The major goals of this research are to use genetic approaches to determine: (a) whether metallothioneins (MT) are essential during mammalian development and physiology, (b) whether different isoforms of MT have distinct functions and (c) whether MTs can play a role in retarding malignant transformation. The MT-I and MT-II genes are expressed in most organs, they are inducible by a variety of metals and hormones, and they probably have a similar, general function. These MTs will be disrupted by homologous recombination in embryonic stem cells and those cells will be used to generate mice lacking MT-I, MT-II or both isoforms. The effect of these mutations on normal development and physiological responses will be examined. Mice over-expressing MT-I or MT-III (an unusual MT normally expressed only in the brain) have been generated by microinjection of marked versions of these genes flanked by presumptive locus control regions from the MT locus. These transgenic mice will be used to determine if excess or ectopic expression of MT affects development or physiology. Mice expressing reduced or elevated levels of MT will then be crossed with two different transgenic lines of mice with predispositions to develop hepatocellular carcinoma. If MT plays a role in retarding tumorigenesis, then we predict that hepatic tumors will develop more slowly in mice expressing excess MT and more rapidly in mice expressing reduced levels of MT.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA061268-04
Application #
2008334
Study Section
Special Emphasis Panel (SRC (54))
Program Officer
Liu, Yung-Pin
Project Start
1994-02-01
Project End
1998-12-31
Budget Start
1997-01-01
Budget End
1998-12-31
Support Year
4
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Quaife, C J; Cherne, R L; Newcomb, T G et al. (1999) Metallothionein overexpression suppresses hepatic hyperplasia induced by hepatitis B surface antigen. Toxicol Appl Pharmacol 155:107-16
Quaife, C J; Kelly, E J; Masters, B A et al. (1998) Ectopic expression of metallothionein-III causes pancreatic acinar cell necrosis in transgenic mice. Toxicol Appl Pharmacol 148:148-57
Erickson, J C; Hollopeter, G; Thomas, S A et al. (1997) Disruption of the metallothionein-III gene in mice: analysis of brain zinc, behavior, and neuron vulnerability to metals, aging, and seizures. J Neurosci 17:1271-81
Kelly, E J; Sandgren, E P; Brinster, R L et al. (1997) A pair of adjacent glucocorticoid response elements regulate expression of two mouse metallothionein genes. Proc Natl Acad Sci U S A 94:10045-50
Palmiter, R D; Cole, T B; Findley, S D (1996) ZnT-2, a mammalian protein that confers resistance to zinc by facilitating vesicular sequestration. EMBO J 15:1784-91
Dalton, T; Fu, K; Palmiter, R D et al. (1996) Transgenic mice that overexpress metallothionein-I resist dietary zinc deficiency. J Nutr 126:825-33
Lee, D K; Fu, K; Liang, L et al. (1996) Transgenic mouse blastocysts that overexpress metallothionein-I resist cadmium toxicity in vitro. Mol Reprod Dev 43:158-66
Liu, Y P; Liu, J; Palmiter, R D et al. (1996) Metallothionein-I-transgenic mice are not protected from acute cadmium-metallothionein-induced nephrotoxicity. Toxicol Appl Pharmacol 137:307-15
Kelly, E J; Palmiter, R D (1996) A murine model of Menkes disease reveals a physiological function of metallothionein. Nat Genet 13:219-22
Kelly, E J; Quaife, C J; Froelick, G J et al. (1996) Metallothionein I and II protect against zinc deficiency and zinc toxicity in mice. J Nutr 126:1782-90

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