SBIR-STTR Award

STTR 608040: Optimization of the UValidate platform to measure genotoxicity associated with current problematic UV chemical blockers Optimization of the UValidate™ platform to measure genotoxicity associated with current problematic UV chemical blockers. / (word count: 375) Over-the-counter active ingredients that block UVA and UVB are required in all sunscreens, as mandated by the FDA. Recently the safety of these active ingredients has come under scrutiny both from a human health and ecological perspective. Yet there is a lack of alternative UV blockers available to manufacturers leaving the public in a precarious position. The objective of this Phase I STTR is to calibrate a novel UV active ingredient testing platform (termed UValidate™) in order to test potential UV chemical blockers for the skin care industry. Specifically, these chemical blockers would be required to reduce or prevent the deleterious effects of UV irradiation in human cells that comprise the bulk of the epidermis, including fibroblasts, keratinocytes, and melanocytes. Biological endpoints that will be measured after sunscreen and UV exposure include: DNA damage associated with oxidative stress, direct UV DNA damage products (CPD/6-4PP), modulation of DNA repair capacity, and mutagenicity. The product would initially (Phase I) be a comprehensive sunscreen active ingredient testing platform, that in Phase II would be further expanded to identify new or repurposed UV-protective compounds for downstream clinical evaluation. Technical milestones that will be completed in Phase I include: 1) Integrating a novel UV DNA damage-detecting protein-probe into the CometChip methodology, 2) Configuring UVR micro-LED technology to provide a novel UV irradiation source adaptable to screening platforms. 3) Developing a data pipeline that is able to autonomously take the large amounts of information accumulated by the UValidate system and transform the data into a compound-specific predictive result. UValidate is designed to complement rather than replace current sunscreen testing regimes by measuring biological endpoints not evaluated by classical UV absorption measurements. Once we have calibrated UValidate by comprehensively investigating the current OTC UV chemical blockers, we will use data-driven learning protocols to develop a system that can make predictive results about new compounds (Phase II) and in the future be expanded to test the combinatory effects of other agents with sunscreen active ingredients, focusing specifically on insect repellents, moisturizers and environmental pollutants. The impact of the platform will be immediate, with data from the current UV chemical blockers becoming part of the non-clinical assessment that will be used by the FDA to assess the future of the compounds in the marketplace.

Public Health Relevance Statement:
STTR 608040: Optimization of the UValidate platform to measure genotoxicity associated with current problematic UV chemical blockers NARRATIVE Over-the-counter (OTC) active ingredients that block UVA and UVB are required in all sunscreens, as mandated by the FDA. Yet the safety of these compounds has recently come under scrutiny both from a human health and ecological perspective. The objective of this Phase I STTR is to calibrate a novel UV active ingredient testing platform (termed UValidate™), in order to test current (Phase I), and novel (Phase II) chemical UV blockers for the skin care industry. Specifically, these chemical blockers would be required to reduced or prevent the deleterious effects of UV irradiation, induced by micro-LED technology, using human cell types that comprise the epidermis. The impact of the platform will be immediate, with data from the OTC chemical blockers becoming part of the non-clinical assessment that will be used by the FDA to assess the future of the chemical blockers in the marketplace.

Project Terms:
absorption; Biological Assay; Assay; Bioassay; Biologic Assays; Cells; Cell Body; Complement; Complement Proteins; Cosmetics; cosmetic product; DNA Damage; DNA Injury; DNA Repair; DNA Damage Repair; Unscheduled DNA Synthesis; Environmental Pollutants; Epidermis; Fibroblasts; Future; Health; Human; Modern Man; Industry; Insect Repellents; keratinocyte; Kinetics; Learning; melanocyte; Methodology; Proteins; Risk; Safety; Sunscreening Agents; Sunblock; Sunscreens; Technology; Testing; Measures; Skin Care; Label; Phase; Biological; Chemicals; Lesion; Measurement; Oxidative Stress; Protocol; Protocols documentation; Source; cell type; System; genotoxicity; novel; Topical Drug Administration; administer topically; apply topically; deliver topically; topical administration; topical delivery; topical drug application; topical treatment; topically administered; topically applied; topically delivered; topically treated; treat topically; Topical application; Position; Positioning Attribute; Cytometry; Manufacturer; Manufacturer Name; preventing; prevent; Data; Clinical Evaluation; Clinical Testing; clinical test; research clinical testing; STTR; Small Business Technology Transfer Research; UVB; UVB radiation; Ultraviolet B Radiation; Validation; Exposure to ultraviolet radiation; UV exposure; Ultraviolet Radiation Related Exposure; Ultraviolet radiation exposure; ultraviolet exposure; UV Radiation Exposure; UV irradiated; UV irradiation; UV irridated; ultra violet irradiation; ultraviolet irradiation; UV damaged DNA; UV irradiated DNA; ultraviolet damaged DNA; ultraviolet induced DNA damage; ultraviolet irradiated DNA; UV induced DNA damage; designing; design; Population; Coupled; combinatorial; UV damage; ultra violet damage; ultraviolet damage; screening; Formulation; data pipeline

