Despite many advances in the treatment of cancer in recent years, most malignant tumors are currently incurable unless they are detected at an early stage. One approach to increasing the success of cancer therapy in this setting is to augment the ability of the body's immune defense system to destroy residual cancer cells after surgery, radiation or chemotherapy. A major component of the body's immune defense system is the major histocompatibility (MHC) Class I gene system also known as the human leukocyte antigen (HLA) Class I gene complex. Expression of the genes of the HLA Class I complex plays a critical role in the function of cell-mediated killing of tumor cells. Another approach which has allowed cure of previously incurable patients with hematologic malignancies is bone marrow transplantation. Tbe ability to utilize this treatment modality is also dependent on the type of HLA Class I gene expression in the bone marrow donor and recipient. The long term objectives of this project are to elucidate cell-type specific and allele-specific mechanisms responsible for the constitutive and immune interferon (IFNgamma) mediated regulation of HLA Class I gene expression in human hematopoietic tumor cells and lymphoid hematopoietic cells. The ultimate goal of these studies is to provide a rational basis for therapeutic manipulation of HLA Class I gene expression to facilitate host defense killing of tumor cells and to facilitate the ability to carry out bone marrow transplantation across MHC Class I barriers. Several specific questions will be addressed to achieve the long term objectives of this project: 1) What is the nature and specificity of a newly described constitutive control element in the 5' flanking region of the HLA-A2 I gene and the trans-acting nuclear protein that binds to this element? 2) What are the exact cis-acting sequences of a novel 3'interferon response element and corresponding nuclear protein binding complex present in the HLA-A2 gene, and is this element allele specific? These questions will be answered by functional analysis of the cis-acting regulatory sequences through in vitro mutagenesis and DNA-mediated gene transfer into human tumor and lymphoid cells. The trans-acting factors that mediate the effects of both the constitutive and interferon responsive regulatory sequences will be characterized by employing a combination of biochemical purification procedures as well as direct screening of cDNA expression libraries to obtain purified factors and the genes that encode them. These factors will then be analyzed functionally in biochemically defined in vitro transcription assays and by surrogate genetic techniques. The interactions between the constitutive and interferon responsive control elements of HLA Class I gene expression will be tested in DNA-mediated gene transfer genetic assays and in vitro biochemical assays. Besides the specific information related to the regulation of HLA Class I gene regulation in human tumor and lymphoid cells, which ultimately could have relatively direct clinical applications, it is anticipated that knowledge gained about the basic mechanisms controlling Class I gene expression could have broad application in understanding and manipulating this critical component of the human immune system.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA045634-08
Application #
2091945
Study Section
Pathology B Study Section (PTHB)
Project Start
1990-09-20
Project End
1996-06-30
Budget Start
1994-07-01
Budget End
1995-06-30
Support Year
8
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Snyder, S R; Wang, J; Waring, J F et al. (2001) Identification of CCAAT displacement protein (CDP/cut) as a locus-specific repressor of major histocompatibility complex gene expression in human tumor cells. J Biol Chem 276:5323-30
Snyder, S R; Waring, J F; Zhu, S Z et al. (2001) A 3'-transcribed region of the HLA-A2 gene mediates posttranscriptional stimulation by IFN-gamma. J Immunol 166:3966-74
Gustafson, K S; Ginder, G D (1996) Interferon-gamma induction of the human leukocyte antigen-E gene is mediated through binding of a complex containing STAT1alpha to a distinct interferon-gamma-responsive element. J Biol Chem 271:20035-46
Waring, J F; Radford, J E; Burns, L J et al. (1995) The human leukocyte antigen A2 interferon-stimulated response element consensus sequence binds a nuclear factor required for constitutive expression. J Biol Chem 270:12276-85
Burns, L J; Waring, J F; Reuter, J J et al. (1993) Only the HLA class I gene minimal promoter elements are required for transactivation by human cytomegalovirus immediate early genes. Blood 81:1558-66
Radford Jr, J E; Waring, J F; Pohlman, J K et al. (1993) Stimulation of MHC class I transcription by interferon-gamma involves a non-A, non-C kinase in addition to protein kinase C. J Interferon Res 13:133-41
Radford Jr, J E; Chen, E; Hromas, R et al. (1991) Cell-type specificity of interferon-gamma-mediated HLA class I gene transcription in human hematopoietic tumor cells. Blood 77:2008-15
Chen, E; Karr, R W; Ginder, G D (1987) Negative and positive regulation of human leukocyte antigen class I gene transcription in K562 leukemia cells. Mol Cell Biol 7:4572-5