Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

Samagya Banskota; Aditya Raguram; Susie Suh; Samuel W Du; Jessie R Davis; Elliot H Choi; Xiao Wang; Sarah C Nielsen; Gregory A Newby; Peyton B Randolph; Mark J Osborn; Kiran Musunuru; Krzysztof Palczewski; David R Liu

doi:10.1016/j.cell.2021.12.021

Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

Cell. 2022 Jan 20;185(2):250-265.e16. doi: 10.1016/j.cell.2021.12.021. Epub 2022 Jan 11.

Authors

Affiliations

¹ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.
² Gavin Herbert Eye Institute, Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, CA, USA; Department of Pharmacology, Case Western Reserve University, Cleveland, OH, USA.
³ Gavin Herbert Eye Institute, Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, CA, USA; Department of Physiology and Biophysics, University of California, Irvine, CA, USA.
⁴ Cardiovascular Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA; Division of Cardiovascular Medicine, Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA; Department of Genetics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
⁵ Department of Pediatrics, Division of Blood and Marrow Transplant and Cellular Therapy, University of Minnesota, Minneapolis, MN, USA.
⁶ Gavin Herbert Eye Institute, Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, CA, USA; Department of Physiology and Biophysics, University of California, Irvine, CA, USA; Department of Chemistry, University of California, Irvine, CA, USA; Department of Molecular Biology and Biochemistry, University of California, Irvine, CA, USA.
⁷ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA. Electronic address: [email protected].

Abstract

Methods to deliver gene editing agents in vivo as ribonucleoproteins could offer safety advantages over nucleic acid delivery approaches. We report the development and application of engineered DNA-free virus-like particles (eVLPs) that efficiently package and deliver base editor or Cas9 ribonucleoproteins. By engineering VLPs to overcome cargo packaging, release, and localization bottlenecks, we developed fourth-generation eVLPs that mediate efficient base editing in several primary mouse and human cell types. Using different glycoproteins in eVLPs alters their cellular tropism. Single injections of eVLPs into mice support therapeutic levels of base editing in multiple tissues, reducing serum Pcsk9 levels 78% following 63% liver editing, and partially restoring visual function in a mouse model of genetic blindness. In vitro and in vivo off-target editing from eVLPs was virtually undetected, an improvement over AAV or plasmid delivery. These results establish eVLPs as promising vehicles for therapeutic macromolecule delivery that combine key advantages of both viral and nonviral delivery.

Keywords: base editing; genome editing; in vivo delivery; ribonucleoproteins; therapeutic gene editing; virus-like particles.

Publication types

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

MeSH terms

Animals
Base Sequence
Blindness / genetics
Blindness / therapy
Brain / metabolism
DNA / metabolism
Disease Models, Animal
Drug Delivery Systems*
Fibroblasts / metabolism
Gene Editing
Genetic Engineering*
HEK293 Cells
Humans
Liver / pathology
Mice
Mice, Inbred C57BL
Proprotein Convertase 9 / metabolism
Proteins / therapeutic use*
Retinal Pigment Epithelium / pathology
Retroviridae
Virion / genetics*
Virion / ultrastructure
Vision, Ocular

Substances

Proteins
DNA
Proprotein Convertase 9

Abstract

Publication types

MeSH terms

Substances

Grants and funding