An extracellular vesicle delivery platform based on the PTTG1IP protein

Carla Martin Perez; Xiuming Liang; Dhanu Gupta; Emily R. Haughton; Mariana Conceição; Imre Mäger; Samir EL Andaloussi; Matthew J.A. Wood; Thomas C. Roberts

Extracellular Vesicle (Dec 2024)

An extracellular vesicle delivery platform based on the PTTG1IP protein

Carla Martin Perez,
Xiuming Liang,
Dhanu Gupta,
Emily R. Haughton,
Mariana Conceição,
Imre Mäger,
Samir EL Andaloussi,
Matthew J.A. Wood,
Thomas C. Roberts

Affiliations

Carla Martin Perez: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK
Xiuming Liang: Biomolecular Medicine, Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden
Dhanu Gupta: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK; Biomolecular Medicine, Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford, OX3 7TY, UK
Emily R. Haughton: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford, OX3 7TY, UK
Mariana Conceição: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford, OX3 7TY, UK; MDUK Oxford Neuromuscular Centre, Oxford, OX3 7TY, UK
Imre Mäger: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK
Samir EL Andaloussi: Biomolecular Medicine, Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden
Matthew J.A. Wood: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford, OX3 7TY, UK; MDUK Oxford Neuromuscular Centre, Oxford, OX3 7TY, UK
Thomas C. Roberts: Department of Paediatrics, University of Oxford, Oxford, OX3 7TY, UK; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford, OX3 7TY, UK; MDUK Oxford Neuromuscular Centre, Oxford, OX3 7TY, UK; Corresponding author. Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus Roosevelt Drive, Headington, Oxford, OX3 7TY, UK.

Journal volume & issue: Vol. 4
p. 100054

Abstract

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Extracellular vesicles (EVs) are promising therapeutic delivery vehicles, although their potential is limited by a lack of efficient engineering strategies to enhance loading and functional cargo delivery. Using an in-house bioinformatics analysis, we identified N-glycosylation as a putative EV-sorting feature. PTTG1IP (a small, N-glycosylated, single-spanning transmembrane protein) was found to be a suitable scaffold for EV loading of therapeutic cargoes, with loading dependent on its N-glycosylation at two arginine residues. Chimeric proteins consisting of PTTG1IP fused with various cargo proteins, and separated by self-cleaving sequences (to promote cargo release), were shown to enable highly efficient functional delivery of Cre protein to recipient cell cultures and mouse xenograft tumors, and delivery of Cas9-sgRNA complexes to recipient reporter cells. The favorable membrane topology of PTTG1IP enabled facile engineering of further variants with improved properties, highlighting its versatility and potential as a platform for EV-based therapeutics.

Published in Extracellular Vesicle

ISSN: 2773-0417 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General); Medicine: Medicine (General)
Website: https://www.sciencedirect.com/journal/extracellular-vesicle

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