title: Using HSV-thymidine kinase for safety in an allogeneic salivary graft cell line.
abstract: Extreme salivary hypofunction is a result of tissue damage caused by irradiation therapy for cancer in the head and neck region. Unfortunately, there is no currently satisfactory treatment for this condition that affects up to 40,000 people in the United States every year. As a novel approach to managing this problem, we are attempting to develop an orally implantable, fluid-secreting device (an artificial salivary gland). We are using the well-studied HSG salivary cell line as a potential allogeneic graft cell for this device. One drawback of using a cell line is the potential for malignant transformation. If such an untoward response occurred, the device could be removed. However, in the event that any HSG cells escaped, we wished to provide additional patient protection. Accordingly, we have engineered HSG cells with a hybrid adeno-retroviral vector, AdLTR.CMV-tk, to express the herpes simplex virus thymidine kinase (HSV-tk) suicide gene as a novel safety factor. Cells were grown on plastic plates or on poly-L-lactic acid disks and then transduced with different multiplicities of infection (MOIs) of the hybrid vector. Thereafter, various concentrations of ganciclovir (GCV) were added, and cell viability was tested. Transduced HSG cells expressed HSV-tk and were sensitive to GCV treatment. Maximal effects were seen at a MOI of 10 with 50 microM of GCV, achieving 95% cell killing on the poly-L-lactic acid substrate. These results suggest that engineering the expression of a suicide gene in an allogeneic graft cell may provide additional safety for use in an artificial salivary gland device.
authors.affiliation: Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Aframian
authors.forename: C
authors.initials: C
authors.lastname: Zheng
authors.forename: C M
authors.initials: CM
authors.lastname: Goldsmith
authors.forename: J
authors.initials: J
authors.lastname: Nikolovski
authors.forename: E
authors.initials: E
authors.lastname: Cukierman
authors.forename: K M
authors.initials: KM
authors.lastname: Yamada
authors.forename: D J
authors.initials: DJ
authors.lastname: Mooney
authors.forename: H
authors.initials: H
authors.lastname: Birkedal-Hansen
authors.forename: B J
authors.initials: BJ
authors.lastname: Baum
mesh_terms: D001187:Artificial Organs; D001705:Bioprosthesis; D002460:Cell Line; D005822:Genetic Vectors; D006801:Humans; D012469:Salivary Glands / Q000637:transplantation*; D018139:Simplexvirus; D013937:Thymidine Kinase / Q000235:genetics; D023822:Tissue Engineering; D014184:Transplantation, Homologous
chemical_list: D013937:Thymidine Kinase