Genotoxic effects of base and prime editing in human hematopoietic stem cells

Nature Biotechnology, Journal Year: 2023, Volume and Issue: 42(6), P. 877 - 891

Published: Sept. 7, 2023

Base and prime editors (BEs PEs) may provide more precise genetic engineering than nuclease-based approaches because they bypass the dependence on DNA double-strand breaks. However, little is known about their cellular responses genotoxicity. Here, we compared state-of-the-art BEs PEs Cas9 in human hematopoietic stem progenitor cells with respect to editing efficiency, cytotoxicity, transcriptomic changes on-target genome-wide induced detrimental transcriptional that reduced efficiency repopulation xenotransplants also generated breaks genotoxic byproducts, including deletions translocations, at a lower frequency Cas9. These effects were strongest for cytidine due suboptimal inhibition of base excision repair mitigated by tailoring delivery timing editor expression through optimized mRNA design. altered mutational landscape across genome increasing load relative proportions nucleotide variants. findings raise concerns genotoxicity warrant further investigation view clinical application.

Language: Английский

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A new era of cancer immunotherapy: combining revolutionary technologies for enhanced CAR-M therapy

Molecular Cancer, Journal Year: 2024, Volume and Issue: 23(1)

Published: June 1, 2024

Abstract Significant advancements have been made in the application of chimeric antigen receptor (CAR)-T treatment for blood cancers during previous ten years. However, its effectiveness treating solid tumors is still lacking, necessitating exploration alternative immunotherapies that can overcome significant challenges faced by current CAR-T cells. CAR-based immunotherapy against shows promise with emergence macrophages, which possess robust phagocytic abilities, antigen-presenting functions, and ability to modify tumor microenvironment stimulate adaptive responses. This paper presents a thorough examination latest progress CAR-M therapy, covering both basic scientific studies clinical trials. study examines primary obstacles hindering realization complete potential as well strategies be employed these hurdles. With revolutionary technologies like situ genetic modification, synthetic biology techniques, biomaterial-supported gene transfer, provide wider array resources manipulating tumor-associated we suggest combining advanced methods will result creation new era therapy demonstrates improved efficacy, safety, availability. Graphical

Language: Английский

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Improving access to gene therapy for rare diseases

Disease Models & Mechanisms, Journal Year: 2024, Volume and Issue: 17(6)

Published: April 19, 2024

ABSTRACT Effective gene therapy approaches have been developed for many rare diseases, including inborn errors of immunity and metabolism, haemoglobinopathies inherited blindness. Despite successful pre-clinical clinical results, these therapies are not widely available, primarily non-medical reasons. Lack commercial interest in ultra-rare costs development complex manufacturing processes required advanced medicinal products (ATMPs) some the main problems that restricting access. The complexities navigating regulatory environments different jurisdictions treatments affect small numbers patients is a problem unique to ATMPS diseases. In this Perspective, we outline challenges potential solutions that, hope, will improve access

Language: Английский

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Advanced gene nanocarriers/scaffolds in nonviral-mediated delivery system for tissue regeneration and repair

Journal of Nanobiotechnology, Journal Year: 2024, Volume and Issue: 22(1)

Published: June 26, 2024

Abstract Tissue regeneration technology has been rapidly developed and widely applied in tissue engineering repair. Compared with traditional approaches like surgical treatment, the rising gene therapy is able to have a durable effect on regeneration, such as impaired bone articular cartilage repair cancer-resected Gene can also facilitate production of situ therapeutic factors, thus minimizing diffusion or loss complexes enabling spatiotemporally controlled release products for regeneration. Among different delivery vectors supportive gene-activated matrices, advanced gene/drug nanocarriers attract exceptional attraction due their tunable physiochemical properties, well excellent adaptive performance bone, cartilage, blood vessel, nerve This paper reviews recent advances nonviral-mediated systems an emphasis important role

Language: Английский

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Exonic knockout and knockin gene editing in hematopoietic stem and progenitor cells rescues RAG1 immunodeficiency

Science Translational Medicine, Journal Year: 2024, Volume and Issue: 16(733)

