Anti-scarring ribozymes and methods
View Patent ↗Methods and compositions for treating scarring conditions associated with increased expression of connective tissue growth factor (CTGF). Aspects of the invention include ribozymes that cleave mRNA targets required for CTGF expression, cells containing anti-CTGF ribozymes and vectored anti-CTGF ribozymes suitable for delivery to cellular targets capable of CTGF expression.
1. A ribozyme that specifically cleaves a target RNA sequence encoded by a CTGF gene, said ribozyme encoded by a nucleic acid comprising a sequence of SEQ ID NO:48.
2. The ribozyme of claim 1 , wherein the ribozyme is present in a hammerhead configuration.
3. The ribozyme of claim 2 , wherein the ribozyme specifically cleaves target RNA sequences encoded by CTGF nucleotide sequence SEQ ID NO:50.
4. An expression vector comprising an isolated nucleic acid encoding a CTGF ribozyme of SEQ ID NO:48.
5. The expression vector of claim 4 , wherein said vector is a plasmid.
6. The expression vector of claim 5 , wherein the plasmid directs synthesis of a self-cleaving hairpin ribozyme attached to a CTGF ribozyme in a hammerhead configuration.
7. The expression vector of claim 6 , wherein the plasmid is pTRUF21.
8. A method for reducing CTGF mRNA or protein in a cell, the method comprising the steps of: (a) group providing a tissue comprising a cell expressing a target RNA sequence encoded by a CTGF gene;and (b) contacting said tissue with a vector comprising a nucleic acid that encodes at least one ribozyme that specifically cleaves a target RNA sequence encoded by a CTGF gene, in an amount effective to reduce said CTGF MRNA or protein expression in said cell, wherein said at least one ribozyme is encoded by a nucleic acid comprising a sequence of SEQ ID NO:48 or a complement of SEQ ID NO:48.
9. The method of claim 8 , wherein said cell is a fibroblast.
10. The method of claim 8 , further comprising contacting said tissue with said vector in a subject having or at risk for developing a condition causing a scar in said tissue.
11. The method of claim 10 , wherein said condition is a fibrotic disorder.
12. The method of claim 11 , wherein said fibrotic disorder is selected from the group consisting of scleroderma, keloids, liver cirrhosis, kidney fibrosis, peritoneal adhesions, tendon adhesions, breast implant capsule adhesions, burn scars spinal cord injuries, bile duct atresia, subepithelial firbrosis, fibrous dysplasia, and tympanic membrane fibrosis.
13. The method of claim 11 , wherein said condition is wound healing following surgery.
14. The method of claim 13 , wherein said surgery is corneal surgery.
15. The method of claim 13 , wherein said surgery is glaucoma filtering surgery.
16. The method of claim 8 , wherein said tissue is an ocular tissue selected from the group consisting of cornea, conjunctiva, sciera and trabecular meshwork.