The long term goal of this project is to provide the applicant with the proper training in molecular genetics and cell biology to become an independent investigator in the field of atherosclerosis. To accomplish this goal, a training program has been designed which consists of a well balanced didactic component coupled with closely supervised laboratory training in molecular and cellular biology. The goal of the laboratory work will be to use a genetic, molecular and cellular approach to better understand the function of endothelial adhesion molecules, in particular intercellular adhesion molecule-1 (ICAM-1). The interaction of endothelial adhesion molecules with circulating elements in the blood is felt to play a crucial role in inflammation, platelet adhesion, and thrombosis. Creation of a mouse mutant deficient in ICAM-1 by homologous recombination in embryonic stem cells and generation of transgenic mice in which ICAM-1 is expressed at an increased level in either a normal or aberrant pattern would help to define the significance of ICAM-1 in vivo. The following specific research aims are proposed: (1) clone and sequence the full length mouse cDNA for ICAM-1; (2) express the mouse cDNA as a fusion protein and use this to generate antibodies; (3) clone and sequence relevant portions of the mouse gene; (4) construct transgenic mice in which ICAM-1 is expressed at an increased level in either a normal or aberrant pattern; (5) production of a mouse mutant deficient in ICAM-1 by homologous recombination in embryonic stem cells. Animal models with a receptor deficiency or excess have prove invaluable in understanding the function of other surface receptors such as the LDL receptor and would provide an important model in which to study the function of ICAM-1 in vivo. The laboratory projects as outlined in conjunction with a didactic program consisting of Ph.D. level course work, laboratory meetings, literature seminars, and departmental seminars with supervision by an academic committee should provide the applicant with the experience and knowledge needed to embark on a career studying the cellular and molecular genetic aspects of endothelial and vascular biology as they relate to atherosclerosis.
