Method to predict response to pharmacological chaperone treatment of diseases

Abstract

The present invention provides methods to determine whether a patient with a lysosomal storage disorder will benefit from treatment with a specific pharmacological chaperone. The present invention exemplifies an in vitro method for determining α-galactosidase A responsiveness to a pharmacological chaperone such as 1-deoxygalactonojirimycin in a cell line expressing a mutant from of α-galactosidase A. The invention also provides a method for diagnosing Fabry disease in patients suspected of having Fabry disease.

1 . A method for determining whether a patient expressing a mutant form of α-galactosidase A will respond to treatment with a specific pharmacological chaperone for the α-galactosidase, which method comprises
 a. contacting a first host cell with a pharmacological chaperone specific for α-galactosidase A, wherein the first host cell expresses the mutant form of α-galactosidase A; and 
 b. comparing α-galactosidase A activity in a second host cell not contacted with the specific pharmacological chaperone, with α-galactosidase A activity in the first host cell contacted with the specific pharmacological chaperone, 
 
       wherein an increase in α-galactosidase A activity in the first host cell contacted with the specific pharmacological chaperone compared to the activity of α-galactosidase A expressed by the second host cell not contacted with the specific pharmacological chaperone indicates that the patient will respond to treatment with the specific pharmacological chaperone, and wherein the mutant form of α-galactosidase A is selected from the group consisting of the α-galactosidase A mutations A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, G85D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L16H, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M96I, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C. 
     
     
         2 . The method of  claim 1 , wherein the mutant form of α-galactosidase A is caused by a missense mutation in a gene encoding α-galactosidase A. 
     
     
         3 . The method of  claim 1 , wherein the patient has been diagnosed with a lysosomal storage disorder. 
     
     
         4 . The method of  claim 3 , wherein the lysosomal storage disorder is Fabry disease. 
     
     
         5 . The method of  claim 1 , wherein the specific pharmacological chaperone is 1-deoxygalactonorjirimycin. 
     
     
         6 . The method of  claim 1 , wherein the host cell is selected from the group consisting of CHO cells, HeLa cells, HEK-293 cells, GripTite 293 MSR cells, 293T cells, COS cells, COS-7 cells, mouse primary myoblasts, and NIH 3T3 cells. 
     
     
         7 . The method of  claim 6 , wherein the host cell is a HEK-293 cell. 
     
     
         8 . The method of  claim 6 , wherein the host cell is a COS-7 cell. 
     
     
         9 . The method of  claim 6 , wherein α-galactosidase A activity is determined using a fluorometric assay that quantifies hydrolysis of substrate in lysates from the host cell. 
     
     
         10 . A method of treating a patient diagnosed with Fabry disease which comprises administering to the patient a therapeutically effective dose of 1-deoxygalactonorjirimycin, wherein the patient expresses a mutant α-galactosidase A selected from the group consisting of the α-galactosidase A mutations A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, G85D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L16H, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M96I, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C. 
     
     
         11 . The method of  claim 10 , wherein the lysosomal storage disorder is Fabry disease. 
     
     
         12 . The method of  claim 10 , wherein the specific pharmacological chaperone is 1-deoxygalactonorjirimycin. 
     
     
         13 . A method of treating a patient diagnosed with Fabry disease, which method comprises administering to the patient a therapeutically effective dose of 1-deoxygalactonorjirimycin, wherein the patient expresses a mutation in α-galactosidase A, with the proviso that the mutation is not a nonresponsive mutation. 
     
     
         14 . A method of treating a patient diagnosed with Fabry disease, which method comprises administering to the patient a therapeutically effective dose of 1-deoxygalactonorjirimycin, wherein the patient expresses a mutation in α-galactosidase A, with the proviso that the mutation is not a mutation in which no enzyme is expressed. 
     
     
         15 . A kit for treating Fabry disease comprising at least one specific pharmacological chaperone for α-galactosidase and instructions to administer the specific pharmacological chaperone to a patient having at least one of a mutant form of α-galactosidase selected from the group consisting of the α-galactosidase A mutations A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, G85D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L161I, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M96I, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C. 
     
     
         16 . The kit of  claim 15 , wherein the instructions list at least 10% of the following α-galactosidase mutations selected from A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, G85D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L16H, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M961, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C. 
     
     
         17 . (canceled) 
     
     
         18 . The kit of  claim 15 , wherein the instructions list at least 25% of the following α-galactosidase mutations selected from A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, G85D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L16H, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M96I, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C. 
     
     
         19 . The kit of  claim 15 , wherein the instructions list at least 50% of the following α-galactosidase mutations selected from A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, G85D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L16H, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M961, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C. 
     
     
         20 . The kit of  claim 15 , wherein the instructions include at least 75% of mutant form of α-galactosidase selected from the group consisting of the α-galactosidase A mutations A257D, A257G, A257P, A291T, A292T, A307T, A309P, A352V, A368T, A73V, A97V, C174G, C174R, C56F, C56Y, D165H, D313G, D322E, D55V, E203V, F169S, G171R, G183A, G183V, G258R, G258V, G261D, G325S, G360D, G360S, 085D, G85M, I117S, I198T, I239T, I253T, I289S, I319T, I359T, K185E, K308N, L166G, L16H, L19P, L243W, L36F, L36S, L372P, L403S, L54P, M290I, M290L, M296L, M296T, M42L, M42R, M76T, M961, N53L, P205S, P293T, P409A, P409T, P60L, Q107L, Q250P, Q312R, Q321H, Q321L, R301G, R356G, S238N, S247C, T282A, T410I, V339E, W162G, W349S, Y152C, Y184C, Y200C, Y207H and Y216C.