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Cellular Uptake of Phase‐Separating Peptide Coacervates

Authors :
Anastasia Shebanova
Quentin Moana Perrin
Kexin Zhu
Sushanth Gudlur
Zilin Chen
Yue Sun
Congxi Huang
Zhi Wei Lim
Evan Angelo Mondarte
Ruoxuan Sun
Sierin Lim
Jing Yu
Yansong Miao
Atul N. Parikh
Alexander Ludwig
Ali Miserez
Source :
Advanced Science, Vol 11, Iss 42, Pp n/a-n/a (2024)
Publication Year :
2024
Publisher :
Wiley, 2024.

Abstract

Abstract Peptide coacervates self‐assembling via liquid‐liquid phase separation are appealing intracellular delivery vehicles of macromolecular therapeutics (proteins, DNA, mRNA) owing to their non‐cytotoxicity, high encapsulation capacity, and efficient cellular uptake. However, the mechanisms by which these viscoelastic droplets cross the cellular membranes remain unknown. Here, using multimodal imaging, data analytics, and biochemical inhibition assays, we identify the key steps by which droplets enter the cell. We find that the uptake follows a non‐canonical pathway and instead integrates essential features of macropinocytosis and phagocytosis, namely active remodeling of the actin cytoskeleton and appearance of filopodia‐like protrusions. Experiments using giant unilamellar vesicles show that the coacervates attach to the bounding membrane in a charge‐ and cholesterol‐dependent manner but do not breach the lipid bilayer barrier. Cell uptake in the presence of small molecule inhibitors – interfering with actin and tubulin polymerization – confirm the active role of cytoskeleton remodeling, most prominently evident in electron microscopy imaging. These findings suggest a peculiar internalization mechanism for viscoelastic, glassy coacervate droplets combining features of non‐specific uptake of fluids by macropinocytosis and particulate uptake of phagocytosis. The broad implications of this study will enable to enhance the efficacy and utility of coacervate‐based strategies for intracellular delivery of macromolecular therapeutics.

Details

Language :
English
ISSN :
21983844
Volume :
11
Issue :
42
Database :
Directory of Open Access Journals
Journal :
Advanced Science
Publication Type :
Academic Journal
Accession number :
edsdoj.164071ca5274ec38706c5da76084983
Document Type :
article
Full Text :
https://doi.org/10.1002/advs.202402652