In vivo CRISPR-Cas9 inhibition of hepatic LDH as treatment of primary hyperoxaluria
Rebeca Martinez-Turrillas 1 2 , Angel Martin-Mallo 3 2 , Saray Rodriguez-Diaz 1 2 , Natalia Zapata-Linares 1 , Paula Rodriguez-Marquez 3 2 , Patxi San Martin-Uriz 3 2 , Amaia Vilas-Zornoza 3 2 4 , María E Calleja-Cervantes 3 2 , Eduardo Salido 5 6 , Felipe Prosper 1 3 7 2 4 , Juan R Rodriguez-Madoz 1 3 2 4
Genome-editing strategies, especially CRISPR-Cas9 systems, have substantially increased the efficiency of innovative therapeutic approaches for monogenic diseases such as primary hyperoxalurias (PHs).
We have previously demonstrated that inhibition of glycolate oxidase using CRISPR-Cas9 systems represents a promising therapeutic option for PH type I (PH1).
Here, we extended our work evaluating the efficacy of liver-specific inhibition of lactate dehydrogenase (LDH), a key enzyme responsible for converting glyoxylate to oxalate; this strategy would not be limited to PH1, being applicable to other PH subtypes.
In this work, we demonstrate a liver-specific inhibition of LDH that resulted in a drastic reduction of LDH levels in the liver of PH1 and PH3 mice after a single-dose delivery of AAV8 vectors expressing the CRISPR-Cas9 system, resulting in reduced urine oxalate levels and kidney damage without signs of toxicity. Deep sequencing analysis revealed that this approach was safe and specific, with no off-targets detected in the liver of treated animals and no on-target/off-tissue events.
Altogether, our data provide evidence that in vivo genome editing using CRISPR-Cas9 systems would represent a valuable tool for improved therapeutic approaches for PH.