Published: Feb. 7, 2024

Recombination activating genes ( RAGs ) are tightly regulated during lymphoid differentiation, and their mutations cause a spectrum of severe immunological disorders. Hematopoietic stem progenitor cell (HSPC) transplantation is the treatment choice but limited by donor availability toxicity. To overcome these issues, we developed gene editing strategies targeting corrective sequence into human RAG1 homology-directed repair (HDR) validated them tailored two-dimensional, three-dimensional, in vivo xenotransplant platforms to assess rescue expression function. Whereas integration intron 1 achieved suboptimal correction, in-frame insertion exon 2 drove physiologic activity, allowing disruption dominant-negative effects unrepaired hypomorphic alleles. Enhanced HDR-mediated enabled correction HSPCs from patients with T B differentiation blocks. Gene efficiency exceeded minimal proportion functional required immunodeficiency Rag1 –/– mice, supporting clinical translation HSPC for deficiency.

Language: Английский

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Enhancing pediatric access to cell and gene therapies

Nature Medicine, Journal Year: 2024, Volume and Issue: 30(7), P. 1836 - 1846

Published: June 17, 2024

Language: Английский

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Impact of CRISPR/HDR editing versus lentiviral transduction on long-term engraftment and clonal dynamics of HSPCs in rhesus macaques

Cell stem cell, Journal Year: 2024, Volume and Issue: 31(4), P. 455 - 466.e4

Published: March 19, 2024

Language: Английский

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Adaptation to ex vivo culture reduces human hematopoietic stem cell activity independently of the cell cycle

Blood, Journal Year: 2024, Volume and Issue: 144(7), P. 729 - 741

Published: May 28, 2024

Loss of long-term hematopoietic stem cell (LT-HSC) function ex vivo hampers the success clinical protocols that rely on culture. However, kinetics and mechanisms through which this occurs remain incompletely characterized. In study, time-resolved single-cell RNA sequencing, matched in functional analysis, use a reversible vitro system early G1 arrest, we defined sequence transcriptional events occur during first division human LT-HSCs. We demonstrated sharpest loss LT-HSC repopulation capacity happens on, between 6 24 hours culture, before LT-HSCs commit to cycle progression. During time window, adapt culture environment, limit global variability gene expression, transiently upregulate networks involved signaling stress responses. From hours, progression past contributes establishment differentiation programs contrary current assumptions, HSC is independent Finally, showed targeting adaptation by inhibiting activation JAK/STAT improves repopulating vivo. Collectively, our study controlling for example, via JAK inhibition, critically important improve therapy expansion protocols.

Language: Английский

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Creating superior lungs for transplantation with next-generation gene therapy during ex vivo lung perfusion

The Journal of Heart and Lung Transplantation, Journal Year: 2024, Volume and Issue: 43(5), P. 838 - 848

Published: Feb. 2, 2024

Engineering donor organs to better tolerate the harmful non-immunological and immunological responses inherently related solid organ transplantation would improve transplant outcomes. Our enhanced knowledge of ischemia-reperfusion injury, alloimmune pathological fibroproliferation after transplantation, advanced toolkit available for gene therapies, have brought this goal closer clinical reality. Ex vivo perfusion has evolved rapidly especially in field lung where clinicians routinely use ex (EVLP) confirm quality marginal lungs before enabling safe originally considered unusable. EVLP also be an attractive platform deliver as treatments could administered isolated thereby providing a window sophisticated engineering while minimizing off-target effects recipient. Here, we review status first-generation therapies that focus on inducing transgene expression target cells. We highlight recent advances next-generation enable editing epigenetic engineering, used permanently change genome induce widespread transcriptional modulation lung. In future vision, dedicated repair centers will not only increase pool, but create superior function longer

Language: Английский

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CRISPR-Cas9n-mediated ELANE promoter editing for gene therapy of severe congenital neutropenia

Molecular Therapy, Journal Year: 2024, Volume and Issue: 32(6), P. 1628 - 1642

Published: March 30, 2024

Language: Английский

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