Anti-inflammatory therapies using cytokine signaling regulated by a SOCS-3 promoter

Abstract

Disclosed is a nucleic acid construct comprising a murine SOCS-3 promoter sequence having SEQ. ID. NO.: 1, or a non-murine homologue thereof, or an operative fragment or derivative. The construct can also contain, operatively linked to the SOCS-3 promoter, a gene encoding any preselected protein, and optionally contains a reporter gene to facilitate detection and/or selection of successfully transfected cells. Also disclosed are a transgenic vertebrate cell containing the nucleic acid construct and transgenic non-human vertebrates comprising such cells. The nucleic acid construct is useful in methods of treating a growth retardation or growth acceleration disorder in a human subject and in a method of treating an autoimmune disease, immune disease, or inflammatory condition in a human subject. A kit for genetically modifying a vertebrate cell includes a polynucleotide comprising the murine SOCS-3 promoter sequence is also disclosed.

We claim:  
     
         1 . A method of treating an autoimmune disease, immune disease, or an inflammatory condition in a human subject, comprising: 
 genetically modifying a gp130-responsive cell of a human subject having an autoimmune disease, immune disease, or inflammatory condition with a nucleic acid construct comprising a SOCS-3 promoter sequence, or functional fragment thereof, operatively linked, in a transcriptional unit, to a DNA sequence encoding a functional anti-inflammatory cytokine of the gp130 subunit signaling cytokine family linked to a functional secretory signal; and    causing the cell, in vivo, to transcribe from said transcriptional unit, in response to a pro-inflammatory cytokine of the gp130 signaling subunit cytokine family, a functional mRNA transcript, whereby the anti-inflammatory cytokine is produced and secreted in response to the pro-inflammatory cytokine, and one or more symptoms of the autoimmune disease, immune disease, or inflammatory condition is improved.    
     
     
         2 . The method of  claim 1 , wherein the cell is a pituitary, adrenal, hypothalamic, intestinal, kidney, liver, immune-competent, or bone-forming cell.  
     
     
         3 . The method of  claim 1 , wherein the cell is a hepatocyte, lymphocyte, osteoblast, neuron, intestinal epithelial, corticotroph, somatotroph, lactotroph, or gonadotroph cell.  
     
     
         4 . The method of  claim 1 , wherein the cell is a cell derived from a pituitary tumor, adrenal tumor, hypothalamic tumor, or liver tumor.  
     
     
         5 . The method of  claim 1 , wherein the SOCS-3 promoter sequence is a murine SOCS-3 promoter having a nucleotide sequence (SEQ. ID. NO: 1), or an operative fragment thereof, or an operative derivative of either of these, said derivative being selected from the group consisting essentially of a mutation, a polymorphism, and a variant allele with respect to any nucleotide position of SEQ. ID. NO.: 1, and said derivative being a derivative that does not fully eliminate promoter activity.  
     
     
         6 . The method of  claim 1 , wherein said transcriptional unit further comprises a DNA sequence encoding a reporter protein.  
     
     
         7 . The method of  claim 6 , wherein the encoded reporter protein is a fluorescent or light-emitting protein.  
     
     
         8 . The method of  claim 7 , wherein the encoded fluorescent or light-emitting protein is a green fluorescent protein, yellow fluorescent protein, blue fluorescent protein, phycobiliprotein, luciferase, or apoaequorin.  
     
     
         9 . The method of  claim 1 , further comprising at least one insulator element flanking said transcriptional unit.  
     
     
         10 . The method of  claim 9 , wherein at least one of said insulator element(s) is a chicken β-globin insulator element.  
     
     
         11 . The method of  claim 1 , wherein the pro-inflammatory cytokine is IL-6 or LIF.  
     
     
         12 . The method of  claim 1 , wherein the anti-inflammatory cytokine is IL-11.  
     
     
         13 . The method of  claim 1 , wherein the autoimmune disease, immune disease, or inflammatory condition is Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, Grave's disease, or a neuroendocrinological response to stress.  
     
     
         14 . The method of  claim 1 , further comprising obtaining the gp130-responsive cell from a tissue of the human subject; genetically modifying the cell in vitro; and re-implanting the genetically modified cell into the tissue of the human subject.  
     
     
         15 . A method of treating an autoimmune disease, immune disease, or inflammatory condition in a human subject, comprising: 
 obtaining a gp130-responsive cell from a tissue of a human subject having an autoimmune disease, immune disease, or inflammatory condition;    transfecting said cell with a nucleic acid construct comprising a murine SOCS-3 promoter having a nucleotide sequence (SEQ. ID. NO: 1), or an operative fragment thereof, or an operative derivative of either of these, operatively linked, in a transcriptional unit, to a DNA sequence encoding a functional anti-inflammatory cytokine of the gp130 subunit signaling cytokine family linked to a functional secretory signal; and    re-implanting said cell within the tissue of said human subject, whereby the anti-inflammatory cytokine is produced and secreted, in vivo, in response to a pro-inflammatory cytokine of the gp130 subunit signaling cytokine family, and one or more symptoms of the autoimmune disease, immune disease, or inflammatory condition is improved.    
     
     
         16 . The method of  claim 15 , wherein the cell is a pituitary, adrenal, hypothalamic, intestinal, kidney, liver, immune-competent, or bone-forming cell.  
     
     
         17 . The method of  claim 15 , wherein the cell is a hepatocyte, lymphocyte, osteoblast, intestinal epithelial, corticotroph, somatotroph, lactotroph, or gonadotroph cell.  
     
     
         18 . The method of  claim 15 , wherein the cell is a cell derived from a pituitary tumor, adrenal tumor, hypothalamic tumor, or liver tumor.  
     
     
         19 . The method of  claim 15 , wherein said transcriptional unit further comprises a DNA sequence encoding a reporter protein.  
     
     
         20 . The method of  claim 19 , wherein the encoded reporter protein is a fluorescent or light-emitting protein.  
     
     
         21 . The method of  claim 20 , wherein the fluorescent or light-emitting protein is a green fluorescent protein, yellow fluorescent protein, blue fluorescent protein, phycobiliprotein, luciferase, or apoaequorin.  
     
     
         22 . The method of  claim 15 , further comprising at least one insulator element flanking said transcriptional unit.  
     
     
         23 . The method of  claim 22 , wherein at least one of said insulator element(s) is a chicken β-globin insulator element.  
     
     
         24 . The method of  claim 15 , wherein the anti-inflammatory cytokine is IL-11.  
     
     
         25 . The method of  claim 15 , wherein the pro-inflammatory cytokine is IL-6 or LIF.  
     
     
         26 . The method of  claim 15 , wherein the autoimmune disease, immune disease, or inflammatory condition is Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, Grave's disease, an allergic or anaphylactic reaction, or a neuroendocrinological response to stress.