Scientific publications

Efficient and Safe Therapeutic Use of Paired Cas9-Nickases for Primary Hyperoxaluria Type 1

Jan 16, 2024 | Magazine: EMBO Molecular Medicine

Laura Torella 1, Julia Klermund # 2 3, Martin Bilbao-Arribas # 1 4, Ibon Tamayo 4 5, Geoffroy Andrieux 6 7, Kay O Chmielewski 2 3, Africa Vales 1, Cristina Olagüe 1, Daniel Moreno-Luqui 1, Ivan Raimondi 1, Amaya Abad 1, Julen Torrens-Baile 1, Eduardo Salido 8, Maite Huarte 1, Mikel Hernaez 4 5, Melanie Boerries 6 7 9 10, Toni Cathomen # 11 12 13, Nerea Zabaleta # 14, Gloria Gonzalez-Aseguinolaza # 15


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.

CITA DEL ARTÍCULO EMBO Mol Med. 2024 Jan;16(1):112-131. doi: 10.1038/s44321-023-00008-8. Epub 2024 Jan 5.

Our authors

África Vales Aranguren
Ibon Tamayo Uría
Research Collaborator Bioinformatics Platform
Cristina Olagüe Micheltorena
Daniel Moreno Luqui
Julen Torrens Baile
Bioinformatics Research Technician Hepatic Diseases Gene Therapy Research Group
Dr. Mikel
Researcher | Principal Investigator Computational Biology Program