Skin is the first line of defense against the outside environment. When the skin barrier is breached by UV irradiation (UVR) or toxic chemicals, living cell layers including epidermal keratinocytes, melanocytes and dermal fibroblasts are subjected to DNA damage. If unrepaired, this damage can lead to photoaging and cutaneous carcinogenesis. In fact, skin cancer is by far the most common malignancy, accounting for more cases of cancer in the US and other countries than all other organs combined. Sunscreens have been used extensively with some success, but newer chemical filters suffer from serious shortcomings that make currently available products undesirable, and can be a risk to human health. Current strategies that examine the effectiveness and safety of organic UV filters need to be reassessed. These new strategies should test the biology of UVR in the presence of sunscreens. Further, they need to be high-throughput (HTP) in order to examine combinations of different chemical blockers that are present in most formulations, work in cultured cells for ease of screening, and be animal-friendly. In Phase I we showed proof-of-principle for a novel UVR HTP screening platform called "UValidate, employing mixed populations of isogenic keratinocytes and melanocytes derived from a single donor, exposed to two-compound combinations of sunscreen active ingredients in the presence and absence of short wave (UVB; 285-320), and long wave solar UVR (UVA1; 340-400 nm). These screens measure reactive oxygen species (ROS), DNA damage (CPDs), and micronuclei and show a complex interplay between solar UV wavelength, sunscreen blockers, and cell types. This highlights the pressing need for our platform, compared to currently-utilized assays that measure only the amount of UV that is blocked by spectrometry. In the proposed Phase II, we plan to test known chemical UV blockers for the cosmetic and skin-care markets as a contracting service, using a panel of donor-derived isogenic keratinocytes, melanocytes and fibroblast. At the completion of Phase II, if funded, UValidate will be offered as a service particularly suited for companies at the early stages of drug discovery, regulatory bodies and established pharma seeking to optimize formulation. Advances in this proposal include fluoro-tagging isogenic patient cell lines, establishing 2D and 3D cell cultures to determine individual responses to solar UV irradiation, utilization of reagents and assays to rapidly determine the types of DNA lesions and their repair, and design of AI software to more accurately determine damage. Control cells, generated using CRISPR technology, will reproduce DNA repair-compromised skin cells and 3D skin equivalents will also be included to ensure experimental rigor and reproducibility. The present crisis due to the lack of non- toxic sunscreens has led to class action lawsuits, banning of sunscreens in growing numbers worldwide, and recall of formulations from the market. Therefore, this technology is timely and needed with great expediency to address the urgent need for the discovery and testing of safe and effective UV blockers to prevent the most common, and often debilitating and deadly cancers that we face.

Public Health Relevance Statement:
Development of the UValidate platform for the chemical profiling of topically applied chemical agents. NARRATIVE Over-the-counter (OTC) active ingredients that block UVA and UVB are required in all sunscreens. Recently, the safety of these compounds has come under scrutiny both from a human health and ecological perspective. In Phase I, we successfully developed the UValidate platform, to address the immediate need to develop a better topical agent testing platform, particularly for UVA-blocking ingredients. UValidate is novel in that it can test the genotoxicity of sunscreens in conjunction with UVR treatment. In Phase II, we will develop UValidate from proof-of-principle, to a stable, integrated, HTP service platform able to measure biological endpoint directly relevant to sunscreen efficiency using cutting-edge donor-derived cellular models and complex combinational treatment regimes coupled with UVR exposure.

