CRISPR/Cas9-mediated glycolate oxidase disruption is an efficacious and safe treatment for primary hyperoxaluria type I

Abstract

CRISPR/Cas9 technology offers novel approaches for the development of new therapies for many unmet clinical needs, including a significant number of inherited monogenic diseases. However, in vivo correction of disease-causing genes is still inefficient, especially for those diseases without selective advantage for corrected cells. We reasoned that substrate reduction therapies (SRT) targeting non-essential enzymes could provide an attractive alternative. Here we evaluate the therapeutic efficacy of an in vivo CRISPR/Cas9-mediated SRT to treat primary hyperoxaluria type I (PH1), a rare inborn dysfunction in glyoxylate metabolism that results in excessive hepatic oxalate production causing end-stage renal disease. A single systemic administration of an AAV8-CRISPR/Cas9 vector targeting glycolate oxidase, prevents oxalate overproduction and kidney damage, with no signs of toxicity in Agxt1−/− mice. Our results reveal that CRISPR/Cas9-mediated SRT represents a promising therapeutic option for PH1 that can be potentially applied to other metabolic diseases caused by the accumulation of toxic metabolites.

In vitroandIn vivoGenetic Disease Modelling via NHEJ-precise deletions using CRISPR/Cas9

“…The vectors and gRNAs used for the generation of a KO model in vivo were previously designed and individually characterized(Zabaleta et al 2018)…”

The Value of Mouse Models of Rare Diseases: A Spanish Experience

“…Deletion of the glycolate oxidase gene, inhibition of its enzymatic product or suppressing its expression with short-interfering RNA (siRNA) resulted in a substantial reversal of the hyperoxaluric phenotype ( Martin-Higueras et al, 2016 ), the latter is currently being evaluated in a clinical trial with encouraging preliminary results. Therapies based on in vivo CRISPR-Cas9 technology are also a potential strategy for curing PH1 by substrate reduction with the administration of AAV-mediated glycolate oxidase-targeted guide RNAs ( Zabaleta et al, 2018 ). We have generated a Grhpr knockout mouse for PH2 ( Knight et al, 2012 ), and both Agxt and Grhpr mutant mice have been used to test the potential of inhibiting hepatic lactate dehydrogenase with siRNA to treat PH ( Lai et al, 2018 ), which has moved to a clinical trial.…”

Patent Highlights June–July 2019

“…In this case the solution might be to inhibit glycolate oxidase; the increased glycolate can be renally excreted. Although hopes rest on siRNAs [24] and even CISPR technology [25], the search for small-molecule inhibitors of glycolate oxidase is also ongoing, as evidenced by the recent development of a highfuture science group www.future-science.com throughput assay [26]. A smaller-scale assay has been used by the inventors to screen newly synthesized compounds, among which 4- ( Numerous studies have demonstrated that depletion of cholesterol from plasma membranes interferes with lipid raft membrane microdomains, which preferentially associate with kinases involved in PI3K/Akt signal transduction [27].…”

“…Published: 25 As in other striated muscles, cardiac myofibrils are organized into sarcomeres, a network of contractile and structural proteins that regulate cardiac muscle function. Abnormalities in the cardiac sarcomere are the driving causes for hypertrophic cardiomyopathy and heart failure with preserved ejection fraction (formerly known as diastolic heart failure) [38].…”