# US 2013/0158103 A1 · Method of Tissue-Selective Targeted Gene Transfer > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2013/0158103 A1 - Document ID: US20130158103A1 - Status: Published application - Kind: A1 - Application number: 13/683,944 - Filed: 2012-11-21 - Published: 2013-06-20 - Earliest claimed: 2011-11-23 - Assignees: The Curators of the University of Missouri - Inventors: Rajiv R. Mohan - Web page: https://patentgrep.com/patent/US20130158103A1 ## CPC - A61K 48/0075: {characterised by an aspect of the delivery route, e.g. oral, subcutaneous} - A61K 38/1709: {from mammals} - A61K 38/39: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG] - C12N 2750/14143: viral genome or elements thereof as genetic vector - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to methods of delivering a gene, such as a therapeutic gene, to a desired area of stroma of a cornea that involves removing the corneal epithelium and dehydrating the cornea. Certain aspects of the present invention relate to methods of treating corneal scarring by delivering a TGFβ-antagonizing gene packaged in a viral vector. ## Claims ### Claim 1 A method of delivering a gene to a desired area of stroma of a cornea, the method comprising the steps of: (a) preparing the cornea by: (i) removing corneal tissue to expose at least a portion of the corneal stroma, and (ii) dehydrating the exposed portion of the stroma; (b) applying a viral vector that comprises the gene to the dehydrated portion of the stroma. ### Claim 2 (depends on 1) The method of claim 1 further comprising the step of removing excess viral vector after application in step (b). ### Claim 3 (depends on 1) The method of claim 1 wherein the viral vector is an AAV vector in solution with a titer of from about 1×108 vg/ml to about 6.5×1012 vg/ml. ### Claim 4 (depends on 1) The method of claim 1 wherein the viral vector is in solution and is applied in a volume of solution of from about 1 μl to about 100 μl. ### Claim 5 (depends on 1) The method of claim 1 wherein the viral vector is an AAV vector in solution with a titer of from about 1×108 vg/ml to about 6.5×1012 vg/ml and the AAV vector is applied in a volume of solution of from about 1 μl to about 100 μl. ### Claim 6 (depends on 1) The method of claim 1 wherein removing the corneal tissue in (a)(i) comprises mechanical scraping. ### Claim 7 (depends on 1) The method of claim 1 wherein dehydrating the exposed portion of the stroma in (a)(ii) comprises contacting the surface of the cornea with a flow of air at a temperature in the range of from about 40° C. to about 45° C. for a total duration of from about 10 seconds to about 60 seconds, wherein said flow is at a rate in the range of about 6 meters per second to about 10 meters per second. ### Claim 8 (depends on 1) The method of claim 1 wherein following preparation of the cornea in (a), a physical barrier is placed on the cornea that encompasses the desired surface area of stroma of the cornea to which the gene is to be delivered, and wherein the viral vector applied in (b) is in a solution that is applied to the area encompassed by the physical barrier. ### Claim 9 A method of treating corneal scarring comprising applying a viral vector that comprises a TGFβ-antagonizing gene to the stroma of a cornea. ### Claim 10 (depends on 9) The method of claim 9 wherein the viral vector is an AAV vector. ### Claim 11 (depends on 9) The method of claim 9 wherein the TGFβ-antagonizing gene is decorin. ### Claim 12 (depends on 9) The method of claim 9 wherein the TGFβ-antagonizing gene is delivered to a desired area of stroma of the cornea, the method comprising the steps of: (a) preparing the cornea by: (i) removing corneal tissue to expose at least a portion of the corneal stroma, and (ii) dehydrating the exposed portion of the stroma; (b) applying an AAV viral vector comprising the TGFβ-antagonizing gene to the dehydrated portion of the stroma. ### Claim 13 (depends on 12) The method of claim 12 wherein the AAV vector is in solution with a titer of from about 1×108 vg/ml to about 6.5×1012 vg/ml. ### Claim 14 (depends on 12) The method of claim 12 wherein the viral vector is in solution and is applied in a volume of solution of from about 1 μl to about 100 μl. ### Claim 15 (depends on 12) The method of claim 12 further comprising the step of removing excess AAV vector after application in step (b). ### Claim 16 (depends on 12) The method of claim 12 wherein the TGFβ-antagonizing gene is decorin. ### Claim 17 (depends on 12) The method of claim 12 wherein removing the corneal tissue in (a)(i) comprises mechanical scraping. ### Claim 18 (depends on 12) The method of claim 12 wherein dehydrating the exposed portion of the stroma in (a)(ii) comprises contacting the surface of cornea with a flow of air at a temperature in the range of from about 40° C. to about 45° C. for a total duration of from about 10 seconds to about 60 seconds, wherein said flow is at a rate in the range of about 6 meters per second to about 10 meters per second. ### Claim 19 (depends on 12) The method of claim 12 wherein following preparation of the cornea in (a), a physical barrier is placed on the cornea that encompasses the desired surface area of stroma of the cornea to which the gene is to be delivered, and wherein the viral vector applied in (b) is in a solution that is applied to the area encompassed by the physical barrier. ### Claim 20 A method of treating corneal scarring comprising applying an AAV5 vector that comprises a decorin gene to the stroma of a cornea wherein the gene is delivered to a desired area of stroma of a cornea, the method further comprising the steps of: (a) preparing the cornea by: (i) removing corneal tissue by mechanical scraping to expose at least a portion of the corneal stroma, and (ii) dehydrating the exposed portion of the stroma; (b) applying the AAV5 vector that comprises the decorin gene to the dehydrated portion of the stroma, wherein the AAV5 vector is in solution with a titer of from about 1×108 vg/ml to about 6.5×1012 vg/ml and is applied in a volume of solution of from about 1 μl to about 100 μl, wherein dehydrating the exposed portion of the stroma in (a)(ii) comprises contacting the surface of the cornea with a flow of air at a temperature in the range of from about 40° C. to about 45° C. for a total duration of about 10 seconds to about 60 seconds, wherein said flow is at a rate in the range of about 6 meters per second to about 10 meters per second, and wherein following preparation of the cornea in (a), a physical barrier is placed on the cornea that encompasses the desired surface area of stroma of the cornea to which the gene is to be delivered, and wherein the viral vector applied in (b) is applied to the area encompassed by the physical barrier.