Project Terms:
Accounting; Aging; Algorithms; Animals; Biological Assay; Assay; Bioassay; Biologic Assays; Biology; Blood; Blood Reticuloendothelial System; Malignant Neoplasms; Cancers; Malignant Tumor; malignancy; neoplasm/cancer; Skin Cancer; Malignant Skin Neoplasm; malignant skin tumor; Cell Line; CellLine; Strains Cell Lines; cultured cell line; Cells; Cell Body; Cultured Cells; Complement; Complement Proteins; Contract Services; Cosmetics; cosmetic product; Diffusion; Digestion; DNA Damage; DNA Injury; DNA Repair; DNA Damage Repair; Unscheduled DNA Synthesis; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Environment; Face; faces; facial; Fibroblasts; Future; Health; Human; Modern Man; Industry; keratinocyte; Kinetics; Health Knowledge Attitudes Practice; Knowledge Attitudes Behaviors; Knowledge Attitudes Practice; Lead; Pb element; heavy metal Pb; heavy metal lead; melanocyte; Minerals; Mutagenicity Tests; Genetic Toxicity Tests; Genotoxicity Tests; Mutagen Screening; mutagen testing; Mutation; Genetic Alteration; Genetic Change; Genetic defect; genome mutation; Patients; Toxic Chemical; Toxic Substance; toxic compound; Poison; Protective Clothing; Quality Control; Reagent; Investigators; Researchers; Research Personnel; Risk; Safety; aged skin; skin photoaging; skin solar aging; Skin Aging; Cutaneous Disorder; Dermatoses; Skin Diseases; Skin Diseases and Manifestations; cutaneous disease; dermal disease; dermal disorder; skin disorder; Software; Computer software; Sunblock; Sunscreens; sun block; sun screen; Sunscreening Agents; Technology; Testing; Time; United States; Work; Zinc Oxide; Lassar's Paste; titanium dioxide; TiO2; titanium oxide; Measures; Skin Care; Short Waves; Reactive Oxygen Species; Active Oxygen; Oxygen Radicals; Pro-Oxidants; base; Organ; Label; improved; Surface; repaired; repair; Phase; Biological; biologic; Ensure; Chemicals; Lesion; Cutaneous; Dermal; Individual; Measurement; data retrieval; data storage; Data Storage and Retrieval; Funding; Tanning; Skin tanning; Exposure to; sun; The Sun; catalog; Catalogs; Spectrometry; Complex; cell type; 3-D; 3D; three dimensional; 3-Dimensional; Country; Services; Cancer Induction; carcinogenesis; success; basicity; alkalinity; novel; Topical Drug Administration; administer topically; apply topically; deliver topically; topical administration; topical delivery; topical drug application; topical treatment; topically administered; topically applied; topically delivered; topically treated; treat topically; Topical application; carcinogenicity; Modeling; High Throughput Assay; high throughput screening; drug discovery; Skin; Effectiveness; irradiation; preventing; prevent; Address; Reproducibility; Chemical Agents; research clinical testing; Clinical Evaluation; Clinical Testing; clinical test; Small Business Technology Transfer Research; STTR; Ultraviolet B Radiation; UVB; UVB radiation; ultra violet B radiation; Molecular; Development; developmental; Behavioral; Topical agent; ultraviolet irradiation; UV irradiated; UV irradiation; UV irridated; ultra violet irradiation; UVA induced; ultraviolet A induced; design; designing; Population; Coupled; data acquisition; combinatorial; Cell model; Cellular model; DNA lesion; FDA approved; cancer initiation; screening; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR; CRISPR/Cas system; sun damage; CRISPR/Cas technology; CRISPR approach; CRISPR based approach; CRISPR method; CRISPR methodology; CRISPR technique; CRISPR technology; CRISPR tools; CRISPR-CAS-9; CRISPR-based method; CRISPR-based technique; CRISPR-based technology; CRISPR-based tool; CRISPR/CAS approach; CRISPR/Cas method; CRISPR/Cas9; CRISPR/Cas9 technology; Cas nuclease technology; Clustered Regularly Interspaced Short Palindromic Repeats approach; Clustered Regularly Interspaced Short Palindromic Repeats method; Clustered Regularly Interspaced Short Palindromic Repeats methodology; Clustered Regularly Interspaced Short Palindromic Repeats technique; Clustered Regularly Interspaced Short Palindromic Repeats technology; skin barrier; cutaneous barrier; dermal barrier; epidermal barrier; Formulation; Knock-in; knockin; three dimensional cell culture; 3D cell culture; 3D culture; individual response; individualized response; deep learning; data pipeline; solar ultraviolet radiation; solar UV