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Matrix-targeted fusion polypeptides for tissue regeneration and wound healingMatrix-targeted fusion polypeptides for tissue regeneration and wound healing description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20090093407, Matrix-targeted fusion polypeptides for tissue regeneration and wound healing. Brief Patent Description - Full Patent Description - Patent Application Claims
This application is a continuation application of and claims priority to U.S. application Ser. No. 10/733,852, filed Dec. 10, 2003, which is a continuation application of U.S. Ser. No. 09/624,874, filed Jul. 21, 2000, which claims priority to U.S. Provisional Application Ser. No. 60/145,488, filed Jul. 21, 1999. The disclosure of these prior applications are considered part of (and are incorporated by reference in) the disclosure of this application. The present invention relates generally to pharmaceutical agents targeted to a tissue for promoting tissue regeneration, and more particularly to the use of collagen-binding domains to target a growth factor to a desired tissue. Impaired tissue healing is a significant problem in health care. Chronic, non-healing wounds are a major cause of prolonged morbidity in the aged human population. Tissue repair is particularly relevant to bedridden or diabetic patients who develop severe, external, non-healing skin ulcers. In addition, those patients suffering from internal lesions, such as those associated with disorders of the digestive tract, are particularly susceptible to the effects of non or slow-healing tissue damage. Pharmaceutical agents that promote tissue regeneration at the site of a lesion, such as growth factors, have been utilized to accelerate wound repair. Growth factors are molecules that function not only as growth simulators (mitogens), but also as growth inhibitors. Growth factors are also known to stimulate cell migration (e.g., mitogenic cytokines), function as chemotactic agents, inhibit cell migration or invasion of tumor cells, modulate differentiated functions of cells, be involved in apoptosis, and promote survival of cells. For example, epidermal growth factor (EGF) is a mitogen that not only effects suppression of gastric acid secretion and fetal lung development, but also effects wound healing and epidermal regeneration (Franklin et al., J. Lab. Clin. Med., 108:103, 1985). EGF has been shown to be a potent stimulator of epithelial cell proliferation in the human intestine (Alison et al., Cell Biol. Int., 18:1, 1994) and other tissues. Previous studies have demonstrated the viability of targeting extracellular matrix molecules, such as exposed collagen, for delivery of a pharmaceutical agent to a specific tissue. For example, von Willebrand Factor (vWF)-derived collagen-binding domains have been used to target the TGF-β family of growth factors to damaged tissue (Tuan et al., Conn. Tiss. Res., 34:1, 1996; Han et al., Protein Expr. Purif., 11:169, 1997). Pharmaceutical agents that promote tissue regeneration are useful not only for treating disorders associated with impaired tissue regeneration, but also for promoting tissue regeneration associated with surgical procedures, for example. Several strategies have been developed to accomplish enhanced tissue repair for the treatment of damaged tissue. Within these strategies, there is a need for the controlled, sustained, site-specific targeting of a pharmaceutical agent to a wound site for the purpose of promoting tissue regeneration. The present invention provides new compositions and methods to induce therapeutic repair of epithelial tissue by specifically targeting tissue in need of such repair with a fusion polypeptide of the invention. The invention promotes localized wound healing by providing a cell proliferation-modulating agent fused to a collagen-binding domain. The new compositions and methods are useful for wound healing in general and for promoting repair of colonic lesions, for the capture and expansion of crypt stem cells, and the enhancement of retroviral gene transfer in colonic mucosal cells, in particular. In one embodiment, a fusion polypeptide comprising an epithelial cell proliferation-modulating agent and a collagen-binding domain capable of binding to collagen, is provided. A nucleic acid sequence encoding the fusion polypeptide is also provided. In another aspect, the invention provides a method of producing a fusion polypeptide of the invention having a collagen-binding domain and an epithelial cell proliferation-modulating agent, by growing the host cells containing a nucleic acid encoding the fusion polypeptide under conditions that allow expression of the nucleic acid sequence and recovering the fusion polypeptide. In another embodiment, the invention provides method for modulating epithelial cell proliferation in a subject. In one aspect, the modulating is by administering to the subject a fusion polypeptide comprising a collagen-binding domain source linked to an epithelial cell proliferation-modulating agent. In another aspect, the modulating is by administering to the subject a therapeutically effective amount of a nucleic acid sequence encoding a fusion polypeptide comprising a collagen-binding domain linked to an epithelial cell proliferation-modulating agent. In a further embodiment, a tissue graft, comprising isolated tissue comprising epithelial cells treated with a fusion polypeptide comprising a collagen-binding domain linked to an epithelial cell proliferation-modulating agent, is provided. Also provided is a method of preparing a tissue graft using an effective amount of a fusion polypeptide of the invention. The invention further provides a method for modulating tissue regeneration in a subject. In one aspect, the method comprises administering to the subject a therapeutically effective amount of a fusion polypeptide of the invention. In another aspect, the method comprises administering to the subject a therapeutically effective amount of a nucleic acid encoding a fusion polypeptide of the invention. In another embodiment, the invention provides a pharmaceutical composition for promoting tissue repair. In one aspect, the composition comprises a fusion polypeptide comprising a collagen-binding domain linked to an epithelial cell proliferation-modulating agent in a pharmaceutically acceptable carrier. In another aspect, the composition comprises a nucleic acid encoding a fusion polypeptide of the invention. The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. Continue reading about Matrix-targeted fusion polypeptides for tissue regeneration and wound healing... 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