The long-term goal of this project is to better understand how cells communicate with each other. This project focuses on the relatively unexplored process of cell-cell communication via microRNAs. A major roadblock to understanding microRNA-mediated cell-cell communication has been the inability to distinguish microRNAs produced by donor and recipient cells. This project will employ an innovative approach to get around this roadblock. The successful completion of this work will lead to a better understanding of microRNA-mediated cell-cell communication and a new set of resources and tools for investigating cell-cell interactions in a variety of organisms, including humans.
MicroRNAs (miRNAs) are small non-coding RNAs with significant regulatory roles in all physiological processes. miRNAs were thought to be stable only inside cells and rapidly degraded by enzymes when outside the cell. Recent work has demonstrated that this is not the case: cells can export miRNAs through multiple mechanisms, affecting other cells either next door or at great distances away. Past work has typically focused on one miRNA at a time because it is impossible to distinguish miRNA molecules made by one cell from those made by another. Major questions remain unanswered: Which miRNAs are transferred? What mechanisms do cells use to transfer them? To overcome hurdles and answer these questions, expertise in miRNA biology, synthetic biology and genome editing will be leveraged. Two features of non-mammalian systems will be adopted: a protozoan enzyme to label RNA, and CRISPR/Cas to engineer custom genetic circuits and introduce genome modifications. Combining cells engineered to label RNA with those that do not label RNA will enable the identification of miRNA species that are transferred between cells. Implementation and stable integration of custom synthetic biology circuits in human cells will permit the reliable monitoring of miRNA levels. Selective pharmacological blocking of inter-cellular communication mechanisms will facilitate the determination of how such transfer occurs. Genome editing will allow pinpointing which genes involved in specific transfer mechanisms are the critical determinants of miRNA exchange between cells. The long-term goal is to address questions about how cells communicate with each other. Successful completion of the proposed work will identify miRNAs that mediate cell-cell communication and generate a new set of resources and tools for investigating the relationship between cells in many different models of human biology.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
