In vivo CRISPR base editing of PCSK9 durably lowers cholesterol in primates

被引：531

作者：

Musunuru, Kiran ^{[1
,2
,3
]}

Chadwick, Alexandra C. ^{[4
]}

Mizoguchi, Taiji ^{[4
]}

Garcia, Sara P. ^{[4
]}

DeNizio, Jamie E. ^{[4
]}

Reiss, Caroline W. ^{[4
]}

Wang, Kui ^{[4
]}

Iyer, Sowmya ^{[4
]}

Dutta, Chaitali ^{[4
]}

Clendaniel, Victoria ^{[4
]}

Amaonye, Michael ^{[4
]}

Beach, Aaron ^{[4
]}

Berth, Kathleen ^{[4
]}

Biswas, Souvik ^{[4
]}

Braun, Maurine C. ^{[4
]}

Chen, Huei-Mei ^{[4
]}

Colace, Thomas, V ^{[4
]}

Ganey, John D. ^{[4
]}

Gangopadhyay, Soumyashree A. ^{[4
]}

Garrity, Ryan ^{[4
]}

Kasiewicz, Lisa N. ^{[4
]}

Lavoie, Jennifer ^{[4
]}

Madsen, James A. ^{[4
]}

Matsumoto, Yuri ^{[4
]}

Mazzola, Anne Marie ^{[4
]}

Nasrullah, Yusuf S. ^{[4
]}

Nneji, Joseph ^{[4
]}

Ren, Huilan ^{[4
]}

Sanjeev, Athul ^{[4
]}

Shay, Madeleine ^{[4
]}

Stahley, Mary R. ^{[4
]}

Fan, Steven H. Y. ^{[5
]}

Tam, Ying K. ^{[5
]}

Gaudelli, Nicole M. ^{[6
]}

Ciaramella, Giuseppe ^{[6
]}

Stolz, Leslie E. ^{[4
]}

Malyala, Padma ^{[4
]}

Cheng, Christopher J. ^{[4
]}

Rajeev, Kallanthottathil G. ^{[4
]}

Rohde, Ellen ^{[4
]}

Bellinger, Andrew M. ^{[4
]}

Kathiresan, Sekar ^{[4
]}

机构：

[1] Univ Penn, Perelman Sch Med, Cardiovasc Inst, Philadelphia, PA 19104 USA

[2] Univ Penn, Dept Med, Perelman Sch Med, Div Cardiovasc Med, Philadelphia, PA 19104 USA

[3] Univ Penn, Perelman Sch Med, Dept Genet, Philadelphia, PA 19104 USA

[4] Verve Therapeut, Cambridge, MA 02139 USA

[5] Acuitas Therapeut, Vancouver, BC, Canada

[6] Beam Therapeut, Cambridge, MA USA

来源：

NATURE | 2021年 / 593卷 / 7859期

关键词：

GENOMIC DNA; ENDONUCLEASE; MUTATIONS; DISEASE; LDL;

D O I：

10.1038/s41586-021-03534-y

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Gene-editing technologies, which include the CRISPR-Cas nucleases(1-3) and CRISPR base editors(4,5), have the potential to permanently modify disease-causing genes in patients(6). The demonstration of durable editing in target organs of nonhuman primates is a key step before in vivo administration of gene editors to patients in clinical trials. Here we demonstrate that CRISPR base editors that are delivered in vivo using lipid nanoparticles can efficiently and precisely modify disease-related genes in living cynomolgus monkeys (Macaca fascicularis). We observed a near-complete knockdown of PCSK9 in the liver after a single infusion of lipid nanoparticles, with concomitant reductions in blood levels of PCSK9 and low-density lipoprotein cholesterol of approximately 90% and about 60%, respectively; all of these changes remained stable for at least 8 months after a single-dose treatment. In addition to supporting a 'once-and-done' approach to the reduction of low-density lipoprotein cholesterol and the treatment of atherosclerotic cardiovascular disease (the leading cause of death worldwide(7)), our results provide a proof-of-concept for how CRISPR base editors can be productively applied to make precise single-nucleotide changes in therapeutic target genes in the liver, and potentially in other organs.

