METHODS FOR INCREASING THE SIZE OF ANIMALS USING NEEDLELESS DELIVERY CONSTRUCTS
The present invention relates, in part, to methods for increasing the size of a subject by administering a delivery construct comprising growth hormone to a subject. In one aspect, the method for increasing the size of a subject by at least about 12% comprises contacting an apical surface of a polarized epithelial cell of the subject with an amount of a delivery construct comprising growth hormone that is effective to increase the size of the subject by at least about 12%.
1 . A method for increasing the size of a subject by at least about 12%, comprising contacting an apical surface of a polarized epithelial cell of the subject with an amount of a delivery construct effective to increase the size of the subject by at least about 12%, wherein said delivery construct comprises a receptor binding domain, a transcytosis domain, a cleavable linker, and growth hormone (GH), wherein the transcytosis domain transcytoses the GH to and through the basal-lateral membrane of said epithelial cell, and wherein cleavage at said cleavable linker separates said GH from the remainder of said construct, thereby delivering the GH to the subject in an amount effective to increase the size of the subject by at least about 12%.
2 . The method of claim 1 , wherein said receptor binding domain is selected from the group consisting of receptor binding domains from Pseudomonas exotoxin A, cholera toxin, diptheria toxin, shiga toxin, or shiga-like toxin; monoclonal antibodies; polyclonal antibodies; single-chain antibodies; TGF α; EGF; IGF-I; IGF-II; IGF-III; IL-1; IL-2; IL-3; IL-6; MIP-1a; MIP-1b; MCAF; and IL-8.
3 . The method of claim 1 , wherein said receptor binding domain binds to a cell surface receptor selected from the group consisting of α2-macroglobulin receptor, EGFR, IGFR, transferrin receptor, chemokine receptor, CD25, CD11B, CD11C, CD80, CD86, TNFα receptor, TOLL receptor, M-CSF receptor, GM-CSF receptor, scavenger receptor, and VEGF receptor.
4 . The method of claim 1 , wherein said transcytosis domain is selected from the group consisting of transcytosis domains from Pseudomonas exotoxin A, botulinum toxin, diptheria toxin, pertussis toxin, cholera toxin, heat-labile E. coli enterotoxin, shiga toxin, and shiga-like toxin.
5 . The method of claim 1 , wherein said cleavable linker is cleavable by an enzyme that is selected from the group consisting of Cathepsin GI, Chymotrypsin I, Elastase I, Subtilisin AI, Subtilisin AII, Thrombin I, and Urokinase I.
6 . The method of claim 1 , wherein said cleavable linker comprises an amino acid sequence that is selected from the group consisting of Ala-Ala-Pro-Phe (SEQ ID NO.:1), Gly-Gly-Phe (SEQ ID NO.:2), Ala-Ala-Pro-Val (SEQ ID NO.:3), Gly-Gly-Leu (SEQ ID NO.:4), Ala-Ala-Leu (SEQ ID NO.:5), Phe-Val-Arg (SEQ ID NO.:6), Val-Gly-Arg (SEQ ID NO.:7).
7 . The method of claim 1 , wherein the epithelial cell is selected from the group consisting of nasal epithelial cells, oral epithelial cells, intestinal epithelial cells, rectal epithelial cells, vaginal epithelial cells, and pulmonary epithelial cells.
8 . The method of claim 1 , wherein the epithelial cell is a nasal epithelial cell.
9 . The method of claim 1 , wherein the epithelial cell is an intestinal epithelial cell.
10 . The method of claim 1 , wherein said subject is a human.
11 . The method of claim 1 , wherein said delivery construct contacts the apical membrane of the epithelial cell.
12 . The method of claim 1 , wherein said size of said subject is increased by at least about 13%.
13 . The method of claim 1 , wherein said size of said subject is increased by at least about 14%.
14 . The method of claim 1 , wherein said size of said subject is increased by at least about 15%.
15 . The method of claim 1 , wherein said size of said subject is increased by at least about 16%.
16 . The method of claim 1 , wherein said size of said subject is increased by at least about 17%.
17 . The method of claim 1 , wherein said size of said subject is increased by at least about 18%.
18 . The method of claim 1 , wherein said size of said subject is a weight of said subject.
19 . The method of claim 1 , wherein said size of said subject is a length of said subject.
20 . The method of claim 1 , wherein said size of said subject is a height of said subject.
21 . The method of claim 1 , wherein said GH is human growth hormone (hGH).
22 . The method of claim 21 , wherein said hGH has an amino acid sequence that is SEQ ID NO.:8.
23 . The method of claim 1 , further comprising performing the method of claim 1 a second time about 1 day after the method of claim 1 is performed the first time.
24 . The method of claim 1 , further comprising performing the method of claim 1 a second time about 2 days after the method of claim 1 is performed the first time.
25 . The method of claim 1 , further comprising performing the method of claim 1 a second time about 3 days after the method of claim 1 is performed the first time.