Project E2 - Identification, analytics, and functional characterization of extracellular vesicles
- Project description
Background: Extracellular vesicles are released by a multitude of different cell types. In particular the endocytosis-generated vesicles (with 40-200 nm in diameter) and the microvesicles, 100-1,000 nm in diameter and directly released from the plasma membrane, are in the focus of current research, based on their enormous potential for diagnostics (liquid biopsy) and for new therapeutic options. Exosomes and other kinds of extracellular vesicles have been established as a new principle of intracellular communication. Various biomolecules such as proteins and RNAs (e.g., microRNAs) are released via extracellular vesicles into all kinds of body fluids, transmitting information and signals and thereby allowing the modulation of gene expression in target cells and tissues. Moreover, this kind of information exchange by extracellular vesicles plays an important role in intracellular communication between parasites and host cells, which contributes to the establishment and pathophysiology of infections (see figure).
Our group focuses on the characterization of extracellular vesicles from various cell lines, parasites (such as Trypanosoma brucei), and bodily fluids such as blood, serum, and plasma. Our expertise and experimental approaches include the stringent purification and characterization of extracellular vesicles from the biosystems mentioned (using differential and density-gradient ultracentrifugation), trancriptome and genome-wide analysis methods including bioinformatic evaluation, and biochemical validation such as via RNA-seq or CLIP-seq in order to map in vivo binding sites of RNA-associated proteins, general RNA biochemistry and biology methods such as RT-qPCR, northern blotting, affinity purification of RNA–protein complexes, cell culture and immunolocalization. Regarding RNA analysis, our special expertise concentrates on circular RNAs (circRNAs), a new class of noncoding, stable RNAs, and their potential as novel biomarkers.
Within this project, we will establish a method platform for the systematic purification and analytics of extracellular vesicles, including exosomes. Here we will apply our expertise to various biological systems and lines of research within our LOEWE center. Based on our method platform, we expect to gain new insight into the intercellular communication between parasites and host cells, thereby exploring the complex networks during infection. This should open up new avenues for the development of highly specific diagnostic biomarkers and therapeutic options.