Translational Investigation for Tea Attenuate Smoking-Induced Disease---Breast Cancer and Atherosclerosis as a Mechanistic Study

Our previous research has demonstrated that the a9-nicotinic acetylcholine receptor (a9-nAchR) can be detected at high levels in human breast cancer tissue (>8-fold increase, JNCI 2010; 102(17):1322-1335). The JNCI editor Professor R. Ilona Linnoila wrote a special commentary indicating that a9-nAchR-mediated carcinogenic signals play a major clinical role in breast tumors (JNCI 2010; 102(17):1298-1299). We have previously proposed that blocking a9-nAchR in breast cancer cells could be the best strategy for preventing the deleterious effects induced by smoking (Clinical Cancer Research 2011, 17:3533-3541, review article). The current study proposes that compounds found in tea could have preventive effects on smoking-induced disease and that the mechanisms of these potential effects should be investigated by clinical trial translational research methods. 1. Tea capsule preparation and analysis: 1-1 Tea components analysis: The most popular Taiwanese teas will be used, including Oolong, Pu-erh, Green, Black, Bao-Chung, Jasmine, etc. 1-2 Tea extract preparations: Tea components’ abilities to attenuate nicotine-induced effects in vitro and in vivo will be studied. 1-3 Preparation of GMP-level tea capsules will be used for clinical trials. 2. Clinical trials for tea components that attenuate smoking-induced deleterious effects: 2-1 Classification of test volunteers, blood and urine sample collection for routine analysis. 2-2 Human leukocytic bio-bank: Analysis of the nAChR expression levels in mononuclear cells before, during and after tea capsule treatment by flow cytometry analysis. 2-3 Molecular markers identification in early-onset, triple-negative and/or smoker breast cancer populations: miRNA signature profiles will be detected in the blood serum by ChiP array in breast cancer patients before and after surgery. 2-4 The identified miRNAs will be used as molecular markers to evaluate the ability of tea capsules to attenuate smoking-induced deleterious effects in the clinical trial patients. 2-5 Tea components as chemo- or radio-therapeutic adjuvants will be evaluated based on alterations in the specific miRNAs that will be used as molecular markers. 3. In vitro cell culture research models: 3-1 Human leukocyte bio-bank establishment: Determination of the a9-nAChR expression levels in leukocytes isolated from clinical trial patients by flow cytometry methods. 3-2 Purification of mononuclear cells from test individuals by magnetic-antibody harvest methods: Primary culture to evaluate nicotine-induced effects. 3-3 Evaluation of the ability of tea capsules to attenuate nicotine-induced deleterious effects. 3-4 Arthrosclerosis-related test models: Foam-cell formation, LDL expression in monocytes, interactions between monocytic and endothelial cells, etc. 3-5 Anti-tumor models in vitro: Human endothelial cell culture for angiogenesis formation, breast cancer cell invasion assays, migration assays and interaction of nAcHR and cell membrane proteins by FRET and FLIM assays. 3-6 Development of a disease-specific miRNA profile as a diagnosis chip array. 4. In vivo animal research models: (please refer to the proposal for a detailed description) 4-1 Tea components inhibit nicotine-induced arthrosclerosis model. 4-2 Tea components inhibit nicotine-induced neovascular angiogenesis model. 4-3 Tea components inhibit tumor metastasis model. 4-4 Tea components as chemo- or radio-therapeutic adjuvants, personalized anti-tumor model.