Cell-surface retained cell-penetrating peptide additives — ASN Events

Cell-surface retained cell-penetrating peptide additives (#35)

Jan Vincent V. Arafiles 1 , Christian P. R. Hackenberger 1
  1. Leibniz-Forschungsinstitut fur Molekulare Pharmakologie, Buch, BERLIN, Germany

We present a simple yet highly effective strategy to achieve a high degree of functional protein delivery into mammalian cells. Inefficient cellular uptake of protein-based drugs remains a major challenge for the biopharmaceutic industry. For example, antibody-drug therapy is restricted to targeting and blocking membrane-bound proteins, leaving a plethora of cancer-related intracellular targets untouched. Hence, we aim to develop efficient, effective, and biocompatible peptide-based protein delivery strategies.

One can achieve protein delivery through chemical conjugation with cell-penetrating peptides (CPPs). These protein-CPP conjugates are reported to enter cells directly via transduction through the cell membrane, albeit requiring high micromolar concentrations (>20 µM).1,2 To address this, we developed the “CPP-additive technology” to facilitate greater direct transductions of protein-CPPs at low micromolar concentrations.3 CPP-additives work by accumulating on the cell membrane, resulting in the reduction of membrane tension to prime entry points of protein-CPP cargoes.

In this presentation, we introduce the second generation of CPP-additives containing a membrane-interacting hydrophobic motif. The hydrophobic CPP-additive enhanced the uptake via direct transduction of a model fluorescent protein compared to the original design. We show that the hydrophobic CPP-additive remains engaged on the cell membrane for a longer time (> 10 min) while creating loose membrane lipid packing defects at an earlier time point (~ 15 s). Finally, we demonstrate various applications for the delivery of biofunctional peptide- and protein cargoes facilitated by the hydrophobic CPP-additives.

  1. Tünnemann, G. et al. FASEB J. 2006. 20, 1775-1784.
  2. Herce, H. D. et al. Nat. Chem. 2017. 9, 762-771.
  3. Schneider, A. et al. Nat. Chem. 2021. 136, 530-539.
  4. Arafiles, J. V. V. et al. In review
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