Methods and compositions for increasing sialic acid production and treating sialic related disease conditions

Abstract

Disclosed herein are methods of expressing UDP-GlcNAc 2-Epimerase/ManNAc Kinase enzyme (GNE) peptide in a cell of a subject comprising: delivering into the cell of the subject an isolated nucleic acid expression construct that comprises a promoter operatively linked to a nucleic acid sequence encoding a GNE peptide or a therapeutically active fragment thereof, wherein the GNE peptide has the amino acid sequence of SEQ ID NO:3, wherein upon the delivering into the cell of the subject, the nucleic acid expression construct initiates expression of the GNE peptide or a therapeutically active fragment thereof. Also disclosed are methods of producing a GNE peptide in a cell comprising infecting the cell with an isolated nucleic acid construct that comprises a promoter operatively linked to a nucleic acid sequence encoding a GNE peptide or a therapeutically active fragment thereof, wherein the GNE peptide has the amino acid sequence of SEQ ID NO:3.

1 . A method of expressing UDP-GlcNAc 2-Epimerase/ManNAc Kinase enzyme (GNE) peptide in a cell of a subject comprising: delivering into the cell of the subject an isolated nucleic acid expression construct that comprises a promoter operatively linked to a nucleic acid sequence encoding a GNE peptide or a therapeutically active fragment thereof, wherein the GNE peptide has the amino acid sequence of SEQ ID NO:3, wherein upon the delivering into the cell of the subject, the nucleic acid expression construct initiates expression of the GNE peptide or a therapeutically active fragment thereof. 
     
     
         2 . The method of  claim 1 , wherein the isolated nucleic acid expression construct has the sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         3 . The method of  claim 1 , wherein the cell of the subject is in a limb of the subject. 
     
     
         4 . The method of  claim 3 , wherein the delivering step comprises a vascular delivery method in conjunction with a temporary vascular occlusion. 
     
     
         5 . The method of  claim 4 , wherein the delivery method comprises:
 a) infusing fluid into the limb of the subject through an intravascular access, wherein the infused fluid comprises nucleic acid expression construct, and wherein the infused fluid perfuses throughout the limb skeletal muscles or targeted tissue; and   b) applying increased intravascular and intra-tissue pressure by increasing the volume of the infused fluid.   
     
     
         6 . The method of  claim 5 , wherein the nucleic acid expression constructs is delivered in a single dose. 
     
     
         7 . The method of  claim 6 , wherein the single dose comprises a specific concentration of the product irrespective of the total volume being infused, or of a specific molar amount irrespective of the total volume being infused. 
     
     
         8 . The method of  claim 2 , wherein the isolated nucleic acid expression construct further comprises, a transfection-facilitating promoter/enhancer. 
     
     
         9 . The method of  claim 8 , wherein the transfection-facilitating polypeptide comprises an enzymatically active polypeptide able to produce sialic acid. 
     
     
         10 . The method of  claim 9 , wherein the transfection-facilitating polypeptide, or recombinant polypeptide expressed by the isolated nucleic acid expression construct, compromises a hypermorphic form of GNE, producing higher than expected amounts of the sialic acid N-Acetylneuraminate. 
     
     
         11 . The method of  claim 1 , wherein the cell of the subject comprise diploid cells. 
     
     
         12 . The method of  claim 1 , wherein the cell of the subject comprise muscle cells. 
     
     
         13 . The method of  claim 1 , wherein the subject comprises human, ruminant animal, food animal, or work animal. 
     
     
         14 . An isolated nucleic acid molecule the sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         15 . A method of delivering an encoded GNE enzyme comprising:
 a) creating an intravenous access at a point below a knee or an elbow of a limb of a subject;   b) applying a tourniquet at a point proximal to the rest of the body of the subject than the intravenous access point;   c) introducing a single dose of an isolated nucleic acid expression construct into the limb through the intravenous access, wherein the single dose is of sufficient volume to increase intravascular pressure for extravasation of the polynucleotide;   wherein, the isolated nucleic acid construct comprises a promoter operatively linked to a nucleic acid sequence encoding a GNE peptide or a therapeutically active fragment thereof, wherein the GNE peptide has the amino acid sequence of SEQ ID NO:3.   
     
     
         16 . The method of  claim 15 , wherein the isolated nucleic acid construct further comprises a transfection-facilitating polynucleotide. 
     
     
         17 . A method of producing a GNE peptide in a cell comprising infecting the cell with an isolated nucleic acid construct that comprises a promoter operatively linked to a nucleic acid sequence encoding a GNE peptide or a therapeutically active fragment thereof, wherein the GNE peptide has the amino acid sequence of SEQ ID NO:3. 
     
     
         18 . The method of  claim 17 , wherein the cell is a mammalian cell or a bacterial cell. 
     
     
         19 . The method of  claim 17 , wherein the isolated nucleic acid expression construct has the sequence set forth in SEQ ID NO:1 or SEQ ID NO:2.