Abstract 48: A novel method for live cell sorting based on RNA expression levels of cancer biomarkers maintains normal cellular function as confirmed by downstream functional testing

Abstract

Abstract Enrichment of cells within a mixed population has been confined to fixed cells or in the case of live cell sorting to a limited set of cell surface markers which are typically not specific to the target population. Furthermore, cell surface antibodies may activate cell surface receptors altering the state of the cells such that they are no longer representative of the original population. Intracellular staining requires fixation and permeabilization and thus renders the cells unusable in downstream functional assays. Through the use of a unique nanoparticle sequence based detection technology we have shown the ability to detect specific RNA within live cells which does not affect or alter the cells and allows further testing of the target population following the enrichment. Both microRNA and mRNA species have been used to enrich for specific target populations through Fluorescence Activated Cell Sorting (FACS). Sorting cells based on the expression level of any specific RNA allows researchers the ability to utilize targets of biological interest within isolated populations of cells. Since this method is non-destructive to live cells the enriched cells can then be used for downstream functional assays. Here we present data showing the live cell enrichment of a target cell population within a heterogeneous mixture of cells based on their RNA expression levels. RNA probes to miR-221 and miR-222 were generated for the enrichment of invasive cells from the non-invasive. A mixture of breast cancer cells (MD-MB-231 and T47D) were sorted based on their expression levels of the RNA probes through FACS. The high and low expressing sort products were then each subjected to a collagen invasion assay. As expected the fractions which showed high expression of miR-221 and miR-222 showed a high level of invasion through the matrix while the low expressing fractions showed extremely low invasion. Live cell sorting based on RNA levels of cancer biomarkers in a non-destructive manner enables researchers the ability to study the expression of a diverse group of targets beyond the traditional surface markers with the added benefit of utilizing the enriched cells for downstream functional assays. Citation Format: Don Weldon, Victor Koong, Yuko Williams. A novel method for live cell sorting based on RNA expression levels of cancer biomarkers maintains normal cellular function as confirmed by downstream functional testing. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 48. doi:10.1158/1538-7445.AM2014-48