Methods and compositions for regulation of metabolic disorders

Abstract

Cells and methods of using these cells for expressing a transgene expressing a protein that is aberrantly expressed in a metabolic disorders from a safe harbor locus.

What is claimed is: 
     
         1 . A method of expressing a protein product in a subject with a metabolic disorder, the method comprising:
 administering an isolated cell to the subject, the isolated cell comprising an exogenous transgene encoding a protein, wherein the exogenous transgene is integrated site-specifically using a zinc finger nuclease into an endogenous safe harbor gene and further wherein the protein is a functional version of a protein that is aberrantly expressed in the metabolic disorder and further wherein the metabolic disorder is selected from the group consisting of methylmalonic acidemia, propionic acidemia, glycogen storage diseases type 1, familial hypercholesterolemia (FH) is a common genetic and metabolic disease, urea cycle disorders, Crigler Najjar Syndrome (CNS), Gilbert syndrome, hepatorenal tyrosinemia, primary hyperoxaluria. transthyretin gene (TTR)-mediated amyloidosis (ATTR), Wilson's disease, phenylketonuria (PKU), and familial lipoprotein lipase deficiency (LPLD).   
     
     
         2 . The method of  claim 1 , wherein the cell is selected from the group consisting of a hepatic cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem cell, a hepatic cell, a hepatic stem cell, and a red blood precursor cell. 
     
     
         3 . The method of  claim 1 , wherein the endogenous safe harbor gene is selected from the group consisting of CCR5, HPRT, AAVS1, Rosa and albumin. 
     
     
         4 . The method of  claim 1 , wherein the transgene encodes a protein selected from the group consisting of MMAA, MMAB, MMACHC, MMADHC (C2orf25), MTRR, LMBRD1, MTR, propionyl-CoA carboxylase (PCCA or PCCB), a glucose-6-phosphate transporter (G6PT) protein or glucose-6-phosphatase (G6Pase), an LDL receptor (LDLR), an ApoB protein, an LDLRAP-1 protein, a PCSK9 protein, a mitochondrial protein such as NAGS (N-acetylglutamate synthetase), CPS1 (carbamoyl phosphate synthetase I), and OTC (ornithine transcarbamylase), ASS (argininosuccinic acid synthetase), ASL (argininosuccinase acid lyase) and/or ARG1 (arginase), and/or a solute carrier family 25 (SLC25A13, an aspartate/glutamate carrier) protein, a UGT1A1 or UDP glucuronsyltransferase polypeptide A1, a fumarylacetoacetate hydrolyase (FAH), an alanine-glyoxylate aminotransferase (AGXT) protein, a glyoxylate reductase/hydroxypyruvate reductase (GRHPR) protein, a transthyretin gene (TTR) protein, an ATP7B protein, a phenylalanine hydroxylase (PAH) protein and a lipoprotein lyase (LPL) protein. 
     
     
         5 . The method of  claim 1 , wherein expression of the transgene is driven by an endogenous promoter. 
     
     
         6 . The method of  claim 1 , wherein the transgene is operably linked to a promoter sequence that drives expression of the transgene. 
     
     
         7 . The method of claim of  claim 1 , wherein the nuclease is administered to the cell via a vector, an mRNA or as an intact protein.