Drug Delivery via Cell Membrane Fusion Using Lipopeptide Modified Liposomes

Abstract

Efficient delivery of drugs to living cells is still a major challenge. Currently, most methods rely on the endocytotic pathway resulting in low delivery efficiency due to limited endosomal escape and/or degradation in lysosomes. Here, we report a new method for direct drug delivery into the cytosol of live cells in vitro and invivo utilizing targeted membrane fusion between liposomes and live cells. A pair of complementary coiled-coil lipopeptides was embedded in the lipid bilayer of liposomes and cell membranes respectively, resulting in targeted membrane fusion with concomitant release of liposome encapsulated cargo including fluorescent dyes and the cytotoxic drug doxorubicin. Using a wide spectrum of endocytosis inhibitors and endosome trackers, we demonstrate that the major site of cargo release is at the plasma membrane. This method thus allows for the quick and efficient delivery of drugs and is expected to have many invitro, ex vivo, and invivo applications.

Membrane Fusion Mediated Intracellular Delivery of Lipid Bilayer Coated Mesoporous Silica Nanoparticles

“…[27,47] So far, this approach was used to deliver low molecular weight dyes and drugs into the cytosol and nucleus of live cells. To study the scope of this synthetic fusion system, we now were interested to study whether coiled-coil mediated fusion could be used to enhance the intracellular delivery of inorganic nanoparticles like MSNs.…”

“…[27,[50][51][52][53][54] In this study, wortmannin (Wor) was used as a micropinocytosis inhibitor as it blocks PI3-kinase, [50,52,55,56] while genistein (Gen) inhibits tyrosine-phosphorylation of Cav 1 and caveolin-dependent endocytosis. [57][58][59] Furthermore, chlorpromazine (Chl) was used as it blocks clathrin-dependent endocytosis, [51,60,61] and microtubule formation was inhibited by nocodazole (Noc).…”

Glycosylated Cell‐Penetrating Peptides (GCPPs)

“…[3][4][5] More than 20 years ago, it was accidentlyd iscovered that certain protein domains, enriched in cationic amino acids, were responsible for promoting the translocation of some proteins throught he plasma membrane of cells. In 1997, Lebleu et al reportedt hat the peptidef unction of penetrating properties was attributed to the cationic domain, which was called the TATp eptide( TAT [49][50][51][52][53][54][55][56][57] ,R KKRRQRRR). In 1997, Lebleu et al reportedt hat the peptidef unction of penetrating properties was attributed to the cationic domain, which was called the TATp eptide( TAT [49][50][51][52][53][54][55][56][57] ,R KKRRQRRR).…”

“…The group of Wender at Stanford University systematically studied the TAT [49][50][51][52][53][54][55][56][57] peptide sequence and artificial analogues, and demonstrated that between eight and ten cationic amino The cell membrane regulates the exchange of molecules and information with the externale nvironment. However,t his control barrier hinders the delivery of exogenous bioactive molecules that can be appliedt oc orrect cellular malfunctions.…”

Artificial Membrane Fusion Triggered by Strain-Promoted Alkyne–Azide Cycloaddition

“…Only recently have initial examples of in vivo targeted membrane fusion been reported using coiled coil peptides attached onto membrane surfaces. 47,48 Bioorthogonal chemistry presents a promising strategy for attaining this goal due to the selectivity of these reactions in complex biological environments. 49,50 We and others have shown that the derivatization of intact liposomes can be conveniently achieved using bioorthogonal reactions, such as the copper-catalyzed azide-alkyne cycloaddition (CuAAC), 51–56 the strain promoted alkyne-azide cycloaddition (SPAAC), 57–59 and the Staudinger ligation.…”