We will continue our studies of the human apolipoprotein genes to determine what role genetic variation in them might play in atherosclerosis susceptibility. Our major focus will be on the apo A-I and apo E genes. There are two specific aims: 1) Transcriptional regulation of apolipoprotein gene expression: The cis-Acting DNA elements that control transcription of the apo A-I, E and CIII genes will be defined. Each of the cloned genes will be manipulated in vitro and then transfected into differentiated human cell cultures. Transient expression will be assessed by either RNA or protein analysis. Transcriptionally active DNA elements will be defined by deletion analysis and by site specific mutagenesis. Initially, apolipoprotein gene expressing HepG2 and non-expressing Hela cells will be used, followed by studies in macrophage and intestinal cell lines. Trans-acting protein factors that bind to these regions will be characterized and purified using mobility shift and in vitro transcription assays. Finally, the transgenic mouse model will be used to verify in vivo the DNA sequence requirements for tissue specific expression. In this model we will also test whether apo A-I gene transcriptional regulation affects plasma HDL levels. 2) Clinically significant apolipoprotein genetic variation. RFLP's at the chromosome 11 (apo A-I) and chromosome 19 (apo E) gene loci will be assessed for association with myocardial infarction or its associated lipid, lipoprotein or apolipoprotein abnormalities. In addition, these RFLP's will be used in studying families with dyslipoproteinemia phenotypes to see if either of these loci are linked to particular clinical disorders. Partial or complete genomic libraries in plasmid or lambda vectors will be constructed from individuals who may have as their underlying defect a mutation in the apo A-I or apo E genes to isolate the gene harboring the suspected mutation. Structural and functional studies will be conducted to determine the exact nature of the mutation. It will then be possible to probe for the specific gene defect in the general population or in a subset with a particular clinical phenotype to determine the frequency of the mutation. This information could form the basis of new tests for atherosclerosis susceptibility in presymptomatic individuals.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL033714-07
Application #
3345840
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1984-07-01
Project End
1992-06-30
Budget Start
1990-07-01
Budget End
1991-06-30
Support Year
7
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Rockefeller University
Department
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Liu, Simin; Song, Yiqing; Hu, Frank B et al. (2004) A prospective study of the APOA1 XmnI and APOC3 SstI polymorphisms in the APOA1/C3/A4 gene cluster and risk of incident myocardial infarction in men. Atherosclerosis 177:119-26
Liu, Simin; Ma, Jing; Ridker, Paul M et al. (2003) A prospective study of the association between APOE genotype and the risk of myocardial infarction among apparently healthy men. Atherosclerosis 166:323-9
Trogan, Eugene; Choudhury, Robin P; Dansky, Hayes M et al. (2002) Laser capture microdissection analysis of gene expression in macrophages from atherosclerotic lesions of apolipoprotein E-deficient mice. Proc Natl Acad Sci U S A 99:2234-9
Han, Zhihua; Heath, Simon C; Shmulewitz, Dvora et al. (2002) Candidate genes involved in cardiovascular risk factors by a family-based association study on the island of Kosrae, Federated States of Micronesia. Am J Med Genet 110:234-42
Rong, J X; Li, J; Reis, E D et al. (2001) Elevating high-density lipoprotein cholesterol in apolipoprotein E-deficient mice remodels advanced atherosclerotic lesions by decreasing macrophage and increasing smooth muscle cell content. Circulation 104:2447-52
Tamminen, M; Mottino, G; Qiao, J H et al. (1999) Ultrastructure of early lipid accumulation in ApoE-deficient mice. Arterioscler Thromb Vasc Biol 19:847-53
Dansky, H M; Charlton, S A; Barlow, C B et al. (1999) Apo A-I inhibits foam cell formation in Apo E-deficient mice after monocyte adherence to endothelium. J Clin Invest 104:31-9
Weng, W; Brandenburg, N A; Zhong, S et al. (1999) ApoA-II maintains HDL levels in part by inhibition of hepatic lipase. Studies In apoA-II and hepatic lipase double knockout mice. J Lipid Res 40:1064-70
Zaiou, M; Azrolan, N; Hayek, T et al. (1998) The full induction of human apoprotein A-I gene expression by the experimental nephrotic syndrome in transgenic mice depends on cis-acting elements in the proximal 256 base-pair promoter region and the trans-acting factor early growth response factor 1. J Clin Invest 101:1699-707
Weng, W; Breslow, J L (1996) Dramatically decreased high density lipoprotein cholesterol, increased remnant clearance, and insulin hypersensitivity in apolipoprotein A-II knockout mice suggest a complex role for apolipoprotein A-II in atherosclerosis susceptibility. Proc Natl Acad Sci U S A 93:14788-94

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