Genome biology, Journal Year: 2023, Volume and Issue: 24(1)
Published: April 24, 2023
Multiple genetic modifications may be required to develop potent off-the-shelf chimeric antigen receptor (CAR) T cell therapies. Conventional CRISPR-Cas nucleases install sequence-specific DNA double-strand breaks (DSBs), enabling gene knock-out or targeted transgene knock-in. However, simultaneous DSBs provoke a high rate of genomic rearrangements which impede the safety edited cells.
Language: Английский
Signal Transduction and Targeted Therapy, Journal Year: 2023, Volume and Issue: 8(1)
Published: Jan. 16, 2023
Abstract Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene-editing technology is the ideal tool of future for treating diseases by permanently correcting deleterious base mutations or disrupting disease-causing genes with great precision and efficiency. A variety efficient Cas9 variants derivatives have been developed to cope complex genomic changes that occur during diseases. However, strategies effectively deliver CRISPR system diseased cells in vivo are currently lacking, nonviral vectors target recognition functions may be focus research. Pathological physiological resulting from disease onset expected serve as identifying factors targeted delivery targets gene editing. Diseases both varied complex, choice appropriate methods different important. Meanwhile, there still many potential challenges identified when targeting CRISPR/Cas9 treatment. This paper reviews current developments three aspects, namely, type, vector, characteristics. Additionally, this summarizes successful examples clinical trials finally describes possible problems associated applications.
Language: Английский
Citations
307
Science Advances, Journal Year: 2023, Volume and Issue: 9(7)
Published: Feb. 15, 2023
T cell engineering has changed the landscape of cancer immunotherapy. Chimeric antigen receptor cells have demonstrated a remarkable efficacy in treatment B malignancies hematology. However, their clinical impact on solid tumors been modest so far. expressing an engineered (TCR-T cells) represent promising therapeutic alternative. The target repertoire is not limited to membrane proteins, and intrinsic features TCRs such as high sensitivity near-to-physiological signaling may improve tumor detection killing while improving persistence. In this review, we present results obtained with TCR-T targeting different families. We detail methods that developed identify optimize TCR candidate. also discuss challenges therapies, including toxicity assessment resistance mechanisms. Last, share some perspectives highlight future directions field.
Language: Английский
Citations
207
New England Journal of Medicine, Journal Year: 2023, Volume and Issue: 389(10), P. 899 - 910
Published: June 14, 2023
Cytidine deamination that is guided by clustered regularly interspaced short palindromic repeats (CRISPR) can mediate a highly precise conversion of one nucleotide into another — specifically, cytosine to thymine without generating breaks in DNA. Thus, genes be base-edited and rendered inactive inducing translocations other chromosomal aberrations. The use this technique patients with relapsed childhood T-cell leukemia being investigated.
Language: Английский
Citations
164
Nature Reviews Clinical Oncology, Journal Year: 2023, Volume and Issue: 20(11), P. 739 - 754
Published: Aug. 16, 2023
Language: Английский
Citations
136
Circulation, Journal Year: 2022, Volume and Issue: 147(3), P. 242 - 253
Published: Oct. 31, 2022
VERVE-101 is an investigational in vivo CRISPR base-editing medicine designed to alter a single DNA base the PCSK9 gene, permanently turn off hepatic protein production, and thereby durably lower low-density lipoprotein cholesterol. We test efficacy, durability, tolerability, potential for germline editing of studies nonhuman primates murine F1 progeny study.Cynomolgus monkeys were given intravenous infusion vehicle control (n=10) or at dose 0.75 mg/kg (n=4) 1.5 (n=22) with subsequent follow-up up 476 days. Two assessed editing, including sequencing sperm samples from sexually mature male treated genotyping offspring female mice surrogate (VERVE-101mu).Liver biopsies 14 days after dosing noted mean 46% 70% mg/kg, respectively. This translated into reductions blood (proprotein convertase subtilisin/kexin type 9) 67% 83% cholesterol 49% 69% doses, respectively, as time-weighted average change baseline between day 28 dosing. Liver safety monitoring transient rise alanine aminotransferase aspartate concentrations that fully resolved by no accompanying total bilirubin. In subset necropsied 1 year dosing, findings related identified on macroscopic histopathologic assessment liver other organs. study assess primates, collected showed evidence editing. Among 436 saturating VERVE-101mu, edit was transmitted 0 animals.VERVE-101 well tolerated led durable effects These results have supported initiation first-in-human clinical trial patients heterozygous familial hypercholesterolemia atherosclerotic cardiovascular disease.
