A C1qTNF3 collagen domain fusion chaperones diverse secreted proteins and anti-Aβ scFvs: Applications for gene therapies

Brenda D. Moore; Yong Ran; Marshall S. Goodwin; Kavitha Komatineni; Karen N. McFarland; Kristy Dillon; Caleb Charles; Danny Ryu; Xuefei Liu; Stefan Prokop; Benoit I. Giasson; Todd E. Golde; Yona Levites

Molecular Therapy: Methods & Clinical Development (Dec 2023)

A C1qTNF3 collagen domain fusion chaperones diverse secreted proteins and anti-Aβ scFvs: Applications for gene therapies

Brenda D. Moore,
Yong Ran,
Marshall S. Goodwin,
Kavitha Komatineni,
Karen N. McFarland,
Kristy Dillon,
Caleb Charles,
Danny Ryu,
Xuefei Liu,
Stefan Prokop,
Benoit I. Giasson,
Todd E. Golde,
Yona Levites

Affiliations

Brenda D. Moore: Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA; Center for Neurodegenerative Disease, Emory University, School of Medicine, Atlanta, GA, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA; Corresponding author: Brenda D. Moore, Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA.
Yong Ran: Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA; Center for Neurodegenerative Disease, Emory University, School of Medicine, Atlanta, GA, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA
Marshall S. Goodwin: Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA
Kavitha Komatineni: Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA
Karen N. McFarland: Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; Department of Neurology, University of Florida College of Medicine, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA
Kristy Dillon: Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA
Caleb Charles: Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA
Danny Ryu: Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA; Center for Neurodegenerative Disease, Emory University, School of Medicine, Atlanta, GA, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA
Xuefei Liu: Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA; Center for Neurodegenerative Disease, Emory University, School of Medicine, Atlanta, GA, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA
Stefan Prokop: Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA; Department of Pathology, University of Florida, Gainesville, FL, USA
Benoit I. Giasson: Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA
Todd E. Golde: Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA; Center for Neurodegenerative Disease, Emory University, School of Medicine, Atlanta, GA, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA
Yona Levites: Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA; Center for Neurodegenerative Disease, Emory University, School of Medicine, Atlanta, GA, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL, USA; McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA; Corresponding author: Yona Levites, Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, USA.

Journal volume & issue: Vol. 31
p. 101146

Abstract

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Enhancing production of protein cargoes delivered by gene therapies can improve efficacy by reducing the amount of vector or simply increasing transgene expression levels. We explored the utility of a 126-amino acid collagen domain (CD) derived from the C1qTNF3 protein as a fusion partner to chaperone secreted proteins, extracellular “decoy receptor” domains, and single-chain variable fragments (scFvs). Fusions to the CD domain result in multimerization and enhanced levels of secretion of numerous fusion proteins while maintaining functionality. Efficient creation of bifunctional proteins using the CD domain is also demonstrated. Recombinant adeno-associated viral vector delivery of the CD with a signal peptide resulted in high-level expression with minimal biological impact as assessed by whole-brain transcriptomics. As a proof-of-concept in vivo study, we evaluated three different anti-amyloid Aβ scFvs (anti-Aβ scFvs), alone or expressed as CD fusions, following viral delivery to neonatal CRND8 mice. The CD fusion increased half-life, expression levels, and improved efficacy for amyloid lowering of a weaker binding anti-Aβ scFv. These studies validate the potential utility of this small CD as a fusion partner for secretory cargoes delivered by gene therapy and demonstrate that it is feasible to use this CD fusion to create biotherapeutic molecules with enhanced avidity or bifunctionality.

Published in Molecular Therapy: Methods & Clinical Development

ISSN: 2329-0501 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Cytology
Website: http://www.cell.com/molecular-therapy-family/methods/latest-content

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