There are three aims to the proposed research, each with its own methodology but all with a common clinical objective - to identify and develop carbohydrate-specific tumor markers, with emphasis on hCG and the differential diagnosis of trophoblastic neoplasms. 1. Distribution of O-linked sugar units on pregnancy- and tumor-derived hCG. On hCG in pools of urine from normal pregnancy, 12-14% of the O-linked sugar units (4 total) are a hexasaccharide structure, NeuAc-Gal-GlcNAc- (NeuAc-Gal)-GalNAc-. In contrast, pilot studies show that the structure of those on hCG in the urines of choriocarcinoma subjects (2 examined) and in the media from choriocarcinoma cells were around 50% this hexasaccharide structure. The only published findings of this structure have been on cancer proteins. To confirm that hCG abundant in hexasaccharides is a choriocarcinoma marker, the structures of O-linked sugars on hCG molecules in the urines of 12-15 additional subjects will be examined (methods: immunoprecipitation of hCG, beta-elimination/labeling of O-linked sugar units, chromatographic comparison with oligosaccharide standards). To determine if the abundance of hexasaccharide is correlated with the malignance of trophoblastic disease, that on urinary hCG molecules from subjects with the more benign invasive/partial/complete mole will also be examined. O-linked sugar units on hCG from nontrophoblastic neoplasms, and on other urinary and tissue glycoproteins will be examined. 2. Development on trophoblast disease-specific assays. Although trophoblast disease is rare in the U.S.A. (1 in 1500 pregnancies), a 10 X higher occurrence is found in several African and Asian nations. To generate a disease-specific assay, the beta subunit C-terminal peptide containing the hexasaccharides on choriocarcinoma hCG will be isolated and utilized as immunogen for generating sheep polyclonal antisera, for monoclonal antibody approaches, and as domain for a lectin-antibody system. 3. Glycosyltransferase activities in normal, neoplastic, and induced or transformed cells. To investigate the pathways or mechanisms by which changes in protein glycosylation accompany cancer, glycosyltransferase activities will be examined in trophoblast explant cultures. Effects of virus and chemical transformation, growth factors, and other inducers/suppressants of cellular differentiation on enzyme activities will be examined; where appropriate kinetic aspects of activity changes and effects of DNA, RNA and protein synthesis inhibitors will be investigated.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA044131-01
Application #
3186694
Study Section
Pathology B Study Section (PTHB)
Project Start
1986-05-01
Project End
1989-04-30
Budget Start
1986-05-01
Budget End
1987-04-30
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Iles, Ray K; Cole, Laurence A; Butler, Stephen A (2014) Direct analysis of hCG?cf glycosylation in normal and aberrant pregnancy by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Int J Mol Sci 15:10067-82
Elliott, M M; Kardana, A; Lustbader, J W et al. (1997) Carbohydrate and peptide structure of the alpha- and beta-subunits of human chorionic gonadotropin from normal and aberrant pregnancy and choriocarcinoma. Endocrine 7:15-32
Kardana, A; Cole, L A (1997) The stability of hCG and free beta-subunit in serum samples. Prenat Diagn 17:141-7
Cole, L A; Kohorn, E I; Kim, G S (1994) Detecting and monitoring trophoblastic disease. New perspectives on measuring human chorionic gonadotropin levels. J Reprod Med 39:193-200
Cole, L A; Kardana, A; Seifer, D B et al. (1994) Urine hCG beta-subunit core fragment, a sensitive test for ectopic pregnancy. J Clin Endocrinol Metab 78:497-9
Cole, L A; Seifer, D B; Kardana, A et al. (1993) Selecting human chorionic gonadotropin immunoassays: consideration of cross-reacting molecules in first-trimester pregnancy serum and urine. Am J Obstet Gynecol 168:1580-6
Cole, L A; Kardana, A; Park, S Y et al. (1993) The deactivation of hCG by nicking and dissociation. J Clin Endocrinol Metab 76:704-10
Cole, L A; Kardana, A (1992) Discordant results in human chorionic gonadotropin assays. Clin Chem 38:263-70
Kardana, A; Cole, L A (1992) Polypeptide nicks cause erroneous results in assays of human chorionic gonadotropin free beta-subunit. Clin Chem 38:26-33
Cole, L A; Kardana, A; Andrade-Gordon, P et al. (1991) The heterogeneity of human chorionic gonadotropin (hCG). III. The occurrence and biological and immunological activities of nicked hCG. Endocrinology 129:1559-67

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