RT Journal Article
SR Electronic
T1 Engineering of extracellular vesicles for display of protein biotherapeutics
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.14.149823
DO 10.1101/2020.06.14.149823
A1 Dhanu Gupta
A1 Oscar P.B Wiklander
A1 André Görgens
A1 Mariana Conceição
A1 Giulia Corso
A1 Xiuming Liang
A1 Yiqi Seow
A1 Sriram Balsu
A1 Ulrika Felldin
A1 Beklem Bostancioglu
A1 Yi Xin Fiona Lee
A1 Justin Hean
A1 Imre Mäger
A1 Thomas C. Roberts
A1 Manuela Gustafsson
A1 Dara K Mohammad
A1 Helena Sork
A1 Alexandra Bäcklund
A1 C.I. Edvard Smith
A1 Matthew J.A. Wood
A1 Roosmarijn Vandenbroucke
A1 Joel Z. Nordin
A1 Samir EL Andaloussi
YR 2020
UL http://biorxiv.org/content/early/2020/06/15/2020.06.14.149823.abstract
AB Extracellular vesicles (EVs) have recently emerged as a highly promising cell-free bio-therapeutics. While a range of engineering strategies have been developed to functionalize the EV surface, current approaches fail to address the limitations associated with endogenous surface display, pertaining to the heterogeneous display of commonly used EV-loading moieties among different EV subpopulations. Here we present a novel engineering platform to display multiple protein therapeutics simultaneously on the EV surface. As proof-of-concept, we screened multiple endogenous display strategies for decorating the EV surface with cytokine binding domains derived from tumor necrosis factor receptor 1 (TNFR1) and interleukin 6 signal transducer (IL6ST), which can act as decoys for the pro-inflammatory cytokines TNFα and IL6, respectively. Combining synthetic biology and systematic screening of loading moieties, resulted in a three-component system which increased the display and decoy activity of TNFR1 and IL6ST, respectively. Further, this system allowed for combinatorial functionalization of two different receptors on the same EV surface. These cytokine decoy EVs significantly ameliorated disease phenotypes in three different inflammatory mouse models for systemic inflammation, neuroinflammation, and intestinal inflammation. Importantly, significantly improved in vitro and in vivo efficacy of these engineered EVs was observed when compared directly to clinically approved biologics targeting the IL6 and TNFα pathways.Competing Interest StatementMJAW and SEA are founders of, and consultants for, Evox Therapeutics. DG, OW, AG & JZN are consultant for Evox Therapeutics. DG, OW, AG, JZN, MJAW and SEA are shareholders in Evox Therapeutics. The remaining authors declare no conflicts of interest.