2. Academic Validation
  3. Branched endosomal disruptor (BEND) lipids mediate delivery of mRNA and CRISPR-Cas9 ribonucleoprotein complex for hepatic gene editing and T cell engineering

Branched endosomal disruptor (BEND) lipids mediate delivery of mRNA and CRISPR-Cas9 ribonucleoprotein complex for hepatic gene editing and T cell engineering

  • Nat Commun. 2025 Jan 24;16(1):996. doi: 10.1038/s41467-024-55137-6.
Marshall S Padilla 1,  Kaitlin Mrksich 1,  Yiming Wang 1 2 3,  Rebecca M Haley 1,  Jacqueline J Li 1,  Emily L Han 1,  Rakan El-Mayta 1 4,  Emily H Kim 1,  Sofia Dias 1 4 5 6,  Ningqiang Gong 1,  Sridatta V Teerdhala 1,  Xuexiang Han 1,  Vivek Chowdhary 7,  Lulu Xue 1,  Zain Siddiqui 1,  Hannah M Yamagata 1,  Dongyoon Kim 1,  Il-Chul Yoon 1,  James M Wilson 7,  Ravi Radhakrishnan 1 2 3,  Michael J Mitchell 8 9 10 11 12 13 14
Affiliations

Affiliations

  • 1 Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, 19104, USA.
  • 2 Penn Institute for Computational Science, University of Pennsylvania, Philadelphia, PA, 19104, USA.
  • 3 Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA, 19104, USA.
  • 4 Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, 4200-135, Portugal.
  • 5 Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, 4050-313, Portugal.
  • 6 Instituto de Engenharia Biomédica, Universidade do Porto, Porto, 4200-135, Portugal.
  • 7 Gene Therapy Program, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA.
  • 8 Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
  • 9 Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
  • 10 Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
  • 11 Penn Institute for RNA Innovation, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
  • 12 Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
  • 13 Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
  • 14 Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. mjmitch@seas.upenn.edu.
PMID: 39856035 DOI: 10.1038/s41467-024-55137-6
Abstract

Lipid nanoparticles (LNPs) are the preeminent non-viral drug delivery vehicle for mRNA-based therapies. Immense effort has been placed on optimizing the ionizable lipid (IL) structure, which contains an amine core conjugated to lipid tails, as small molecular adjustments can result in substantial changes in the overall efficacy of the resulting LNPs. However, despite some advancements, a major barrier for LNP delivery is endosomal escape. Here, we develop a platform for synthesizing a class of branched ILs that improve endosomal escape. These compounds incorporate terminally branched groups that increase hepatic mRNA and ribonucleoprotein complex delivery and gene editing efficiency as well as T Cell Transfection compared to non-branched lipids. Through an array of complementary experiments, we determine that our lipid architecture induces greater endosomal penetration and disruption. This work provides a scheme to generate a class of ILs for both mRNA and protein delivery.

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