Raav-guanylate cyclase compositions and methods for treating leber's congenital amaurosis-1 (lca1)

Abstract

Disclosed are viral vector compositions comprising polynucleotide sequences that express one or more biologically-active mammalian guanylate cyclase proteins. Also disclosed are methods for their use in preventing, treating, and/or ameliorating at least one or more symptoms of a disease, disorder, abnormal condition, or dysfunction resulting at least in part from a guanylate cyclase deficiency in vivo. In particular embodiments, the use of recombinant adeno-associated viral (rAAV) vectors to treat or ameliorate symptoms of Leber's congenital amaurosis, as well as other conditions caused by an absence or reduction in the expression of a functional retinal-specific guanylate cyclase 1 (retGC1).

1 . A recombinant adeno-associated viral (rAAV) vector comprising at least a first polynucleotide that comprises a photoreceptor-specific human rhodopsin kinase promoter, or a ubiquitous truncated chimeric CMV-chicken β-actin promoter, operably linked to at least a first nucleic acid segment that encodes at least a first mammalian guanylate cyclase protein, peptide, or polypeptide. 
     
     
         2 . The rAAV vector according to  claim 1 , wherein the at least a first nucleic acid segment encodes at least a first mammalian guanylate cyclase protein, peptide, or polypeptide that comprises at least a first contiguous amino acid sequence region that is at least about 90% identical to at least a first sequence region of at least 80 contiguous amino acids of a sequence as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         3 . The rAAV vector according to  claim 1 , wherein the at least a first nucleic acid segment encodes at least a first mammalian guanylate cyclase protein, peptide, or polypeptide that comprises at least a first contiguous amino acid sequence region that is at least about 92% identical to at least a first sequence region of at least 100 contiguous amino acids of a sequence as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         4 . The rAAV vector according to  claim 1 , wherein the at least a first mammalian guanylate cyclase protein, peptide, or polypeptide comprises at least a first contiguous amino acid sequence region that is at least about 95% identical to at least a first sequence region of at least 120 contiguous amino acids of a sequence as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         5 . The rAAV vector according to  claim 1 , wherein the at least a first mammalian guanylate cyclase protein, peptide, or polypeptide comprises at least a first contiguous amino acid sequence region that is at least about 98% identical to at least a first sequence region of at least 140 contiguous amino acids of a sequence as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         6 . The rAAV vector according to  claim 1 , wherein the at least a first mammalian guanylate cyclase protein, peptide, or polypeptide comprises at least an amino acid sequence that is at least about 98% identical to the amino acid sequence of any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         7 . The rAAV vector according to  claim 1 , wherein the at least a first mammalian guanylate cyclase protein, peptide, or polypeptide comprises the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         8 . (canceled) 
     
     
         9 . The rAAV vector according to  claim 1  wherein the at least a first mammalian guanylate cyclase protein, peptide, or polypeptide is biologically-active and comprises an at least 50 amino acid contiguous sequence from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11. 
     
     
         10 .- 14 . (canceled) 
     
     
         15 . The rAAV vector according to  claim 1 , wherein the rAAV vector is comprised within an adeno-associated viral particle and wherein the adeno-associated viral particle is a recombinant adeno-associated virus serotype 1 (rAAV1) particle, a recombinant adeno-associated virus serotype 2 (rAAV2) particle, recombinant adeno-associated virus serotype 3 (rAAV3) particle, a recombinant adeno-associated virus serotype 4 (rAAV4) particle, a recombinant adeno-associated virus serotype 5 (rAAV5) particle, a recombinant adeno-associated virus serotype 6 (rAAV6) particle, a recombinant adeno-associated virus serotype 7 (rAAV7) particle, a recombinant adeno-associated virus serotype 8 (rAAV8) particle, or a recombinant adeno-associated virus serotype 9 (rAAV) particle. 
     
     
         16 . The rAAV vector according to  claim 1 , wherein the vector is a self-complementary rAAV (scAAV). 
     
     
         17 . The rAAV vector according to  claim 1 , wherein the photoreceptor-specific human rhodopsin kinase promoter comprises a nucleic acid sequence that consists essentially of an at least 30 contiguous base pair sequence from SEQ ID NO:12, or the truncated chimeric CMV-chicken β-actin promoter comprises a nucleic acid sequence that consists essentially of an at least 70 contiguous base pair sequence from SEQ ID NO:13. 
     
     
         18 .- 19 . (canceled) 
     
     
         20 . The rAAV vector according to  claim 1 , wherein the photoreceptor-specific human rhodopsin kinase promoter comprises the nucleic acid sequence of SEQ ID NO:12, or the truncated chimeric CMV-chicken β-actin promoter comprises the nucleic acid sequence of SEQ ID NO:13. 
     
     
         21 . The rAAV vector according to  claim 1 , further comprising at least a first enhancer operably linked to the at least a first nucleic segment, and/or further comprising at least a first mammalian intron sequence operably linked to the at least a first nucleic segment. 
     
     
         22 .- 28 . (canceled) 
     
     
         29 . An isolated mammalian host cell comprising:
 the rAAV vector in accordance with  claim 1 .   
     
     
         30 . (canceled) 
     
     
         31 . A composition comprising:
 (1) the rAAV vector in accordance with  claim 1 ; and   (2) a pharmaceutically-acceptable buffer, carrier, vehicle, or diluent.   
     
     
         32 . (canceled) 
     
     
         33 . The composition according to  claim 31 , formulated for administration to the human eye. 
     
     
         34 .- 36 . (canceled) 
     
     
         37 . A kit comprising:
 (1) the rAAV vector in accordance with  claim 1 ;   (2) instructions for using the component in the diagnosis, prevention, treatment, or amelioration of one or more symptoms of a retinal dystrophy, disease, disorder, or abnormal condition in a human.   
     
     
         38 . The kit according to claim  35 , wherein the retinal dystrophy is diagnosed as Leber congenital amaurosis-1 (LCA-1). 
     
     
         39 .- 40 . (canceled) 
     
     
         41 . A method for preventing, treating or ameliorating a disease, dysfunction, disorder, deficiency, or abnormal condition in a mammal, or one or more symptoms thereof, the method comprising administering to the mammal a composition comprising the rAAV vector in accordance with  claim 1 , in an amount and for a time sufficient to prevent, treat or ameliorate the disease, dysfunction, disorder, deficiency, or abnormal condition in the mammal, or one or more symptoms thereof. 
     
     
         42 .- 43 . (canceled) 
     
     
         44 . The method according to  claim 41 , wherein the mammal is a human neonate, newborn, infant, or juvenile that is at risk for developing or has been diagnosed with a congenital retinal dystrophy such as Leber congenital amaurosis-1 (LCA-1). 
     
     
         45 . A method for providing a mammal with a therapeutically-effective amount of a biologically-active mammalian guanylate cyclase peptide, polypeptide, or protein to a mammal in need thereof, comprising introducing into suitable cells of a mammal in need thereof, an effective amount of the rAAV vector in accordance with  claim 1 , for a time sufficient to produce a biologically-active guanylate cyclase peptide, polypeptide or protein therefrom in at least a first population of cells or at least a first tissue of the mammal into which the rAAV vector has been introduced. 
     
     
         46 .- 56 . (canceled)