Concurrent transposon engineering and CRISPR/Cas9 genome editing of primary CLL-1 chimeric antigen receptor–natural killer cells

Gurney, Mark E.; O’Reilly, Eimear; Corcoran, Sarah; Sarah, B; Krawczyk, Janusz; Otto, Neil; Hermanson, David; Childs, Richard W.; Szegezdi, Éva; O’Dwyer, Michael

doi:10.1016/j.jcyt.2022.07.008

Cited by 15 publications

(8 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nanoplasmids were combined to PB and TcBuster transposons to manufacture CAR-T and CAR-NK cells, respectively [ 82 , 156 ]. In the latter case, primary, donor-derived NK cells were genetically modified to express a CAR targeting the C-type lectin-like molecule-1 (CLL-1/ C-Type Lectin Domain Family 12 Member A, CLEC12A) against acute myeloid leukemia (AML) cell lines and primary AML blasts.…”

Section: Non-viral Strategy For Adoptive Immunotherapymentioning

confidence: 99%

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Marie,

Scherman

2024

Genes

View full text Add to dashboard Cite

Until very recently, the major use, for gene therapy, specifically of linear or circular DNA, such as plasmids, was as ancillary products for viral vectors’ production or as a genetic template for mRNA production. Thanks to targeted and more efficient physical or chemical delivery techniques and to the refinement of their structure, non-viral plasmid DNA are now under intensive consideration as pharmaceutical drugs. Plasmids traditionally carry an antibiotic resistance gene for providing the selection pressure necessary for maintenance in a bacterial host. Nearly a dozen different antibiotic-free gene vectors have now been developed and are currently assessed in preclinical assays and phase I/II clinical trials. Their reduced size leads to increased transfection efficiency and prolonged transgene expression. In addition, associating non-viral gene vectors and DNA transposons, which mediate transgene integration into the host genome, circumvents plasmid dilution in dividing eukaryotic cells which generate a loss of the therapeutic gene. Combining these novel molecular tools allowed a significantly higher yield of genetically engineered T and Natural Killer cells for adoptive immunotherapies due to a reduced cytotoxicity and increased transposition rate. This review describes the main progresses accomplished for safer, more efficient and cost-effective gene and cell therapies using non-viral approaches and antibiotic-free gene vectors.

show abstract

Section: Non-viral Strategy For Adoptive Immunotherapymentioning

confidence: 99%

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Marie,

Scherman

2024

Genes

View full text Add to dashboard Cite

show abstract

“…Gurney and colleagues applied a non-viral approach to primary CAR NK cell production combining the TcBuster DNA transposon system targeting a C-type lectin-like molecule-1 (CLL-1/C-Type Lectin Domain Family 12 Member A, CLEC12) with a GMP-grade Epstein-Barr virus-transformed lymphoblastoid feeder cell (EBC-LCL) for expansion. This approach knocked out a negative regulator of NK cell stimulation, cytokine-inducible SH-2-containing protein (CISH), using CRISPR/Cas9 to enhance the functionality of CLL-1 CAR NK cells without requiring IL-15 stimulation [135].…”

Section: Manipulations In Manufacturingmentioning

confidence: 99%

The Black Hole: CAR T Cell Therapy in AML

Atilla

Benabdellah

2023

Cancers

View full text Add to dashboard Cite

Despite exhaustive studies, researchers have made little progress in the field of adoptive cellular therapies for relapsed/refractory acute myeloid leukemia (AML), unlike the notable uptake for B cell malignancies. Various single antigen-targeting chimeric antigen receptor (CAR) T cell Phase I trials have been established worldwide and have recruited approximately 100 patients. The high heterogeneity at the genetic and molecular levels within and between AML patients resembles a black hole: a great gravitational field that sucks in everything. One must consider the fact that only around 30% of patients show a response; there are, however, consequential off-tumor effects. It is obvious that a new point of view is needed to achieve more promising results. This review first introduces the unique therapeutic challenges of not only CAR T cells but also other adoptive cellular therapies in AML. Next, recent single-cell sequencing data for AML to assess somatically acquired alterations at the DNA, epigenetic, RNA, and protein levels are discussed to give a perspective on cellular heterogeneity, intercellular hierarchies, and the cellular ecosystem. Finally, promising novel strategies are summarized, including more sophisticated next-generation CAR T, TCR-T, and CAR NK therapies; the approaches with which to tailor the microenvironment and target neoantigens; and allogeneic approaches.

show abstract

“…Similar to CAR-T cells, NK cells lack effective gene transfer strategies ( 88 ). Both viral and non-viral vectors have been used to genetically engineer CAR-NK cells ( 89 ). While the transduction efficiency of retroviral vectors is high, these vectors may cause insertional mutations, carcinogenesis, and other adverse effects ( 90 ).…”

Section: Combined Applications Of Car-nk Cells and Immune Checkpoint ...mentioning

confidence: 99%

Clinical application and prospect of immune checkpoint inhibitors for CAR-NK cell in tumor immunotherapy

Yang

Zhao²,

Sun³

et al. 2023

Front. Immunol.

View full text Add to dashboard Cite

Chimeric antigen receptor (CAR) engineering of natural killer (NK) cells is an attractive research field in tumor immunotherapy. While CAR is genetically engineered to express certain molecules, it retains the intrinsic ability to recognize tumor cells through its own receptors. Additionally, NK cells do not depend on T cell receptors for cytotoxic killing. CAR-NK cells exhibit some differences to CAR-T cells in terms of more precise killing, numerous cell sources, and increased effectiveness in solid tumors. However, some problems still exist with CAR-NK cell therapy, such as cytotoxicity, low transfection efficiency, and storage issues. Immune checkpoints inhibit immune cells from performing their normal killing function, and the clinical application of immune checkpoint inhibitors for cancer treatment has become a key therapeutic strategy. The application of CAR-T cells and immune checkpoint inhibitors is being evaluated in numerous ongoing basic research and clinical studies. Immune checkpoints may affect the function of CAR-NK cell therapy. In this review, we describe the combination of existing CAR-NK cell technology with immune checkpoint therapy and discuss the research of CAR-NK cell technology and future clinical treatments. We also summarize the progress of clinical trials of CAR-NK cells and immune checkpoint therapy.

show abstract

Concurrent transposon engineering and CRISPR/Cas9 genome editing of primary CLL-1 chimeric antigen receptor–natural killer cells

Cited by 15 publications

References 58 publications

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

The Black Hole: CAR T Cell Therapy in AML

Clinical application and prospect of immune checkpoint inhibitors for CAR-NK cell in tumor immunotherapy

Contact Info

Product

Resources

About