Cell Surface Labeling by Engineered Extracellular Vesicles

Nicklas Hamilton, Natalie M. Claudio, Randall J. Armstrong, Ferdinando Pucci

Research output: Contribution to journalArticlepeer-review

Abstract

Extracellular vesicles (EVs) can mediate local and long-range intercellular communication via cell surface signaling. In order to perform in vivo studies of unmanipulated, endogenously released EVs, sensitive but stringent approaches able to detect EV–cell surface interactions are needed. However, isolation and reinfusion of EVs can introduce biases. A rigorous way to study EVs in vivo is by genetically engineering membrane-bound reporters into parental cells. Still, the amount of reporter molecules that EVs can carry is relatively small, and thus, the sensitivity of the approach is suboptimal. This work addresses this issue by engineering EVs to display a membrane-bound form of Sortase A (SrtA), a bacterial transpeptidase that can catalyze the transfer of reporter molecules on the much bigger surface of EV-binding cells. SrtA design and reaction requirements are optimized and validated. Efficient in vitro labeling of EV-binding cells is achieved, even in the presence of only one N-terminal glycine on cell surface proteins. As compared to indirect labeling of EV-binding cells (e.g., using CD63-GFP fusion), the SrtA-based approach shows 1–2 log increase in sensitivity, depending on the EV source. This novel approach will be useful to identify and study the full set of host cells interacting with native EVs in vivo.

Original languageEnglish (US)
Article number2000007
JournalAdvanced Biosystems
Volume4
Issue number12
DOIs
StatePublished - Dec 2020

Keywords

  • cell surface labeling
  • exosomes
  • extracellular vesicles
  • intercellular communication
  • squamous cell carcinoma

ASJC Scopus subject areas

  • Biomaterials
  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology(all)

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