Base editing of haematopoietic stem cells rescues sickle cell disease in mice

Newby, Gregory A.; Yen, Jonathan; Woodard, Kaitly J.; Mayuranathan, Thiyagaraj; Lazzarotto, Cícera R.; Li, Yichao; Sheppard-Tillman, Heather; Porter, Shaina N.; Yao, Yu; Mayberry, Kalin; Everette, Kelcee A.; Jang, Yoonjeong; Podracky, Christopher J.; Thaman, Elizabeth; Lechauve, Christophe; Sharma, Akshay; Henderson, Jordana M.; Richter, Michelle F.; Zhao, Kevin T.; Miller, Shannon M.; Wang, Tina; Koblan, Luke W.; McCaffrey, Anton P.; Tisdale, John F.; Kalfa, Theodosia A.; Pruett-Miller, Shondra M.; Tsai, Shengdar Q.; Weiss, Mitchell J.; Liu, David R.

doi:10.1038/s41586-021-03609-w

Cited by 203 publications

(166 citation statements)

References 52 publications

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“…In principle, cytosine base editors (CBEs) ( Komor et al., 2016 ; Nishida et al., 2016 ) and adenine base editors (ABEs) ( Gaudelli et al., 2017 ) can together correct the majority of known disease-causing single-nucleotide variants ( Anzalone et al., 2020 ; Rees and Liu, 2018 ). We and others have applied BEs to correct pathogenic point mutations and rescue disease phenotypes in mice and nonhuman primates ( Koblan et al., 2021 ; Levy et al., 2020 ; Musunuru et al., 2021 ; Newby and Liu, 2021 ; Newby et al., 2021 ; Rothgangl et al., 2021 ; Suh et al., 2021 ; Yeh et al., 2020 ), highlighting the potential of in vivo base editing as a therapeutic strategy.…”

Section: Introductionmentioning

confidence: 99%

“…An ideal method for delivering gene editing agents in vivo would directly deliver proteins or ribonucleoproteins (RNPs) instead of DNA. The short lifetime of RNPs in cells limits opportunities for off-target editing, as demonstrated by previous reports that delivering BE RNPs instead of BE-encoding DNA or mRNA leads to substantially reduced off-target editing, typically without sacrificing on-target editing efficiency ( Doman et al., 2020 ; Newby et al., 2021 ; Rees et al., 2017 ). While we and others have previously reported successful base editing in the mouse inner ear and retina following local administration of lipid-encapsulated BE RNPs ( Jang et al., 2021 ; Yeh et al., 2018 ), no generalizable strategy for delivering BE RNPs to multiple tissues and organs in vivo has been reported previously.…”

Section: Introductionmentioning

confidence: 99%

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Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

Banskota

Raguram

Suh

et al. 2022

Cell

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277

217

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

Banskota

Raguram

Suh

et al. 2022

Cell

Self Cite

277

217

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show abstract

“…Surprisingly, a more recent BE study showed that BE mRNA is superior to BE RNP. 44 These contradictory findings most likely reflect differences in the targeted locus and/or the editing proteins in question, which would benefit from a larger-scale, multi-locus comparison that factors in potential differences between distinct editing platforms such as CRISPR/Cas9, CRISPR BE, and ZFN.…”

Section: Discussionmentioning

confidence: 99%

Intracellular RNase activity dampens zinc finger nuclease-mediated gene editing in hematopoietic stem and progenitor cells

Peterson

Venkataraman

Reddy

et al. 2022

Molecular Therapy - Methods & Clinical Development

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Over the past decade, numerous gene-editing platforms which alter host DNA in a highly specific and targeted fashion have been described. Two notable examples are zinc finger nucleases (ZFNs), the first gene-editing platform to be tested in clinical trials, and more recently, CRISPR/Cas9. Although CRISPR/Cas9 approaches have become arguably the most popular platform in the field, the therapeutic advantages and disadvantages of each strategy are only beginning to emerge. We have established a nonhuman primate (NHP) model that serves as a strong predictor of successful gene therapy and gene-editing approaches in humans; our recent work shows that ZFN-edited hematopoietic stem and progenitor cells (HSPCs) engraft at lower levels than CRISPR/Cas9-edited cells. Here, we investigate the mechanisms underlying this difference. We show that optimized culture conditions, including defined serum-free media, augment engraftment of gene-edited NHP HSPCs in a mouse xenograft model. Furthermore, we identify intracellular RNases as major barriers for mRNA-encoded nucleases relative to preformed enzymatically active CRISPR/Cas9 ribonucleoprotein (RNP) complexes. We conclude that CRISPR/Cas9 RNP gene editing is more stable and efficient than ZFN mRNA-based delivery and identify co-delivered RNase inhibitors as a strategy to enhance the expression of gene-editing proteins from mRNA intermediates.

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“…Application of BE in congenital diseases caused by point mutations is now under pre-clinical investigation. In mouse sickle cell disease (SCD) model, Adenine base editor (ABE8e-NRCH) demonstrated the ability of ameliorating sickling morphological characteristic of reticulocytes, and the ability of converting pathogenic hemoglobin subunit beta (HBB) to benign HBB (59). Koblan et al (60) recently reported their research using ABE to correct mutation (c.1824 C>T, G608G) in Lamin A/C gene (LMNA) which causes Hutchinson-Gilford progeria syndrome (HGPS).…”

Section: Base Editor (Be)mentioning

confidence: 99%

Gene Therapy for Cardiovascular Disease: Basic Research and Clinical Prospects

Cao

Xuan

Zhang

et al. 2021

Front. Cardiovasc. Med.

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In recent years, the vital role of genetic factors in human diseases have been widely recognized by scholars with the deepening of life science research, accompanied by the rapid development of gene-editing technology. In early years, scientists used homologous recombination technology to establish gene knock-out and gene knock-in animal models, and then appeared the second-generation gene-editing technology zinc-finger nucleases (ZFNs) and transcription activator–like effector nucleases (TALENs) that relied on nucleic acid binding proteins and endonucleases and the third-generation gene-editing technology that functioned through protein–nucleic acids complexes—CRISPR/Cas9 system. This holds another promise for refractory diseases and genetic diseases. Cardiovascular disease (CVD) has always been the focus of clinical and basic research because of its high incidence and high disability rate, which seriously affects the long-term survival and quality of life of patients. Because some inherited cardiovascular diseases do not respond well to drug and surgical treatment, researchers are trying to use rapidly developing genetic techniques to develop initial attempts. However, significant obstacles to clinical application of gene therapy still exists, such as insufficient understanding of the nature of cardiovascular disease, limitations of genetic technology, or ethical concerns. This review mainly introduces the types and mechanisms of gene-editing techniques, ethical concerns of gene therapy, the application of gene therapy in atherosclerosis and inheritable cardiovascular diseases, in-stent restenosis, and delivering systems.

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Base editing of haematopoietic stem cells rescues sickle cell disease in mice

Cited by 203 publications

References 52 publications

Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

Intracellular RNase activity dampens zinc finger nuclease-mediated gene editing in hematopoietic stem and progenitor cells

Gene Therapy for Cardiovascular Disease: Basic Research and Clinical Prospects

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