Modified growth hormone

Abstract

The invention provides, a nucleic acid molecule encoding a growth hormone (GH) in which the RSP sorting signal has been mutated, such that the GH can be constitutively secreted by the nonregulated secretory pathway (NRSP) in a mammalian cell. The invention also provides a nucleic acid molecule encoding a GH in which the three-dimensional conformation of the RSP sorting signal has been altered such that the GH can be constitutively secreted by the NRSP in a mammalian cell.

1 - 35 . (canceled) 
     
     
         36 . A method of treating GHD in a mammal, which method comprises administering to the mammal an isolated and purified nucleic acid molecule encoding GH in which the RSP sorting signal has been mutated such that the GH can be constitutively secreted by the nonregulated secretory pathway (NRSP) in a mammalian cell or a vector comprising the isolated and purified nucleic acid, which expresses an effective amount of the encoded GH, whereupon the GHD in the mammal is treated. 
     
     
         37 . The method of  claim 36 , wherein the vector is administered to the mammal by infusion via the main excretory ducts of the salivary gland of the mammal. 
     
     
         38 . The method of  claim 36 , wherein the isolated and purified nucleic acid molecule encodes the amino acid sequence of SEQ ID NO: 2 in which the sorting signal comprises Glu 174, Leu 177, Val 185 and Glu 186 and one or more of the codons encoding the aforementioned amino acids is mutated or a vector comprising the isolated and purified nucleic acid, which expresses an effective amount of the encoded GH, whereupon the GHD in the mammal is treated. 
     
     
         39 . The method of  claim 38 , wherein the vector is administered to the mammal by infusion via the main excretory ducts of the salivary gland of the mammal. 
     
     
         40 . A method of treating GHD in a mammal, which method comprises administering to the mammal an isolated and purified nucleic acid molecule encoding GH in which the three-dimensional conformation of the RSP sorting signal has been altered such that the GH can be constitutively secreted by the NRSP in a mammalian cell or a vector comprising the isolated and purified nucleic acid, which expresses an effective amount of the encoded GH, whereupon the GHD in the mammal is treated. 
     
     
         41 . The method of  claim 40 , wherein the vector is administered to the mammal by infusion via the main excretory ducts of the salivary gland of the mammal. 
     
     
         42 . A method of treating GHD in a mammal, which method comprises administering to the mammal host cells which have been contacted ex vivo with an isolated and purified nucleic acid molecule encoding GH in which the RSP sorting signal has been mutated such that the GH can be constitutively secreted by the nonregulated secretory pathway (NRSP) in a mammalian cell or a vector comprising the isolated and purified nucleic acid, which expresses an effective amount of the encoded GH, whereupon the GHD in the mammal is treated. 
     
     
         43 . The method of  claim 42 , wherein the host cells are autologous to the mammal. 
     
     
         44 . The method of  claim 43 , wherein the host cells are derived from a biopsy of secretory gland tissue. 
     
     
         45 . The method of  claim 44 , wherein the secretory gland tissue is salivary gland tissue. 
     
     
         46 . The method of  claim 42 , wherein the isolated and purified nucleic acid molecule of  claim 36 , which encodes the amino acid sequence of SEQ ID NO: 2 in which the sorting signal comprises Glu 174, Leu 177, Val 185 and Glu 186 and one or more of the codons encoding the aforementioned amino acids is mutated or a vector comprising the isolated and purified nucleic acid, which expresses an effective amount of the encoded GH, whereupon the GHD in the mammal is treated. 
     
     
         47 . The method of  claim 46 , wherein the host cells are autologous to the mammal. 
     
     
         48 . The method of  claim 47 , wherein the host cells are derived from a biopsy of secretory gland tissue. 
     
     
         49 . The method of  claim 48 , wherein the secretory gland tissue is salivary gland tissue. 
     
     
         50 . A method of treating GHD in a mammal, which method comprises administering to the mammal host cells which have been contacted ex vivo with an isolated and purified nucleic acid molecule encoding GH in which the three-dimensional conformation of the RSP sorting signal has been altered such that the GH can be constitutively secreted by the NRSP in a mammalian cell or a vector comprising the isolated and purified nucleic acid, which expresses an effective amount of the encoded GH, whereupon the GHD in the mammal is treated. 
     
     
         51 . The method of  claim 50 , wherein the host cells are autologous to the mammal. 
     
     
         52 . The method of  claim 51 , wherein the host cells are derived from a biopsy of secretory gland tissue. 
     
     
         53 . The method of  claim 52 , wherein the secretory gland tissue is salivary gland tissue. 
     
     
         54 . A method making a GH in which the RSP sorting signal is mutated, which method comprises mutating one or more amino acids in the RSP sorting signal in GH, whereupon a GH in which the RSP sorting signal is mutated is obtained. 
     
     
         55 . A GH with a mutated sorting signal obtained in accordance with  claim 54 . 
     
     
         56 . A method making a GH in which the three-dimensional conformation of the RSP sorting signal is altered, which method comprises mutating one or more amino acids outside of the RSP sorting signal so that the three-dimensional conformation of the amino acids of the RSP sorting signal in GH is altered, whereupon a GH with an RSP having an altered three-dimensional conformation is obtained. 
     
     
         57 . A GH with an RSP sorting signal having an altered three-dimensional conformation obtained in accordance with  claim 56 . 
     
     
         58 . A method of making a nucleic acid molecule encoding a GH that can be constitutively secreted by the NRSP in a mammalian cell, which method comprises mutating one or more codons encoding amino acids in the RSP sorting signal in an isolated and purified nucleic acid molecule encoding GH such that, upon expression in a mammalian cell, the GH can be constitutively secreted by the NRSP in the mammalian cell. 
     
     
         59 . A nucleic acid molecule encoding a GH with a mutated RSP sorting signal obtained in accordance with the method of  claim 58 . 
     
     
         60 . A method of making a nucleic acid molecule encoding a GH that can be constitutively secreted by the NRSP in a mammalian cell, which method comprises mutating one or more codons encoding amino acids outside of the RSP sorting signal in an isolated and purified nucleic acid molecule encoding GH such that, the three-dimensional conformation of the amino acids of the RSP sorting signal in GH is altered and, upon expression in a mammalian cell, the GH can be constitutively secreted by the NRSP in the mammalian cell. 
     
     
         61 . A nucleic acid molecule encoding a GH with an RSP sorting signal having an altered three-dimensional conformation obtained in accordance with the method of  claim 60 .