Direct Cytosolic Delivery of Proteins through Coengineering of Proteins and Polymeric Delivery Vehicles

J Am Chem Soc. 2020 Mar 4;142(9):4349-4355. doi: 10.1021/jacs.9b12759. Epub 2020 Feb 25.

Abstract

Nanocarrier-mediated protein delivery is a promising strategy for fundamental research and therapeutic applications. However, the efficacy of the current platforms for delivery into cells is limited by endosomal entrapment of delivered protein cargo with concomitantly inefficient access to the cytosol and other organelles, including the nucleus. We report here a robust, versatile polymeric-protein nanocomposite (PPNC) platform capable of efficient (≥90%) delivery of proteins to the cytosol. We synthesized a library of guanidinium-functionalized poly(oxanorborneneimide) (PONI) homopolymers with varying molecular weights to stabilize and deliver engineered proteins featuring terminal oligoglutamate "E-tags". The polymers were screened for cytosolic delivery efficiency using imaging flow cytometry with cytosolic delivery validated using confocal microscopy and activity of the delivered proteins demonstrated through functional assays. These studies indicate that the PPNC platform provides highly effective and tunable cytosolic delivery over a wide range of formulations, making them robust agents for therapeutic protein delivery.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Drug Carriers / chemical synthesis
  • Drug Carriers / metabolism*
  • Guanidines / chemical synthesis
  • Guanidines / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Imides / chemical synthesis
  • Imides / metabolism
  • Integrases / metabolism*
  • Luminescent Proteins / metabolism*
  • Nanocomposites / chemistry
  • Polyglutamic Acid / metabolism*
  • Polymers / chemical synthesis
  • Polymers / metabolism*
  • Protein Engineering
  • Red Fluorescent Protein

Substances

  • Drug Carriers
  • Guanidines
  • Imides
  • Luminescent Proteins
  • Polymers
  • Polyglutamic Acid
  • Cre recombinase
  • Integrases