Low-frequency ultrasound-mediated cytokine transfection enhances T cell recruitment at local and distant tumor sites

Tali Ilovitsh, Yi Feng, Josquin Foiret, Azadeh Kheirolomoom, Hua Zhang, Elizabeth S. Ingham, Asaf Ilovitsh, Spencer K. Tumbale, Brett Z. Fite, Bo Wu, Marina N. Raie, Nisi Zhang, Aris J. Kare, Michael Chavez, Lei S. Qi, Gadi Pelled, Dan Gazit, Ophir Vermesh, Idan Steinberg, Sanjiv S. GambhirKatherine W. Ferrara

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

Robust cytotoxic T cell infiltration has proven to be difficult to achieve in solid tumors. We set out to develop a flexible protocol to efficiently transfect tumor and stromal cells to produce immune-activating cytokines, and thus enhance T cell infiltration while debulking tumor mass. By combining ultrasound with tumor-targeted microbubbles, membrane pores are created and facilitate a controllable and local transfection. Here, we applied a substantially lower transmission frequency (250 kHz) than applied previously. The resulting microbubble oscillation was significantly enhanced, reaching an effective expansion ratio of 35 for a peak negative pressure of 500 kPa in vitro. Combining low-frequency ultrasound with tumor-targeted microbubbles and a DNA plasmid construct, 20% of tumor cells remained viable, and ∼20% of these remaining cells were transfected with a reporter gene both in vitro and in vivo. The majority of cells transfected in vivo were mucin 1+/ CD45 tumor cells. Tumor and stromal cells were then transfected with plasmid DNA encoding IFN-β, producing 150 pg/106 cells in vitro, a 150-fold increase compared to no-ultrasound or no-plasmid controls and a 50-fold increase compared to treatment with targeted microbubbles and ultrasound (without IFN-β). This enhancement in secretion exceeds previously reported fourfold to fivefold increases with other in vitro treatments. Combined with intraperitoneal administration of checkpoint inhibition, a single application of IFN-β plasmid transfection reduced tumor growth in vivo and recruited efficacious immune cells at both the local and distant tumor sites.

Original languageEnglish
Pages (from-to)12674-12685
Number of pages12
JournalProceedings of the National Academy of Sciences of the United States of America
Volume117
Issue number23
DOIs
StatePublished - 9 Jun 2020
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.

Keywords

  • Microbubble
  • Transfection
  • Ultrasound

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