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2024
Xue N., Hong D., Zhang D., et al. Engineering IscB to develop highly efficient miniature editing tools in mammalian cells and embryos. Molecular Cell (2024).
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2024
Wang X., Wu X., Tan B., et al. Allogeneic CD19-targeted CAR-T therapy in severe myositis and systemic sclerosis achieved durable remission and reversed extensive fibrotic damages. Cell (2024).
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2024
Yang L., Huo Y., Wang M., et al. Engineering APOBEC3A deaminase for highly accurate and efficient base editing. Nature Chemical Biology (2024).
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2023
Zhang, N., Liu, X., Qin, J., et al. LIGHT/TNFSF14 promotes CAR-T cell trafficking and cytotoxicity through reversing immunosuppressive tumor microenvironment. Molecular Therapy (2023).
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2023
Chen, L., Hong, M., Luan, C., et al. Adenine transversion editors enable precise, efficient A·T-to-C•G base editing in mammalian cells and embryos. Nature Biotechnology (2023).
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2022
Chen, L., Zhu, B., Ru, G., et al. Re-engineering the adenine deaminase TadA-8e for efficient and specific CRISPR-based cytosine base editing. Nature Biotechnology (2022).
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2022
Chen, L., Zhang, S., Xue, N., et al. Engineering a precise adenine base editor with minimal bystander editing. Nature Chemical Biology (2022).
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2022
Zhang, J., Hu, Y., Yang, J., et al. Non-viral, specifically targeted CAR-T cells achieve high safety and efficacy in B-NHL. Nature (2022).
Recently, chimeric antigen receptor (CAR)-T cell therapy has shown great promise in
treating haematological malignancies1–7
. However, CAR-T cell therapy currently has
several limitations8–12. Here we successfully developed a two-in-one approach to
generate non-viral, gene-specifc targeted CAR-T cells through CRISPR–Cas9. Using
the optimized protocol, we demonstrated feasibility in a preclinical study by inserting
an anti-CD19 CAR cassette into the AAVS1 safe-harbour locus. Furthermore, an
innovative type of anti-CD19 CAR-T cell with PD1 integration was developed and
showed superior ability to eradicate tumour cells in xenograft models. In adoptive
therapy for relapsed/refractory aggressive B cell non-Hodgkin lymphoma
(ClinicalTrials.gov, NCT04213469), we observed a high rate (87.5%) of complete
remission and durable responses without serious adverse events in eight patients.
Notably, these enhanced CAR-T cells were efective even at a low infusion dose
and with a low percentage of CAR+
cells. Single-cell analysis showed that the
electroporation method resulted in a high percentage of memory T cells in infusion
products, and PD1 interference enhanced anti-tumour immune functions, further
validating the advantages of non-viral, PD1-integrated CAR-T cells. Collectively, our
results demonstrate the high safety and efcacy of non-viral, gene-specifc integrated
CAR-T cells, thus providing an innovative technology for CAR-T cell therapy.
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2022
Fu, B., Liao, J., Chen, S. et al. CRISPR–Cas9-mediated gene editing of the BCL11A enhancer for pediatric β0/β0 transfusion-dependent β-thalassemia. Nature Medicine (2022).
Gene editing to disrupt the GATA1-binding site at the +58 BCL11A erythroid enhancer could induce γ-globin expression, which is a promising therapeutic strategy to alleviate β-hemoglobinopathy caused by HBB gene mutation. In the present study, we report the preliminary results of an ongoing phase 1/2 trial (NCT04211480) evaluating safety and efficacy of gene editing therapy in children with blood transfusion-dependent β-thalassemia (TDT). We transplanted BCL11A enhancer-edited, autologous, hematopoietic stem and progenitor cells into two children, one carrying the β0/β0 genotype, classified as the most severe type of TDT. Primary endpoints included engraftment, overall survival and incidence of adverse events (AEs). Both patients were clinically well with multilineage engraftment, and all AEs to date were considered unrelated to gene editing and resolved after treatment. Secondary endpoints included achieving transfusion independence, editing rate in bone marrow cells and change in hemoglobin (Hb) concentration. Both patients achieved transfusion independence for >18 months after treatment, and their Hb increased from 8.2 and 10.8 g dl−1 at screening to 15.0 and 14.0 g dl−1 at the last visit, respectively, with 85.46% and 89.48% editing persistence in bone marrow cells. Exploratory analysis of single-cell transcriptome and indel patterns in edited peripheral blood mononuclear cells showed no notable side effects of the therapy.
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2020
Zhang, X., Zhu, B., Chen, L. et al. Dual base editor catalyzes both cytosine and adenine base conversions in human cells. Nature Biotechnology (2020).
Although base editors are useful tools for precise genome
editing, current base editors can only convert either adenines
or cytosines. We developed a dual adenine and cytosine base
editor (A&C-BEmax) by fusing both deaminases with a Cas9
nickase to achieve C-to-T and A-to-G conversions at the same
target site. Compared to single base editors, A&C-BEmax’s
activity on adenines is slightly reduced, whereas activity on
cytosines is higher and RNA off-target activity is substan-
tially decreased.
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2020
Zhang X., Chen L., Zhu B., et al. Increasing the efficiency and targeting range of cytidine base editors through fusion of a single-stranded DNA-binding protein domain. Nature Cell Biology (2020).
Cytidine base editors are powerful genetic tools that catalyse cytidine to thymidine conversion at specific genomic loci,
and further improvement of the editing range and efficiency is critical for their broader applications. Through insertion of a
non-sequence-specific single-stranded DNA-binding domain from Rad51 protein between Cas9 nickase and the deaminases,
serial hyper cytidine base editors were generated with substantially increased activity and an expanded editing window
towards the protospacer adjacent motif in both cell lines and mouse embryos. Additionally, hyeA3A-BE4max selectively cata-
lysed cytidine conversion in TC motifs with a broader editing range and much higher activity (up to 257-fold) compared with
eA3A-BE4max. Moreover, hyeA3A-BE4max specifically generated a C-to-T conversion without inducing bystander mutations in
the haemoglobin gamma gene promoter to mimic a naturally occurring genetic variant for amelioration of β-haemoglobinopathy,
suggesting the therapeutic potential of the improved base editors.
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2020
Zeng J, Wu Y, Ren C, Bauer DE, et al. Therapeutic base editing of human hematopoietic stem cells. Nature Medicine, 2020.
Base editing by nucleotide deaminases linked to programmable DNA-binding proteins represents a
promising approach to permanently remedy blood disorders, although its application in engrafting
hematopoietic stem cells (HSCs) remains unexplored. Here we purified A3A (N57Q)-BE3 protein
for ribonucleoprotein (RNP) electroporation of human peripheral blood (PB) mobilized CD34+
hematopoietic stem and progenitor cells (HSPCs). We observed frequent on-target cytosine base
