Our hypothesis is that nicotine and/or traditional tobacco smoke exposure will modify RNA biology via inducing alternative AP patterns and isomiR formation. To investigate these two hypotheses, we are now conducting preliminary pilot studies using normal and cancer cell lines, with and without nicotine and tobacco smoke condensate exposure. We are conducting a novel 3 prime UTR-specific RNA-sequencing method to examine APA patterns. Depending on the results of these analyses, we will subsequently conduct an analysis of human lung cancer and normal tissue from the NCI-MD case control study to ask this question in human lung cancer. To investigate the second part of this hypothesis, we are conducting a similar protocol but instead we are subjecting the samples to miRNA sequencing. Expression differences in isomiR formation will be further characterized for functional significant in follow up studies.