Steve S. Lee, Ph.D. - US grants

My research examines childhood precursors of adolescent antisocial behavior (ASB) among separate samples of ADHD boys and girls. Unlike previous studies, key advantages here include: (a) rigorous diagnostic criteria for ADHD and comorbid conditions; (b) diverse predictors and outcomes; (c) sufficient ethnic and gender diversity; (d) a prospective design that enhances the specification of causal pathways. 177 boys (ages 6-12) participated in four separate naturalistic summer programs. Boys were observed during six-week camps with classroom and outdoor activities. About half of the boys had a diagnosis of ADHD and were actively treated with stimulants. The remaining boys served as normal comparisons. Observational data, including aggression and internalizing conditions were collected, as were peer sociometrics and academic achievement. Assessments were conducted prior to the camp and relevant measures were based on unmedicated periods. Five-years later, families were contacted for follow-up diagnostic interviews and questionnaires. Three samples have already been assessed and the final sample is currently being recruited. Similar summer programs were held for girls with and without ADHD (N=228); many fewer had medication histories. Four year follow-up visits will begin in Fall, 2001 and proceed annually until all three cohorts are assessed. The goal here is to (a) test childhood predictors of adolescent adjustment, with a focus on ASB and (b) test potential gender-specific predictions. Although possible cohort effects preclude precise comparisons, preliminary models will be constructed.

Project Summary This proposal aims to apply an imaging technique for three-dimensional (3D) visualization and mapping of the tumor and microenvironment including cancer cells, stromal cells, immune cells, and vascular cells. The capability to determine the location, density, and functional orientation of different cell populations throughout a tumor would be a powerful tool not only for cancer biology but also for improving diagnosis and advancing the development of effective therapies. Although immunohistochemistry (IHC)/immunofluorescence (IF) staining has long been used in cancer diagnostics, conventional IHC/IF methodology is based on localization of antigens in single thin sections. Considering that most cells in the field of view may be only fragments, interpretation of thin section staining is subject to multiple artifacts. Significantly, IHC/IF may fail to demonstrate important features such as pushing margin, lymphocytic infiltrate, or squamous change, leaving room for diagnostic and therapeutic uncertainties. Recent progress in tissue optical clearing and light microscopy enable 3D fluorescent imaging of normally opaque, thick tissues and organs. Dramatic examples include progress mapping neuronal connectivity in whole rodent brains at subcellular resolution. I sought to adapt these strategies to advance analysis of inflammatory infiltrates, drug delivery, and therapeutic responses in whole mouse tumors and human tumor biopsies. In order to advance 3D tumor imaging, I optimized optical clearing, cell staining, high resolution imaging, and computational reconstruction. Eventually, I developed a 3D tumor imaging method, Transparent Tumor Tomography (T3), as a tool to visualize and map tumors at single cell resolution. In this project, I will apply T3 for spatial analysis of tumor immunology and immunotherapy. T3-mediated 3D tumor imaging will provide integrated spatial information regarding antibody drug distribution and immune contexts in whole tumors which will be useful as a new assay tool for cancer immunotherapy research.