In vivo selection of synthetic nucleocapsids for tissue targeting

Abstract

Controlling the biodistribution of protein- and nanoparticle-based therapeutic formulations remains challenging. In vivo library selection is an effective method for identifying constructs that exhibit desired distribution behavior; library variants can be selected based on their ability to localize to the tissue or compartment of interest despite complex physiological challenges. Here, we describe further development of an in vivo library selection platform based on self-assembling protein nanoparticles encapsulating their own mRNA genomes (synthetic nucleocapsids or synNCs). We tested two distinct libraries: a low-diversity library composed of synNC surface mutations (45 variants) and a high-diversity library composed of synNCs displaying miniproteins with binder-like properties (6.2 million variants). While we did not identify any variants from the low-diversity surface library that yielded therapeutically relevant changes in biodistribution, the high-diversity miniprotein display library yielded variants that shifted accumulation toward lungs or muscles in just two rounds of in vivo selection. Our approach should contribute to achieving specific tissue homing patterns and identifying targeting ligands for diseases of interest.

Document Details

Document Type
Pub Defense Publication
Publication Date
Nov 08, 2023
Source ID
10.1073/pnas.2306129120

Entities

People

  • Audrey Olshefsky
  • Catherine Treichel
  • Christian Richardson
  • David Baker
  • Derrick R. Hicks
  • Elizabeth M. Leaf
  • Gabriel J. Rocklin
  • Gabriel L. Butterfield
  • Gargi Kher
  • Halli Benasutti
  • Jeffrey S Chamberlain
  • Justin Decarreau
  • Kefan Song
  • Lauren Carter
  • Marc J. Lajoie
  • Meilyn Sylvestre
  • Neil P. King
  • Sharon Ke
  • Suzie H. Pun

Organizations

  • Defense Threat Reduction Agency
  • Gates Foundation
  • National Cancer Institute
  • National Institute of Allergy and Infectious Diseases
  • University of Washington

Tags

Fields of Study

  • Biology

Readers

  • Molecular Genetics
  • Nanocomposite Materials Science

Technology Areas

  • Biotechnology
  • Biotechnology - Cancer Biotech