When human and fungal worlds collide - a systems biology approach to understanding the dynamic interactions between human bronchial epithelial cells and conidiospores of Aspergillus fumigatus.

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My research program will improve the scientific understanding, at the molecular and cellular levels, of how organisms from two disparate kingdoms (Animalia and Fungi) dynamically interact with one another, during co-culture. Specifically, I have selected human bronchial airway epithelial cells and spores from the fungus, Aspergillus fumigatus, due to the ubiquity of A. fumigatus whose spores are inhaled by all of us, each and every day. Furthermore, the genomes of both organisms have been sequenced and are well annotated, which will allow good biological inferences and hypothesis testing based on the gene expression (RNA) datasets generated through my shorter-term objectives (detailed below). Over the medium to longer term, I aim to extend my NSERC research program to incorporate proteomic and metabolomic studies, as well as direct experimental manipulations (eg, site-directed mutagenesis or siRNA knock-down) of genes that my current short-term objectives identify as important players in dynamic interactions between these spores and mammalian cells. Future refinement of the co-culture models will also be important, in order to explore additional cell types in the airway (eg, alveolar cells and dendritic cells).***My working hypothesis is that the interaction of A. fumigatus conidiospores with human primary bronchial epithelial cells leads to dynamic changes in the molecular profiles (transcriptome, proteome, metabolome) of both the fungal and human organisms.****Short-term objectives over the next 5 years of grant funding:***1. Develop an interaction model of A. fumigatus conidia with human primary bronchial epithelial cells, and measure spore uptake, as well as spore and cell death, over time using a variety of methods including fluorescence microscopy.***2. Use both unbiased and targeted molecular profiling technology (eg, RNA-Seq, nanoString nCounterr, SmartFlareT) to simultaneously measure dual organism gene expression changes in primary bronchial epithelial cells and A. fumigatus spores, during co-culture interaction.***3. Further develop and test a novel dual organism microfluidic-based device that will allow single human airway cells to come into contact with and interact with single A. fumigatus conidia.***The significance of the work outlined in this research program is several-fold: it will bring a new and unbiased transcriptomic perspective to the investigation of the interaction of A. fumigatus conidiospores with bronchial epithelial cells; application of nascent technologies including single cell/SmartFlare analysis and microfluidics will be applicable to other areas of research related to how organisms from different species interact at the cellular and molecular level; and multidisciplinary training of graduate and undergraduate students in areas as diverse as engineering, molecular biology, bioinformatics and biostatistics, and fungal and human cell biology.**