P53 Mutation Screening by Multiplex Sequencing

We will develop a rapid, cost-effective screening method for detecting all mutations in the p53gene. The goal of Phase I is to optimize conditions for direct genomic sequencing of PCR products fromhuman tumor cell lines. Both malignant mesotheliomas and sarcomas have been shown to be inducedby different chemical carcinogens. Forty malignant mesothelioma and sarcoma cell lines, for whichmutations have previously been detected, will be studied. These samples will allow the determinationof the efficiency and applicability of multiplex sequencing for scanning genomic DNA for point mutations.Phase II will involve the study of large numbers of malignant mesotheliomas. These experiments willdetermine if specific codons or exons in p53 are mutated in malignant mesotheliomas and if there is anycorrelation between the site of mutation and the clinical outcome. Overall, this research willdemonstrate the feasibility of using a multiplex sequencing approach for mutation scanning in humanclinical samples. While the specific aims of this project are to determine the location of p53 mutationsin malignant mesotheliomas, the methods developed will allow one to screen other forms of cancer formutations in p53 as well as in other genes.