Date Completed: 2/8/08
Faculty Name: Laura Timares
UAB Department: Dermatology, Division of Human Gene Therapy
UAB School: School of Medicine
Campus Address: BMR2-542
Campus Zip: 2172
Email Address: firstname.lastname@example.org
Research Program Area: Tumor Immunology
Project Title: Topical immunization to protect against skin cancer development
Project Status: Already up-and-running
Proposed Start Date: April 14
Proposed End Date: July 25
Number of Weeks of Internship: 15
Number of Interns: 1
Other faculty, staff, or graduate students who may help supervise intern: Hee Kyung Kim
Expected Number of Work Hours Per Week: 37.5 hours
Expected Work Schedule for Intern: Flexible, intern can largely set his or her own schedule
Category of Research: Animal Research
Cancer Research: Skin; Chemical Carcinogens
Project Description: Squamous cell carcinoma is a common skin cancer that arises due to exposure to environmental (UV) or chemical carcinogens. Skin tumors generated by chemical carcinogens are strongly associated with a specific point mutation in the Ras proto-oncogene, an important regulatory GTPase, that promotes uncontrollable cell growth and proliferation.
We hypothesize that if we could generate a cellular immune response against the mutant Ras epitope (*Ras), we could immunze against the development of skin cancer. However, because the immunogenicity of a single point mutation of a self protein is low, new immunization strategies must be developed to enhance the immune recognition of the *Ras point mutation. Dendritic cells are potent antigen presenting cells that may be manipulated to focus immune recognition of the *Ras epitope. We have generated a genetic immunization vaccine that encodes a fusion protein of the *Ras epitope to GFP linked to ubiquitin (Ub-*Ras-GFP) and promotes loading onto MHC I for presentation to CD8 T cells. We have also shown that topical vaccination with this DNA-based vaccine can induce a cellular response to *Ras peptide challenge that is superior to standard methods of vaccination with *Ras peptide. The CaRES student may be involved in one of the following projects:
1) Assess the adjuvant (enhancing) effect of Toll-receptor ligand using in conjunction with topical vaccination on the generation of *Ras-specific CD8+ cytotoxic T lymphocyte (CTL) activity. Assessed by performing in vivo CTL assays using flow cytometric analysis.
2) Generate lentiviral vectors encoding Ub-*Ras-GFP and parallel control constructs. Infect skin-derived dendritic cells (DCs) to use as vaccinating agents. Develop stable DC clones.
3) Test the prophylactic protection against chemically (DMBA) induced *Ras expressing tumors on mice. Conduct chemical carcinogenesis protocols, and monitor the numbers and growth kinetics of tumors that grow in vaccinated and unvaccinated mice.
Intern's Anticipated Duties:
Duty 1: Working with animals, immunologic assays, plasmid purification, use of chemical carcinogen DMBA
Duty 2: Molecular biology - lentivirus generation, tissue culture, infection procedures, flow cytometry
Duty 3: Keeping a complete lab notebook and write-up of the project. Must complete certification for completing online courses on animal handling and lab safety
Preceptor will provide intern with access to the following: office or desk space; computer and printer; laboratory work bench space; equipment needed to complete project; supplies needed to complete project
Likelihood of authoring publications: Very likely
Background, education, experience, or expertise preferred: Animal Research; Cell Biology; Environmental Carcinogenesis; Basic Knowledge of Lab Skills; Advanced Knowledge of Lab Skills; Literature Review Skills; Molecular Biology; Immunology
This faculty member has been a CaRES Preceptor for three or more summers.
Intern 1: Kyle Rudemiller