2. Academic Validation
  3. Orthogonal Design of Experiments for Optimization of Lipid Nanoparticles for mRNA Engineering of CAR T Cells

Orthogonal Design of Experiments for Optimization of Lipid Nanoparticles for mRNA Engineering of CAR T Cells

  • Nano Lett. 2022 Jan 12;22(1):533-542. doi: 10.1021/acs.nanolett.1c02503.
Margaret M Billingsley 1,  Alex G Hamilton 1,  David Mai 1 2,  Savan K Patel 1,  Kelsey L Swingle 1,  Neil C Sheppard 2 3,  Carl H June 2 3,  Michael J Mitchell 1 2 4 5 6
Affiliations

Affiliations

  • 1 Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 2 Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 3 Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 4 Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 5 Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 6 Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
PMID: 34669421 DOI: 10.1021/acs.nanolett.1c02503
Abstract

Viral engineered chimeric antigen receptor (CAR) T cell therapies are potent, targeted Cancer immunotherapies, but their permanent CAR expression can lead to severe adverse effects. Nonviral messenger RNA (mRNA) CAR T cells are being explored to overcome these drawbacks, but electroporation, the most common T Cell Transfection method, is limited by cytotoxicity. As a potentially safer nonviral delivery strategy, here, sequential libraries of ionizable lipid nanoparticle (LNP) formulations with varied excipient compositions were screened in comparison to a standard formulation for improved mRNA delivery to T cells with low cytotoxicity, revealing B10 as the top formulation with a 3-fold increase in mRNA delivery. When compared to electroporation in primary human T cells, B10 LNPs induced comparable CAR expression with reduced cytotoxicity while demonstrating potent Cancer cell killing. These results demonstrate the impact of excipient optimization on LNP performance and support B10 LNPs as a potent mRNA delivery platform for T cell engineering.

Keywords

CAR T; T cell engineering; lipid nanoparticles; mRNA delivery.

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Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-148842
    99.01%, ionizable lipid