Enhanced‐mRNA Delivery Using Ultrasound‐Delivered Anchors for Bioorthogonal Ligation
Abstract
Abstract Therapeutic nucleic acid delivery has many potential applications, but it remains challenging to target extrahepatic tissues in a flexible and image‐guided manner. To address this issue, we report a bioorthogonal pre‐targeting strategy that uses focused ultrasound (FUS) to promote the delivery of mRNA‐loaded lipid nanoparticles (mRNA‐LNP). We synthesized amphiphilic click reactive anchors (ACRAs) consisting of a phospholipid PEG‐conjugate functionalized with trans‐cyclooctene (TCO) or its companion reactive partner methyltetrazine (mTz), producing ACRA‐TCO and ACRA‐mTz. ACRA derivatives were screened for cellular activity, yielding functionalized DOPE‐PEG (1,2‐dioleoyl‐sn‐glycero‐3‐phosphoethanolamine‐N‐(polyethylene glycol)) derivatives outperforming those containing saturated lipid or branched PEG. Nanobubbles encapsulating ultrasound‐responsive gas delivered ACRA‐TCO to targeted cells and tissues using FUS. This pre‐targeting promoted the subsequent delivery of mRNA‐LNP functionalized with companion ACRA‐mTz. Ultrasound pre‐targeting enhanced the accumulation of mTz‐functionalized nanoparticles in cell cultures and mice by 75% and up to 3.6‐fold, respectively, and increased gene expression using mRNA‐LNP in vivo. Microbubbles loaded with ACRA roughly doubled mRNA delivery to the heart, while ultrasound alone did not. Taken together, this report presents a modular, ultrasound‐enabled strategy for enhancing nucleic acid delivery in targeted tissues.
Article Details
Authors (20)
Emilio Di Ianni
Department of Neurology Molecular Neurogenetics Unit Massachusetts General Hospital Harvard Medical School Charlestown Massachusetts USA
Jueun Jeon
Center for Systems Biology
Sedra Mohammadi
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Huiyu Hu
Jeremy M. Quintana
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Chanseo Lee
Edwina Abou Haidar
Department of Neurology Molecular Neurogenetics Unit Massachusetts General Hospital Harvard Medical School Charlestown Massachusetts USA
Marie Goemans
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Ayrton Zargani‐Piccardi
Department of Neurology Molecular Neurogenetics Unit Massachusetts General Hospital Harvard Medical School Charlestown Massachusetts USA
Mohammed Mahamdeh
Department of Medicine Cardiovascular Research Center Cardiology Division Massachusetts General Hospital Harvard Medical School Boston Massachusetts USA
Iván Coto Hernández
Institute for Innovation in Imaging Massachusetts General Hospital Harvard Medical School Boston Massachusetts USA
Victoria E. Rodriguez
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Yirong Zhou
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Aaron Aguirre
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Shiaulou Yuan
Department of Medicine Cardiovascular Research Center Cardiology Division Massachusetts General Hospital Harvard Medical School Boston Massachusetts USA
Thomas S.C. Ng
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA
Koen Breyne
Department of Neurology Molecular Neurogenetics Unit Massachusetts General Hospital Harvard Medical School Charlestown Massachusetts USA
Hakho Lee
Center for Systems Biology
Xandra O. Breakefield
Department of Neurology Molecular Neurogenetics Unit Massachusetts General Hospital Harvard Medical School Charlestown Massachusetts USA
Miles A. Miller
Center for Systems Biology Massachusetts General Hospital Boston Massachusetts USA