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Natural killer (NK) cells are potent cytotoxic innate lymphocytes that can be used for cancer immunotherapy. Since the balance of signals from activating and inhibitory receptors determines the activity of NK cells, their anti-tumor activity can be potentiated by overexpressing activating receptors or knocking out inhibitory receptors via genome engineering, such as chimeric antigen receptor (CAR) transgenesis and CRISPR-Cas9-mediated gene editing, respectively. Here, we report the development of a one-step strategy for CRISPR-Cas9-mediated gene knockout and CAR transgenesis in NK cells using retroviral particles. We generated NK cells expressing anti-epidermal growth factor receptor (EGFR)-CAR with simultaneous gene knockout using single transduction and evaluated the consequence of the genetic modifications and . Taken together, our results demonstrate that retroviral particle-mediated engineering provides a strategy readily applicable to simultaneous genetic modifications of NK cells for efficient immunotherapy.
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http://dx.doi.org/10.1016/j.omtm.2023.03.006 | DOI Listing |
EBioMedicine
August 2025
ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China; Shanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University, Shanghai, China; NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai, China; Instit
Advancements in gene therapy have achieved significant milestones in treating human diseases, offering renewed hope to patients with limited options. Key to this progress are vectors, which include both viral and non-viral methodologies that impact the success of gene therapy. Over the past two decades, three widely used viral vectors-lentiviruses (LV), adenoviruses (Ad), and adeno-associated viruses (AAV)-have enabled notable preclinical and clinical successes, including the approval of Luxturna for a genetic retinal disease and CAR-T therapies for blood cancers.
View Article and Find Full Text PDFViruses
May 2025
Lady Davis Institute, Jewish General Hospital, Montreal, QC H3T 1E2, Canada.
Lentiviral vectors (LVs) have become a fundamental tool in gene therapy due to their unique ability to transduce both dividing and non-dividing cells, transfer large genes of up to 10 kb, and facilitate stable, long-term expression of therapeutic genes into target cells. A key application of LVs is the ex vivo genetic modification of patient-derived cells, such as the production of CAR-T cells by transducing isolated T cells with LVs to express the CAR gene, enabling them to target and destroy cancer cells once infused back into the patient. However, these ex vivo gene therapy drugs are often dismally unaffordable due to the complex procedures involved, including cell isolation, genetic modification, and expansion, along with the significant risks associated with immune conditioning to ensure successful engraftment.
View Article and Find Full Text PDFFront Immunol
June 2025
Research and Development Immunotherapy, Miltenyi Biotec, Bergisch Gladbach, Germany.
γδ T cells represent a promising cell platform for adoptive cell therapy. Their natural anti-tumor reactivity and HLA-independent target cell recognition make them an attractive platform for allogeneic adoptive immunotherapy clinical interventions. Initial clinical trials exploring allogeneic γδ T-cell therapies have demonstrated encouraging safety profiles.
View Article and Find Full Text PDFCytotherapy
August 2025
Terumo BCT Asia Pte. Ltd., Singapore, Singapore. Electronic address:
Background Aims: Modifying a patient's own immune system to treat oncologic and autoimmune diseases using CAR-T therapy has become a major focus of research and clinical studies. While these therapies have immense capacity to treat and even cure diseases on a large scale, barriers such as complex manufacturing processes and long turnaround time impede widespread access. Traditional CAR-T manufacturing involves manual and open steps of activation and viral transduction before expansion, causing inefficiencies, inconsistencies, and extended timelines.
View Article and Find Full Text PDFNanomedicine
August 2025
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran 14115-111, Iran. Electronic address:
Genetic modification of immune cells remains a major challenge in immunotherapy. While viral and non-viral carriers exist, low gene transfer efficiency persists with non-viral methods. We present a peptide-based carrier (MiRGD) for gene delivery to diverse immune cells.
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