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Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers
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How Do Biomolecules Cross the Cell Membrane?

Dehua Pei1

  • 1Department of Chemistry and Biochemistry and Ohio State Biochemistry Program, The Ohio State University, 484 West 12th Avenue, Columbus, Ohio 43210, United States.

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This summary is machine-generated.

A novel vesicle budding-and-collapse (VBC) mechanism explains how cell-penetrating peptides and toxins cross endosomal membranes. This discovery addresses a key challenge in drug delivery and protein transport across cell membranes.

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Area of Science:

  • Cell Biology
  • Membrane Transport
  • Drug Delivery Systems

Background:

  • Biomolecules like proteins and nucleic acids struggle to cross cell membranes passively.
  • Endosomal escape of internalized molecules and protein secretion mechanisms (TAT, UPS) remain poorly understood.
  • This knowledge gap hinders the development of effective drug delivery systems.

Purpose of the Study:

  • To introduce and elucidate the vesicle budding-and-collapse (VBC) mechanism for membrane translocation.
  • To demonstrate VBC's role in endosomal escape of cell-penetrating peptides and toxins.
  • To propose VBC as a unifying mechanism for various protein transport systems.

Main Methods:

  • Utilized giant unilamellar vesicles and live mammalian cells for experiments.
  • Employed pH-sensitive (pHAb) and pH-insensitive (TopFluor) dyes to track biomolecule and lipid trafficking.
  • Applied confocal microscopy for real-time monitoring of intracellular processes.
  • Used a kinase inhibitor to enlarge endosomes and visualize VBC intermediates.

Main Results:

  • Demonstrated that cell-penetrating peptides and bacterial toxins utilize the VBC mechanism for endosomal escape.
  • Observed topological translocation across membranes via VBC, without physical passage through the membrane.
  • Visualized endosomal escape intermediates during the VBC process.
  • Provided evidence for VBC's role in both endosomal escape and protein export systems.

Conclusions:

  • The VBC mechanism offers a new paradigm for understanding membrane translocation of diverse biomolecules.
  • This finding resolves a long-standing mystery in cell biology regarding endosomal escape.
  • VBC provides crucial insights for designing improved drug delivery vehicles and understanding protein transport systems.