Extracellular RNA (exRNA) describes RNA species present outside of the cells in which they were transcribed. Carried within extracellular vesicles, lipoproteins, and protein complexes, exRNAs are protected from ubiquitous RNA-degrading enzymes. exRNAs may be found in the environment or, in multicellular organisms, within the tissues or biological fluids such as venous blood, saliva, breast milk, urine, semen, menstrual blood, and vaginal fluid. Although their biological function is not fully understood, exRNAs have been proposed to play a role in a variety of biological processes including syntrophy, intercellular communication, and cell regulation. The United States National Institutes of Health (NIH) published in 2012 a set of Requests for Applications (RFAs) for investigating extracellular RNA biology. Funded by the NIH Common Fund, the resulting program was collectively known as the Extracellular RNA Communication Consortium (ERCC). The ERCC was renewed for a second phase in 2019.
Background
Both prokaryotic and eukaryotic cells are known to release RNA, and this release can be passive or active. The Endosomal Sorting Complex Required for Transport (ESCRT) machinery was previously considered as a possible mechanism for RNA secretion from the cell, but more recently research studying microRNA secretion in human embryonic kidney cells and Cercopithecus aethiops kidney cells identified neutral sphingomyelinase 2 (nSMase2), an enzyme involved in ceramide biosynthesis, as a regulator of microRNA secretion levels. ExRNAs are often found packaged within vesicles such as exosomes, ectosomes, prostasomes, microvesicles, and apoptotic bodies. Although RNAs can be excreted from the cell without an enveloping container, ribonucleases present in extracellular environments would eventually degrade the molecule.
Types
Extracellular RNA should not be viewed as a category describing a set of RNAs with a specific biological function or belonging to a particular RNA family. Similar to the term "non-coding RNA", "extracellular RNA" defines a group of several types of RNAs whose functions are diverse, yet they share a common attribute which, in the case of exRNAs, is existence in an extracellular environment. The following types of RNA have been found outside the cell: Messenger RNA (mRNA) Transfer RNA (tRNA) MicroRNA (miRNA) Small interfering RNA (siRNA) Long non-coding RNA (lncRNA)
Though prevalent inside of the cell, ribosomal RNA (rRNA) does not seem to be a common exRNA. Efforts by Valadi et al. to characterize exosomal RNA using the Agilent Bioanalyzer technology showed little to no trace of 18S and 28S rRNA in exosomes secreted by MC/9 murine mast cells, and similar conclusions were made by Skog et al. for rRNA in gliobastoma microvesicles.
Function
To function or even survive as full-length RNA in extracellular environments, exRNA must be protected from digestion by RNases. This requirement does not apply to prokaryotic syntrophy, where digested nucleotides are recycled. exRNA can be shielded from RNases by RNA binding proteins (RBPs), on their own or within/associated with lipoprotein particles and extracellular vesicles. Extracellular vesicles in particular are thought to be a way to transport RNA between cells, in a process that may be general or highly specific, for example, due to incorporation of markers of the parent cell that may be recognized by receptors on the recipient cell. Biochemical evidence supports the idea that exRNA uptake is a common process, suggesting new pathways for intercellular communication. As a result, the presence, absence, and relative abundance of certain exRNAs can be correlated with changes in cellular signaling and may indicate specific disease states.