Type
Text
Type
Dissertation
Advisor
Gingeras, Thomas R. | Jackson, David | VanAelst, Linda | Thomsen, Gerald | Taylor, Douglas.
Date
2015-05-01
Keywords
Exosomes, Extracellular vesicles, Genomics, Inter-cellular communication, Non-coding RNA, RNA-Seq | Genetics
Department
Department of Genetics.
Language
en_US
Source
This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.
Identifier
http://hdl.handle.net/11401/77613
Publisher
The Graduate School, Stony Brook University: Stony Brook, NY.
Format
application/pdf
Abstract
Proper functioning of a multi-cellular organism requires well-coordinated intra- and inter-cellular communication among its cells. Mammalian cells have well established modes of inter-cellular communication, which include cell-cell contact via gap junctions, synaptic transmission, and paracrine and endocrine communication through secreted molecules. Extracellular vesicles (EV) serve as a vehicle for transfer of biomolecules between cells. This study aims to establish if EV mediated transfer of encapsulated RNA represents a novel mode of inter-cellular communication. At the outset, we developed and validated a novel and efficient approach of isolation of EVs. Comprehensive characterization of EV RNA derived from several cell types revealed the diversity of small RNA cargo, and suggested non-random and cell type specific sorting of RNA molecules into EVs. Further analysis revealed gene family specific fragmentation patterns of non-coding RNAs detected within EVs. Next, we explored the temporal dynamics, spatial localization, and integrity of EV RNA upon transfer into another cell. We further demonstrated cell state dependent dynamicity of EV RNA cargo, as well as cell type specific molecular responses, when different cells were exposed to the same EV stimuli, underscoring the context dependent interpretation of the complex EV messages. Finally, as a specific example of EV RNA functionality, we reported the possible involvement of a 31 nucleotide processed fragment of RNY5, one of the most abundant and enriched RNA components of cancer cell derived EVs, in selectively inducing cell death in primary cells of diverse developmental origins and identified an eight nucleotide motif crucial for its functionality. The transfer of processed, functional RNY5 fragments through EVs hints at the under-appreciated role of EV RNA in cancer cell microenvironment. Thus, our study supports the hypothesis that EV mediated transfer of RNA represents a novel mode of inter-cellular communication, through which cells may shape the transcriptional landscape of another cell in their microenvironment. | 182 pages
Recommended Citation
Chakrabortty, Sudipto Kumar, "Intercellular transfer of functional RNA through extracellular vesicles is a medium of communication between cells" (2015). Stony Brook Theses and Dissertations Collection, 2006-2020 (closed to submissions). 3408.
https://commons.library.stonybrook.edu/stony-brook-theses-and-dissertations-collection/3408