Efficient and safe therapeutic use of paired Cas9-nickases for primary hyperoxaluria type 1

Year: 2024

Type: article

Abstract: The therapeutic use of adeno-associated viral vector (AAV)-mediated gene disruption using CRISPR-Cas9 is limited by potential off-target modifications and the risk of uncontrolled integration of vector genomes into CRISPR-mediated double-strand breaks. To address these concerns, we explored the use of AAV-delivered paired Staphylococcus aureus nickases (D10ASaCas9) to target the Hao1 gene for the treatment of primary hyperoxaluria type 1 (PH1). Our study demonstrated effective Hao1 gene disruption, a significant decrease in glycolate oxidase expression, and a therapeutic effect in PH1 mice. The assessment of undesired genetic modifications through CIRCLE-seq and CAST-Seq analyses revealed neither off-target activity nor chromosomal translocations. Importantly, the use of paired-D10ASaCas9 resulted in a significant reduction in AAV integration at the target site compared to SaCas9 nuclease. In addition, our study highlights the limitations of current analytical tools in characterizing modifications introduced by paired D10ASaCas9, necessitating the development of a custom pipeline for more accurate characterization. These results describe a positive advance towards a safe and effective potential long-term treatment for PH1 patients.

Source: EMBO Molecular Medicine

DOI: 10.1038/s44321-023-00008-8

Authors: Laura Torella, Julia Klermund, Martin Bilbao-Arribas, Ibon Tamayo, Geoffroy Andrieux, Kay O Chmielewski, Africa Vales, Cristina Olagüe, Daniel Moreno-Luqui, Ivan Raimondi, Amaya Abad, Julen Torrens-Baile, Eduardo Salido, Maite Huarte, Mikel Hernaez, Melanie Boerries, Toni Cathomen, Nerea Zabaleta, Gloria Gonzalez-Aseguinolaza

Institutions: Centro de Investigación Médica Aplicada, Universidad de Navarra, University of Freiburg, University Medical Center Freiburg, Navarre Institute of Health Research, Universidad de La Laguna, Hospital Universitario de Canarias, German Cancer Research Center, Deutsches Konsortium für Translationale Krebsforschung, Massachusetts Eye and Ear Infirmary, Harvard University

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $6,890

Funders: Oxalosis and Hyperoxaluria Foundation, European Commission, Deutsche Forschungsgemeinschaft, Bundesministerium für Bildung und Forschung, Instituto de Salud Carlos III, Agencia Estatal de Investigación, Spanish National Plan for Scientific and Technical Research and Innovation

OpenAlex ID: W4390620182