Language: Английский
Citations
121
Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)
Published: Jan. 13, 2023
CRISPR-Cas gene editing has revolutionized experimental molecular biology over the past decade and holds great promise for treatment of human genetic diseases. Here we review development CRISPR-Cas9/Cas12/Cas13 nucleases, DNA base editors, prime RNA focusing on assessment improvement their precision safety, pushing limit specificity efficiency. We summarize capabilities limitations each CRISPR tool from to editing, highlight opportunities future improvements applications in basic research, as well therapeutic clinical considerations use patients.
Language: Английский
Citations
104
Cell, Journal Year: 2023, Volume and Issue: 186(21), P. 4567 - 4582.e20
Published: Oct. 1, 2023
CRISPR-Cas9 genome editing has enabled advanced T cell therapies, but occasional loss of the targeted chromosome remains a safety concern. To investigate whether Cas9-induced is universal phenomenon and evaluate its clinical significance, we conducted systematic analysis in primary human cells. Arrayed pooled CRISPR screens revealed that was generalizable across resulted partial entire chromosome, including preclinical chimeric antigen receptor cells with persisted for weeks culture, implying potential to interfere use. A modified manufacturing process, employed our first-in-human trial Cas9-engineered (NCT03399448), reduced while largely preserving efficacy. Expression p53 correlated protection from observed this protocol, suggesting both mechanism strategy engineering mitigates genotoxicity clinic.
Language: Английский
Citations
93
Cell stem cell, Journal Year: 2022, Volume and Issue: 29(10), P. 1428 - 1444.e9
Published: Oct. 1, 2022
Long-range gene editing by homology-directed repair (HDR) in hematopoietic stem/progenitor cells (HSPCs) often relies on viral transduction with recombinant adeno-associated vector (AAV) for template delivery. Here, we uncover unexpected load and prolonged persistence of AAV genomes their fragments, which trigger sustained p53-mediated DNA damage response (DDR) upon recruiting the MRE11-RAD50-NBS1 (MRN) complex inverted terminal repeats (ITRs). Accrual cell-cycle-arrested HSPCs led to its frequent integration, predominantly form transcriptionally competent ITRs, at nuclease on- off-target sites. Optimized delivery integrase-defective lentiviral (IDLV) induced lower less persistent DDR, improving clonogenic capacity efficiency long-term repopulating HSPCs. Because insertions fragments are IDLV, choice mitigates adverse impact genotoxic burden HDR should facilitate clinical translation HSPC therapy.
Language: Английский
Citations
84
Cancer Discovery, Journal Year: 2023, Volume and Issue: 13(4), P. 829 - 843
Published: March 24, 2023
Abstract The success of chimeric antigen receptor (CAR) T cells targeting B-cell malignancies propelled the field synthetic immunology and raised hopes to treat solid tumors in a similar fashion. Antigen escape paucity tumor-restricted CAR targets are recognized challenges fulfilling this prospect. Recent advances cell engineering extend toolbox receptors available calibrate sensitivity combine create adapted tumor-sensing cells. Emerging strategies lower threshold for effective recognition, when needed, enable composite recognition hold great promise overcoming tumor heterogeneity curbing off-tumor toxicities. Significance: Improving clinical efficacy therapies will require that overcome heterogeneous low-abundance target expression while minimizing reactivity normal tissues. design logic gating poised beyond malignancies.
Language: Английский
Citations
69
Nature, Journal Year: 2024, Volume and Issue: 627(8003), P. 416 - 423
Published: Feb. 28, 2024
Abstract Permanent epigenetic silencing using programmable editors equipped with transcriptional repressors holds great promise for the treatment of human diseases 1–3 . However, to unlock its full therapeutic potential, an experimental confirmation durable after delivery transient in vivo is needed. To this end, here we targeted Pcsk9 , a gene expressed hepatocytes that involved cholesterol homeostasis. In vitro screening different editor designs indicated zinc-finger proteins were best-performing DNA-binding platform efficient mouse A single administration lipid nanoparticles loaded editors’ mRNAs almost halved circulating levels PCSK9 nearly one year mice. Notably, and accompanying repressive marks also persisted forced liver regeneration, further corroborating heritability newly installed state. Improvements construct design resulted development all-in-one configuration term evolved engineered repressor (EvoETR). This design, which characterized by high specificity profile, reduced mice efficiency comparable obtained through conventional editing, but without causing DNA breaks. Our study lays foundation therapeutics are based on silencing.
Language: Английский
Citations
68