The long range objective of this proposal is to elucidate the importance of two classes of mechanisms of deregulation of proto- oncogenes in human tumors; that is, mutations in the proto- oncogenes itself (cis-acting) and mutations in trans-acting regulators. The studies focus on c-myc because this gene is expressed in many different types of human tumors. What is the importance of c-myc expression in these tumors? The expression is presumed to be due to mutation that affect the myc regulatory circuit. What are the different types of mutations that have this effect? The aims of this proposal include an analysis of cis- acting alterations in two different myc loci clones from human tumors. In one case, a LINE-1 sequence (one of a family of repetitive transposable elements present in thousands of copies in the human genome) has become inserted into intron 2 of myc. The insertion is tumor specific. The insertion will be cloned and sequenced and analyzed for the presence of enhancer elements and for the capacity to code for proteins involved in the process of transposition. The second locus under study produces myc message levels 50 times greater than those found in normal leukocytes. The locus contains multiple deletions, insertions, duplications, and point mutations. The exact role of each of these aberrations in the deregulation will e determine.
A second aim of this proposal is to analyze trans-acting alterations that result in deregulation of c-myc in two thirds of human adenocarcinomas of the colon. Experiments employing somatic cell hybrids between cells expressing regulated c-myc and colon carcinoma cells expressing deregulated myc result in hybrid cells in which myc is regulated normally. A working hypothesis, based on the results of this experiments, is that genetic defects in the colon carcimona cell parent have inactivated both alleles of a trans-acting locus that regulates myc (such as a repressor) and that the defects are complemented by an intact locus in the parent expressing regulated myc. These experiments will be pursued to determine how many such trans-acting loci there might be, what human chromosomes they map to, an what the molecular mechanism is by which lesions in these loci result in deregulation of c-myc. This work might ultimately have clinical significance in that the normal alleles of these genes could be cloned and supplied to the tumor cells through the use of suitable vectors such aS retroviruses. Through the use of such """"""""gene transplant therapy"""""""" the transformed phenotype of the tumor cells might be reversed.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA040636-07
Application #
3180923
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1985-07-01
Project End
1994-01-31
Budget Start
1992-04-17
Budget End
1993-01-31
Support Year
7
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Institute for Cancer Research
Department
Type
DUNS #
872612445
City
Philadelphia
State
PA
Country
United States
Zip Code
19111
Laurence, J; Grimison, B; Rodriguez-Alfageme, C et al. (1993) A model system for regulation of chronic HIV-1 infection in peripheral B lymphocytes. Virology 196:433-41
Astrin, S M; Schattner, E; Laurence, J et al. (1992) Does HIV infection of B lymphocytes initiate AIDS lymphoma? Detection by PCR of viral sequences in lymphoma tissue. Curr Top Microbiol Immunol 182:399-407
Rodriguez-Alfageme, C; Stanbridge, E J; Astrin, S M (1992) Suppression of deregulated c-MYC expression in human colon carcinoma cells by chromosome 5 transfer. Proc Natl Acad Sci U S A 89:1482-6
Laurence, J; Astrin, S M (1991) Human immunodeficiency virus induction of malignant transformation in human B lymphocytes. Proc Natl Acad Sci U S A 88:7635-9
Morse, B; South, V J; Rothberg, P G et al. (1989) Somatic mutation and transcriptional deregulation of myc in endemic Burkitt's lymphoma disease: heptamer-nonamer recognition mistakes? Mol Cell Biol 9:74-82
Erisman, M D; Scott, J K; Astrin, S M (1989) Evidence that the familial adenomatous polyposis gene is involved in a subset of colon cancers with a complementable defect in c-myc regulation. Proc Natl Acad Sci U S A 86:4264-8
Astrin, S M; Costanzi, C (1989) The molecular genetics of colon cancer. Semin Oncol 16:138-47
Erisman, M D; Litwin, S; Keidan, R D et al. (1988) Noncorrelation of the expression of the c-myc oncogene in colorectal carcinoma with recurrence of disease or patient survival. Cancer Res 48:1350-5
Erisman, M D; Scott, J K; Watt, R A et al. (1988) The c-myc protein is constitutively expressed at elevated levels in colorectal carcinoma cell lines. Oncogene 2:367-78