Cationic polymers for non-viral gene delivery to human T cells

J Control Release. 2018 Jul 28;282:140-147. doi: 10.1016/j.jconrel.2018.02.043. Epub 2018 Mar 5.

Abstract

The clinical success of chimeric antigen receptor (CAR) T cell immunotherapy in treating multiple blood cancers has created a need for efficient methods of ex vivo gene delivery to primary human T cells for cell engineering. Here, we synthesize and evaluate a panel of cationic polymers for gene delivery to both cultured and primary human T cells. We show that a subset of comb- and sunflower-shaped pHEMA-g-pDMAEMA polymers can mediate transfection with efficiencies up to 50% in the Jurkat human T cell line with minimal concomitant toxicity (>90% viability). We then optimize primary human T cell transfection conditions including activation time, cell density, DNA dose, culture media, and cytokine treatment. We demonstrate transfection of both CD4+ and CD8+ primary human T cells with messenger RNA and plasmid DNA at efficiencies up to 25 and 18%, respectively, with similarly high viability.

Keywords: Cationic polymers; Nonviral gene delivery; T lymphocytes.

Publication types

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

MeSH terms

  • Cell Survival / drug effects
  • DNA / administration & dosage*
  • DNA / genetics
  • Drug Carriers / chemistry*
  • Drug Carriers / metabolism
  • Drug Carriers / toxicity
  • Humans
  • Jurkat Cells
  • Methacrylates / chemistry*
  • Methacrylates / metabolism
  • Methacrylates / toxicity
  • Nylons / chemistry*
  • Nylons / metabolism
  • Nylons / toxicity
  • Plasmids / administration & dosage
  • Plasmids / genetics
  • Polyhydroxyethyl Methacrylate / chemistry*
  • Polyhydroxyethyl Methacrylate / metabolism
  • Polyhydroxyethyl Methacrylate / toxicity
  • RNA, Messenger / administration & dosage*
  • RNA, Messenger / genetics
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / metabolism*
  • Transfection / methods*

Substances

  • Drug Carriers
  • Methacrylates
  • Nylons
  • RNA, Messenger
  • poly(2-(dimethylamino)ethyl methacrylate)
  • Polyhydroxyethyl Methacrylate
  • DNA