Abstract
Majority of disease-modifying therapeutic targets are restricted to the intracellular space and are therefore not druggable using existing biologic modalities. The ability to efficiently deliver macromolecules inside target cells or tissues would greatly expand the current landscape of therapeutic targets for future generations of biologic drugs, but remains challenging. Here we report the use of extracellular vesicles, known as arrestin domain containing protein 1 [ARRDC1]-mediated microvesicles (ARMMs), for packaging and intracellular delivery of a myriad of macromolecules, including the tumor suppressor p53 protein, RNAs, and the genome-editing CRISPR-Cas9/guide RNA complex. We demonstrate selective recruitment of these macromolecules into ARMMs. When delivered intracellularly via ARMMs, these macromolecules are biologically active in recipient cells. P53 delivered via ARMMs induces DNA damage-dependent apoptosis in multiple tissues in mice. Together, our results provide proof-of-principle demonstration that ARMMs represent a highly versatile platform for packaging and intracellular delivery of therapeutic macromolecules.
Highlights
Majority of disease-modifying therapeutic targets are restricted to the intracellular space and are not druggable using existing biologic modalities
In this study we explore the ability of ARRDC1]-mediated microvesicles (ARMMs) to efficiently package and deliver diverse classes of cargo macromolecules, including p53 protein, RNA molecules, and the CRISPR-Cas9 system uses an RNA-guided DNA nuclease (Cas9)/ gRNA complex
During ARMMs biogenesis, ARRDC1 itself is incorporated into the vesicles[9]; we reasoned that fusing p53 to ARRDC1 may enable p53 protein to be incorporated into ARMMs
Summary
Majority of disease-modifying therapeutic targets are restricted to the intracellular space and are not druggable using existing biologic modalities. These extracellular vesicles (EVs) contain functional molecules such as proteins and RNAs, which can be taken up by recipient cells to mediate intercellular communication[2, 3] Because of their ability to carry and transfer bioactive molecules, EVs have been proposed as a new vehicle for therapeutic delivery[4, 5]. This allows controlled production of ARMMs using modern biological manufacturing methods Endogenous proteins such as cell surface receptors are actively recruited into ARMMs and can be delivered into recipient cells to initiate intercellular communication[10], suggesting that the exogenous cargo molecules may be packaged and delivered via ARMMs. In this study we explore the ability of ARMMs to efficiently package and deliver diverse classes of cargo macromolecules, including p53 protein, RNA molecules, and the CRISPR-Cas9/ gRNA complex. Our data provide a proof-of-principle demonstration for the utility of ARMMs to package and deliver a variety of bioactive macromolecules
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
