SBIR-STTR Award

Exposure of human skin to ultraviolet light (UV) triggers a progression of events that is initiated by DNA damage and immune suppression and can ultimately result in mutagenesis, cell proliferation, actinic keratoses, and skin cancers. The occurrence of these diseases is rapidly increasing, affecting over 1 million people in the United States annually. The carcinogenic effects of UV light are directly mediated by dipyrimidine DNA photoproducts. Human cells have only one mechanism to repair these DNA lesions, while lower organisms possess multiple pathways to repair the deleterious consequences of UV- induced DNA damage. In order to implement proactive strategies to treat and prevent several human diseases, including skin cancer, that can be directly attributed to inefficient repair of UV-induced DNA lesions, the founders of Restoration Genetics, Inc. have discovered, characterized and patented multiple DNA repair enzymes that possess catalytic activities that can initiate the BER pathway in human cells. These highly stable, patented enzymes have been engineered to specifically localize to the nucleus, thus significantly increasing the repair efficiency. Pilot studies have demonstrated the feasibility of incorporating these enzymes in an active form in liposomes that, upon topical application to skin, enhance DNA repair of UV light-induced DNA damage. In order to maximize efficacy of these enzymes to accelerate DNA repair in reconstituted human skin, the following aims are proposed: 1) Purify and encapsulate the nuclear forms of the patented enzymes in a variety of liposomal targeting vehicles that are used for topical skin delivery; 2) Test the efficacy of the enzymes using fully-differentiated human skin culture in multiple short-term assays. The commercial market potential for this technology is diverse and will include pharmaceutical applications for the prevention of skin cancers. The ultimate goal of these investigations is to reduce UV-induced skin cancers and immunosuppression through topical delivery of high specific activity DNA repair enzymes that localize to critical subcellular organelles. Acute and chronic exposure to sunlight is a primary risk factor for skin cancer. Human cells possess only one mechanism for the repair of UV-induced DNA damage. Our laboratories have patented new enzymes that activate a second DNA repair pathway. Thus, our technology will augment and dramatically improve human DNA repair capacity for removal of sunlight-induced damage. The anticipated societal benefits are to significantly reduce the number and average age of onset of new skin cancers

Current health care modalities for humans diagnosed with either precancerous actinic keratoses or skin cancers (including basal and squamous cell carcinomas) primarily involve surgical excision of the lesions or chemical or photodynamic burning of the affected tissues. These post-diagnostic treatments are directed to disease management, while disease prevention strategies focus on public education that advocates for limiting sunlight exposures, especially in the first two decades of life. However, with ~1.4 million new cases of skin cancer diagnosed in the United States each year, additional prevention strategies are needed. To address this need, Restoration Genetics, Inc (RGI) has developed and patented technologies activating a second, nuclear- targeted DNA repair pathway, base excision repair, for the removal of the primary DNA lesions caused by sunlight exposure, cyclobutane pyrimidine dimers (CPDs). Using Phase I STTR funding, RGI successfully demonstrated the purification of the critical repair enzyme, chlorella virus pyrimidine dimer DNA glycosylase (Cv-pdg-NLS, RG-101), encapsulation of the enzyme into liposomes, and the successful initiation of rapid repair of CPDs in a human skin model. Building from these data, in collaboration with Oregon Health &Science University, RGI will carry out a series of preclinical studies including cGMP process development, biological efficacy analyses, and commercialization development, all of which culminate in a pre-IND meeting with the FDA. Specifically, RG-101 will be produced and packaged in a large-scale cGMP facility at the University of Nebraska and RG-101 encapsulated in liposomes will be tested for preclinical biological efficacy to 1) activate repair of solar-simulated light in a human skin model, 2) prevent UV-induced carcinogenesis in a rodent model, and 3) reduce tumor frequencies following an initial tumor diagnosis. Additionally, pharmacokinetic and toxicology studies will be designed using rabbit and pig models. These investigations complement well the commercialization plan that targets three markets: 1) individuals who are deficient in the normal DNA repair mechanism for CPD removal (conducted under Orphan Drug Status), 2) organ transplant patients who are highly susceptible to frequent, highly aggressive skin cancers, and 3) individuals with high frequency actinic keratoses and cancers, as well as the general public. Collectively, the data will be presented to the FDA in a pre-IND meeting for guidance in pharmacology and toxicology trials and Phase I human clinical trials. Specifically, in this meeting we will present manufacturing, characterization and encapsulation data, biochemical activities of the enzyme and in vitro efficacy in fully differentiated human skin models, mouse carcinogenesis data, and the design of animal pharmacology and toxicology assessments.

Public Health Relevance:
Acute and chronic exposures to sunlight are the primary risk factors for the development of skin cancer. Human cells possess only one mechanism for the repair of UV-induced DNA damage. Our laboratories have patented new enzymes that activate a second DNA repair pathway. Thus, our technology will augment and dramatically improve human DNA repair capacity for removal of sunlight-induced damage. The anticipated societal benefits are to significantly reduce the number and average age of onset of new skin cancers.

Thesaurus Terms:
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