Transduction Methods for Cytosolic Delivery of Proteins and Bioconjugates into Living Cells

Abstract

The human organism and its constituting cells rely on interplay between multiple proteins exerting specific functions. Progress in molecular biotechnologies has facilitated the production of recombinant proteins. When administrated to patients, recombinant proteins can provide important healthcare benefits. To date, most therapeutic proteins must act from the extracellular environment, with their targets being secreted modulators or extracellular receptors. This is because proteins cannot passively diffuse across the plasma membrane into the cytosol. To expand the scope of action of proteins for cytosolic targets (representing more than 40% of the genome) effective methods assisting protein cytosolic entry are being developed. To date, direct protein delivery is extremely tedious and inefficient in cultured cells, even more so in animal models of pathology. Novel techniques are changing this limitation, as recently developed in vitro methods can robustly convey large amount of proteins into cell cultures. Moreover, advances in protein formulation or protein conjugates are slowly, but surely demonstrating efficiency for targeted cytosolic entry of functional protein in vivo in tumor xenograft models. In this review, various methods and recently developed techniques for protein transport into cells are summarized. They are put into perspective to address the challenges encountered during delivery.

Short oligoalanine helical peptides for supramolecular nanopore assembly and protein cytosolic delivery

“…These excellent results encouraged us to test the potential of the helical responsive MP1 for the transport of antibodies, [5][6][7]40,60,61 which was evaluated with mouse monoclonal Mab414 against the nuclear pore complex proteins ( Fig. 6 and Fig.…”

“…[1][2][3][4] However, strongly cationic molecules and surfactants suffer from non-specific binding to membranes and detergent behavior. 1,5,6 Over the last years, different strategies employing cationic synthetic materials 3,7,8 have been developed to transport bioactive macromolecules, 9 which include peptides, [10][11][12][13][14][15][16][17][18][19][20][21] chimeric proteins, 22 polymers, [23][24][25] dynamic covalent bonds, [23][24][25][26] supramolecular caging, 27 transient charge masking, 28 lipid particles, 29,30 polymersomes, 31,32 and highly charged nanoparticles. [33][34][35] Importantly, the presence of anionic lipids, especially in the endosomal compartments, has been proved critical for the correct function of several natural 36,37 and artificial cationic macromolecular carriers.…”

Protein Delivery into the Cell Cytosol using Non-Viral Nanocarriers

“…Alternative transient delivery methods include use of cationic transfection agents such as Lipofectamine 26, 27 or polyethyleneimine (PEI) 28, or through mechanical methods such as electroporation 29 or membrane deformation 30. However, in general approaches involving the delivery of DNA and mRNA do not offer control over the level of protein expressed by the cell or the timeframe over which protein expression occurs, typically resulting in a distribution of expression levels 25, 31. This heterogeneity presents challenges for both fundamental biological studies and therapeutic dosing.…”

Co-delivery of doxorubicin and paclitaxel by reduction/pH dual responsive nanocarriers for osteosarcoma therapy

“…The combination of two or more anticancer drugs with different antitumor mechanisms can enhance the therapeutic effects by the synergy between the different drugs, reduce the adverse effects, and prevent the occurrence of drug resistance. Therefore, it is necessary to develop novel combination therapies to improve clinical osteosarcoma treatment (Fu et al., 2014 ; Jiang et al., 2014 ; He et al., 2016 ; Chiper et al., 2018 ).…”