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Permanent alteration of PCSK9 in vivo genome editing

PCSK9 体内基因组编辑的永久改变

基本信息

批准号：
9307483
负责人：
Kiran Musunuru
金额：
$ 40.25万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-05-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9307483
关键词：
Adenoviruses Adult Antibodies Automobile Driving Blood Cardiovascular Diseases Cardiovascular system Cause of Death Cell Line Cholesterol Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complex Coronary heart disease Cytosine DNA Sequence Dependovirus Family Frameshift Mutation Future Genes Genomics Guide RNA Hepatocyte Human Human Genetics Human Genome Hybrids In Vitro Individual Injection of therapeutic agent Knock-out LDL Cholesterol Lipoproteins Light Liver Low Density Lipoprotein Receptor Mammalian Cell Mus Mutation Nonhomologous DNA End Joining Nonsense Mutation Nucleotides Patients Pharmaceutical Preparations Pharmacology Prevention Preventive therapy Production Proprotein Convertases Proteins Publishing Residual state Risk Risk Reduction Safety Sampling Site Streptococcus pyogenes Subtilisins System Technology Testing Thymine Time Transplantation Vaccination Work base cardiovascular risk factor deep sequencing disorder risk experience gain of function mutation genome editing genome-wide analysis humanized mouse hypercholesterolemia in vivo loss of function mutation mouse model new technology nuclease premature repaired success therapeutic target

项目摘要

PROJECT SUMMARY Proprotein convertase subtilisin/kexin type 9 (PCSK9) has emerged as a promising therapeutic target for the prevention of coronary heart disease (CHD). A gene specifically expressed in and secreted from the liver, and believed to function primarily as an antagonist to the low-density lipoprotein receptor (LDLR), PCSK9 was originally identified as the cause of autosomal dominant hypercholesterolemia in some families, with gain-of- function mutations in the gene driving highly elevated LDL cholesterol (LDL-C) levels and premature CHD. In subsequent studies, individuals with single loss-of-function mutations in PCSK9 were found to experience a significant reduction of both LDL-C levels (~30%–40%) as well as CHD risk (88%). Notably, even individuals with two loss-of-function mutations in PCSK9—resulting in ~80% reduction in LDL-C levels—appear to suffer no adverse clinical consequences. The ability to permanently alter the human genome has been made possible by the technology now commonly known as genome editing. Recently published clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated (Cas) systems use Streptococcus pyogenes Cas9 nuclease that is targeted to a genomic site by complexing with a synthetic guide RNA that hybridizes a 20-nucleotide DNA sequence (protospacer) immediately preceding an NGG motif (PAM, or protospacer-adjacent motif) recognized by Cas9. CRISPR-Cas9 generates a double-strand break (DSB) that is usually repaired by non-homologous end-joining (NHEJ), which is error-prone and conducive to frameshift mutations resulting in gene knock-out. A newer version of Cas9 termed a “base editor” selectively edits cytosine bases to thymine, without the need for DSBs, and thus may represent a safer means by which to introduce knock-out nonsense mutations. In light of the observed high efficiencies of CRISPR-Cas9 in mammalian cells in vitro, we seek to assess whether a one-time delivery of CRISPR-Cas9 can be used to permanently disrupt the human PCSK9 gene in vivo efficiently and safely and, if so, the optimal CRISPR-Cas9 system to use for this purpose. Success in completing this translational project will provide critical information on the feasibility of an in vivo genome- editing approach that could ultimately yield a one-shot, long-term therapy that permanently reduces blood LDL- C levels and thus serves as a “vaccination” against cardiovascular disease.

项目总结