An in vivo Cell-Based Delivery Platform for Zinc Finger Artificial Transcription Factors in Pre-clinical Animal Models

Peter Deng; Julian A N M Halmai; Ulrika Beitnere; David Cameron; Michele L Martinez; Charles C Lee; Jennifer J Waldo; Krista Thongphanh; Anna Adhikari; Nycole Copping; Stela P Petkova; Ruth D Lee; Samantha Lock; Miranda Palomares; Henriette O'Geen; Jasmine Carter; Casiana E Gonzalez; Fiona K B Buchanan; Johnathan D Anderson; Fernando A Fierro; Jan A Nolta; Alice F Tarantal; Jill L Silverman; David J Segal; Kyle D Fink

doi:10.3389/fnmol.2021.789913

An in vivo Cell-Based Delivery Platform for Zinc Finger Artificial Transcription Factors in Pre-clinical Animal Models

Front Mol Neurosci. 2022 Jan 27:14:789913. doi: 10.3389/fnmol.2021.789913. eCollection 2021.

Authors

Peter Deng^{1

2

3

4}, Julian A N M Halmai^{1

2

4}, Ulrika Beitnere³, David Cameron^{1

2

4}, Michele L Martinez⁵, Charles C Lee⁵, Jennifer J Waldo^{1

2

4}, Krista Thongphanh^{1

2}, Anna Adhikari⁴, Nycole Copping⁴, Stela P Petkova⁴, Ruth D Lee⁴, Samantha Lock^{1

2

4}, Miranda Palomares³, Henriette O'Geen³, Jasmine Carter^{1

2

4}, Casiana E Gonzalez^{1

2

4}, Fiona K B Buchanan^{1

2

4}, Johnathan D Anderson⁶, Fernando A Fierro², Jan A Nolta², Alice F Tarantal⁵, Jill L Silverman⁴, David J Segal³, Kyle D Fink^{1

2

4}

Affiliations

¹ Department of Neurology, University of California Davis School of Medicine, Sacramento, CA, United States.
² Stem Cell Program and Gene Therapy Center, University of California, Davis, Sacramento, CA, United States.
³ Department of Biochemistry and Molecular Medicine, Genome Center, University of California, Davis, Davis, CA, United States.
⁴ Department of Psychiatry and Behavioral Sciences, MIND Institute, University of California Davis School of Medicine, Sacramento, CA, United States.
⁵ Departments of Pediatrics and Cell Biology and Human Anatomy, School of Medicine, Gene Therapy Center, and California National Primate Research Center, University of California, Davis, Davis, CA, United States.
⁶ Department of Otolaryngology, University of California, Davis, Davis, CA, United States.

Abstract

Zinc finger (ZF), transcription activator-like effectors (TALE), and CRISPR/Cas9 therapies to regulate gene expression are becoming viable strategies to treat genetic disorders, although effective in vivo delivery systems for these proteins remain a major translational hurdle. We describe the use of a mesenchymal stem/stromal cell (MSC)-based delivery system for the secretion of a ZF protein (ZF-MSC) in transgenic mouse models and young rhesus monkeys. Secreted ZF protein from mouse ZF-MSC was detectable within the hippocampus 1 week following intracranial or cisterna magna (CM) injection. Secreted ZF activated the imprinted paternal Ube3a in a transgenic reporter mouse and ameliorated motor deficits in a Ube3a deletion Angelman Syndrome (AS) mouse. Intrathecally administered autologous rhesus MSCs were well-tolerated for 3 weeks following administration and secreted ZF protein was detectable within the cerebrospinal fluid (CSF), midbrain, and spinal cord. This approach is less invasive when compared to direct intracranial injection which requires a surgical procedure.

Keywords: Angelman Syndrome (AS); artificial transcription factor (ATF); cell-based delivery; mesenchymal stem/stromal cell; zinc finger.

Copyright © 2022 Deng, Halmai, Beitnere, Cameron, Martinez, Lee, Waldo, Thongphanh, Adhikari, Copping, Petkova, Lee, Lock, Palomares, O’Geen, Carter, Gonzalez, Buchanan, Anderson, Fierro, Nolta, Tarantal, Silverman, Segal and Fink.

Abstract

Grants and funding