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Published on: August 2, 2018
In Vivo AAV-CRISPR/Cas9-Mediated Gene Editing Ameliorates Atherosclerosis in Familial Hypercholesterolemia
Huan Zhao1, Yan Li1, Lingjuan He1
1State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences (H.Z., Y.L., L.H., W.P., W.Y., Y.L., B.Z.).
Insights
Gene editing using CRISPR/Cas9 delivered by AAV in mice corrected the LDLR mutation, partially restoring protein expression and reducing atherosclerosis. This offers a potential therapy for familial hypercholesterolemia.
Area of Science:
- Cardiovascular Genetics
- Gene Therapy
- Molecular Medicine
Background:
- Familial hypercholesterolemia (FH) is caused by low-density lipoprotein (LDL) receptor (LDLR) mutations, leading to atherosclerosis and increased cardiovascular disease risk.
- The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system offers a promising avenue for gene editing to correct disease-causing mutations.
Purpose of the Study:
- To evaluate the efficacy of in vivo somatic cell gene editing via CRISPR/Cas9 delivered by adeno-associated virus (AAV) in a mouse model of FH.
- To determine if this approach could ameliorate atherosclerosis in mice with LDLR mutations.
Main Methods:
- A mouse model (Ldlr^null) with a nonsense point mutation in the LDLR gene was generated.
- CRISPR/Cas9 targeting the LDLR gene was delivered to hepatocytes using AAV in Ldlr^null mice.
- Gene correction, LDLR protein restoration, and atherosclerotic phenotypes were assessed.
Main Results:
- Ldlr^null mice on a high-fat diet exhibited severe atherosclerosis.
- AAV-CRISPR/Cas9 treatment partially corrected the LDLR mutation in hepatocytes and restored LDLR protein expression.
- Treated mice showed significant reductions in total cholesterol, triglycerides, and LDL cholesterol, with smaller aortic atherosclerotic plaques and less macrophage infiltration compared to controls.
Conclusions:
- In vivo CRISPR/Cas9-mediated LDLR gene correction using AAV effectively ameliorates atherosclerosis in a mouse model of FH.
- This approach shows potential as a therapeutic strategy for patients with familial hypercholesterolemia.
Background:
Mutations in low-density lipoprotein (LDL) receptor (LDLR) are one of the main causes of familial hypercholesterolemia, which induces atherosclerosis and has a high lifetime risk of cardiovascular disease. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system is an effective tool for gene editing to correct gene mutations and thus to ameliorate disease.
Methods:
The goal of this work was to determine whether in vivo somatic cell gene editing through the CRISPR/Cas9 system delivered by adeno-associated virus (AAV) could treat familial hypercholesterolemia caused by the Ldlr mutant in a mouse model. We generated a nonsense point mutation mouse line, Ldlr, based on a relevant familial hypercholesterolemia-related gene mutation. The AAV-CRISPR/Cas9 was designed to correct the point mutation in the Ldlr gene in hepatocytes and was delivered subcutaneously into Ldlr mice.
Results:
We found that homogeneous Ldlr mice (n=6) exhibited severe atherosclerotic phenotypes after a high-fat diet regimen and that the Ldlr mutation was corrected in a subset of hepatocytes after AAV-CRISPR/Cas9 treatment, with LDLR protein expression partially restored (n=6). Compared with the control groups (n=6 each group), the AAV-CRISPR/Cas9 with targeted single guide RNA group (n=6) had significant reductions in total cholesterol, total triglycerides, and LDL cholesterol in the serum, whereas the aorta had smaller atherosclerotic plaques and a lower degree of macrophage infiltration.
Conclusions:
Our work shows that in vivo AAV-CRISPR/Cas9-mediated Ldlr gene correction can partially rescue LDLR expression and effectively ameliorate atherosclerosis phenotypes in Ldlr mutants, providing a potential therapeutic approach for the treatment of patients with familial hypercholesterolemia.
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