引用

页码：429 / +

页数：22

共 50 条

[1] Mutations in PCSK9 cause autosomal dominant hypercholesterolemia [J].

Abifadel, M ;

Varret, M ;

Rabès, JP ;

Allard, D ;

Ouguerram, K ;

Devillers, M ;

Cruaud, C ;

Benjannet, S ;

Wickham, L ;

Erlich, D ;

Derré, A ;

Villéger, L ;

Farnier, M ;

Beucler, I ;

Bruckert, E ;

Chambaz, J ;

Chanu, B ;

Lecerf, JM ;

Luc, G ;

Moulin, P ;

Weissenbach, J ;

Prat, A ;

Krempf, M ;

Junien, C ;

Seidah, NG ;

Boileau, C .

NATURE GENETICS, 2003, 34 (02) :154-156

[2] Search-and-replace genome editing without double-strand breaks or donor DNA [J].

Anzalone, Andrew V. ;

Randolph, Peyton B. ;

Davis, Jessie R. ;

Sousa, Alexander A. ;

Koblan, Luke W. ;

Levy, Jonathan M. ;

Chen, Peter J. ;

Wilson, Christopher ;

Newby, Gregory A. ;

Raguram, Aditya ;

Liu, David R. .

NATURE, 2019, 576 (7785) :149-+

[3] Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases [J].

Bae, Sangsu ;

Park, Jeongbin ;

Kim, Jin-Soo .

BIOINFORMATICS, 2014, 30 (10) :1473-1475

[4] Trimmomatic: a flexible trimmer for Illumina sequence data [J].

Bolger, Anthony M. ;

Lohse, Marc ;

Usadel, Bjoern .

BIOINFORMATICS, 2014, 30 (15) :2114-2120

[5] Low Density Lipoprotein Cholesterol-Lowering Strategies and Population Health Time to Move to a Cumulative Exposure Model [J].

Brandts, Julia ;

Ray, Kausik K. .

CIRCULATION, 2020, 141 (11) :873-876

[6] In Vivo Base Editing of PCSK9 (Proprotein Convertase Subtilisin/Kexin Type 9) as a Therapeutic Alternative to Genome Editing [J].

Chadwick, Alexandra C. ;

Wang, Xiao ;

Musunuru, Kiran .

ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2017, 37 (09) :1741-+

[7] Full Coverage for Preventive Medications after Myocardial Infarction [J].

Choudhry, Niteesh K. ;

Avorn, Jerry ;

Glynn, Robert J. ;

Antman, Elliott M. ;

Schneeweiss, Sebastian ;

Toscano, Michele ;

Reisman, Lonny ;

Fernandes, Joaquim ;

Spettell, Claire ;

Lee, Joy L. ;

Levin, Raisa ;

Brennan, Troyen ;

Shrank, William H. .

NEW ENGLAND JOURNAL OF MEDICINE, 2011, 365 (22) :2088-2097

[8] CRISPResso2 provides accurate and rapid genome editing sequence analysis [J].

Clement, Kendell ;

Rees, Holly ;

Canver, Matthew C. ;

Gehrke, Jason M. ;

Farouni, Rick ;

Hsu, Jonathan Y. ;

Cole, Mitchel A. ;

Liu, David R. ;

Joung, J. Keith ;

Bauer, Daniel E. ;

Pinello, Luca .

NATURE BIOTECHNOLOGY, 2019, 37 (03) :224-226

[9] Sequence variations in PCSK9, low LDL, and protection against coronary heart disease [J].

Cohen, JC ;

Boerwinkle, E ;

Mosley, TH ;

Hobbs, HH .

NEW ENGLAND JOURNAL OF MEDICINE, 2006, 354 (12) :1264-1272

[10] CRISPOR: intuitive guide selection for CRISPR/Cas9 genome editing experiments and screens [J].

Concordet, Jean-Paul ;

Haeussler, Maximilian .

NUCLEIC ACIDS RESEARCH, 2018, 46 (W1) :W242-W245

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