibitor, administration of a peroxisome proliferator-activated receptor (PPARs) ligand, administration of 15d-PGJ2, administration of WY 14643, administration of indomethacin, administration of a glucocorticoid, administration of dexamethasone, administration of hydrocortisone, administration of a PI-3 kinase inhibitor, administration of an NMDA open channel blocker, dietary therapy and a combination thereof. 
     
     
         14 . A method in accordance with  claim 12 , wherein each of the first chromatography-mass spectroscopy analysis and the second chromatography-mass spectroscopy analysis comprises:
 (a) adding to the body fluid sample a known amount of an internal standard comprising an oxysterol other than 24-hydroxycholesterol;   (b) isolating 24-hydroxycholesterol from the biological sample; and   (c) quantifying the isolated 24-hydroxycholesterol.   
     
     
         15 . A method in accordance with  claim 15 , wherein the quantifying the 24-hydroxycholesterol comprises determining the relative concentration of the 24-hydroxycholesterol and the known amount of internal standard in the body fluid sample by correlating the area under the curve obtained for the 24-hydroxycholesterol with the area under the curve obtained for the internal standard. 
     
     
         16 . A method in accordance with  claim 12 , wherein the first biological sample and the second biological sample are each selected from the group consisting of plasma, serum, blood, sputum and amniotic fluid. 
     
     
         17 . A method in accordance with  claim 12 , wherein the disorder involving the accumulation of one or more oxysterols is selected from the group consisting of a lysosomal storage disease, a cholesterol trafficking disease, a neurodegenerative disease and a combination thereof. 
     
     
         18 . A method in accordance with  claim 12 , wherein the disorder involving the accumulation of one or more oxysterols is selected from the group consisting of Niemann-Pick C (NPC) disease, infantile neuronal ceroid lipofuscinosis, GM1 gangliosidosis (“GM-1”), GM-2 gangliosidosis (“GM-2”), Gaucher's disease and hepatosplenomegaly. 
     
     
         19 . A method in accordance with  claim 12 , wherein the disorder involving the accumulation of an oxysterol is Niemann-Pick C (NPC) disease. 
     
     
         20 . A method in accordance with  claim 12 , wherein the disorder involving the accumulation of an oxysterol is Tay-Sachs disease.