Adoptive cell therapy of prostate cancer using female mice-derived T cells that react with prostate antigens

BackgroundPeripheral blood natural killer (NK) cells are mature cytotoxic innate lymphocytes possessing an inherent capacity for tumor cell killing, thus making them attractive candidates for adoptive cell therapy. These NK cells are also amenable to CRISPR and chimeric antigen receptor (CAR) genomic engineering for enhanced functions. Moreover, NK cells possess an inherent capacity for off-the-shelf therapy since they are not known to cause graft-versus-host disease, unlike T cells. Presently, approved CAR cell therapy is custom-made from each patient‘s own T cells, a process that can limit patient pool, narrow therapeutic window, and contribute to product variability. In this study, we investigate whether peripheral blood NK cells from a selected donor can be edited, engineered, and expanded sufficiently for off-the-shelf use in a wide patient population.MethodsUsing the CRISPR/Cas9 system, we knocked out CISH expression in isolated peripheral blood NK cells from 3 healthy donors. Subsequently, we expanded edited NK cells by using IL-2 and sequential stimulations using NKSTIM, a modified K562 stimulatory cell line expressing membrane-bound form of IL-15 (mbIL-15) and 4-1BBL. IL-12 and IL-18 were added twice during expansion to drive memory-like NK cell differentiation. We transduced the expanded NK cells to express engineered CD19-targeted CAR and mbIL-15 during an interval between the first and second NKSTIM pulses. We assessed NK cell cytotoxicity against Nalm6 target cells by IncuCyte.ResultsIsolated peripheral blood NK cells from 3 healthy donors were successfully edited using CRISPR/Cas9, engineered to express high levels of CAR, extensively expanded using a series of NKSTIM pulses in the presence of IL-2, and differentiated into memory-like NK cells using IL-12 and IL-18. Interestingly, NK cells from the 3 donors exhibited distinct outcomes. NK cells from one donor reached a peak expansion limit of approximately 7-million-fold before undergoing contraction whereas NK cells from two donors continued to expand over the length of the study surpassing 100-million-fold expansion, without appearing to have reached a terminal expansion limit. At the end of the study, NK cells from one donor exceeded 1-billion-fold expansion and maintained 88% cytolytic activity compared to Nkarta’s standard process control in a 72-hour IncuCyte assay.ConclusionsIn this study, we demonstrate that healthy donor-derived peripheral blood NK cells are capable of expanding over billion-fold while maintaining potency. These results provide a rationale for the development of off-the-shelf CAR NK cell therapies using NK cells from donors selected to provide optimal product characteristics.Ethics ApprovalHuman samples were collected with written informed consent by an approved vendor.

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Advances in Adoptive Cell Therapy Using Induced Pluripotent Stem Cell-Derived T Cells

Frontiers in Immunology ◽

10.3389/fimmu.2021.759558 ◽

2021 ◽

Vol 12 ◽

Author(s):

Ratchapong Netsrithong ◽

Methichit Wattanapanitch

Keyword(s):

T Cells ◽

T Cell ◽

Cell Therapy ◽

Cell Biology ◽

Ex Vivo ◽

Adoptive Cell Therapy ◽

Basic Knowledge ◽

Car T Cells ◽

Car T ◽

Induced Pluripotent

Adoptive cell therapy (ACT) using chimeric antigen receptor (CAR) T cells holds impressive clinical outcomes especially in patients who are refractory to other kinds of therapy. However, many challenges hinder its clinical applications. For example, patients who undergo chemotherapy usually have an insufficient number of autologous T cells due to lymphopenia. Long-term ex vivo expansion can result in T cell exhaustion, which reduces the effector function. There is also a batch-to-batch variation during the manufacturing process, making it difficult to standardize and validate the cell products. In addition, the process is labor-intensive and costly. Generation of universal off-the-shelf CAR T cells, which can be broadly given to any patient, prepared in advance and ready to use, would be ideal and more cost-effective. Human induced pluripotent stem cells (iPSCs) provide a renewable source of cells that can be genetically engineered and differentiated into immune cells with enhanced anti-tumor cytotoxicity. This review describes basic knowledge of T cell biology, applications in ACT, the use of iPSCs as a new source of T cells and current differentiation strategies used to generate T cells as well as recent advances in genome engineering to produce next-generation off-the-shelf T cells with improved effector functions. We also discuss challenges in the field and future perspectives toward the final universal off-the-shelf immunotherapeutic products.

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Adoptive cell therapy for cancer using T-cells genetically engineered to express chimeric antigen receptors

Hematopoietic Cell Transplants ◽

10.1017/9781316335727.063 ◽

2017 ◽

pp. 591-601

Author(s):

Sarwish Rafiq ◽

Renier J. Brentjens

Keyword(s):

T Cells ◽

Cell Therapy ◽

Adoptive Cell Therapy ◽

Genetically Engineered ◽

Antigen Receptors ◽

Chimeric Antigen Receptors

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Adoptive cell therapy of tolerogenic dendritic cells as inducer of regulatory T cells in allergic rhinitis

International Forum of Allergy & Rhinology ◽

10.1002/alr.22217 ◽

2018 ◽

Vol 8 (11) ◽

pp. 1291-1299 ◽

Cited By ~ 4

Author(s):

Wei Sun ◽

Jia-Wei Wei ◽

Hang Li ◽

Fan-Qin Wei ◽

Jian Li ◽

...

Keyword(s):

T Cells ◽

Allergic Rhinitis ◽

Dendritic Cells ◽

Regulatory T Cells ◽

Cell Therapy ◽

Adoptive Cell Therapy ◽

Tolerogenic Dendritic Cells

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An Update on Adoptive T-Cell Therapy and Neoantigen Vaccines

American Society of Clinical Oncology Educational Book ◽

10.1200/edbk_238001 ◽

2019 ◽

pp. e70-e78 ◽

Cited By ~ 11

Author(s):

Patrick A. Ott ◽

Gianpietro Dotti ◽

Cassian Yee ◽

Stephanie L. Goff

Keyword(s):

T Cells ◽

T Cell ◽

Cell Therapy ◽

Adoptive Cell Therapy ◽

Adoptive T Cell Therapy ◽

Clinical Activity ◽

Single Chain ◽

T Cell Therapy ◽

Substantial Impact ◽

Mhc Molecules

Adoptive T-cell therapy using tumor-infiltrating lymphocytes (TILs) has demonstrated long-lasting antitumor activity in select patients with advanced melanoma. Cancer vaccines have been used for many decades and have shown some promise but overall relatively modest clinical activity across cancers. Technological advances in genome sequencing capabilities and T-cell engineering have had substantial impact on both adoptive cell therapy and the cancer vaccine field. The ability to identify neoantigens—a class of tumor antigens that is truly tumor specific and encoded by tumor mutations through rapid and relatively inexpensive next-generation sequencing—has already demonstrated the critical importance of these antigens as targets of antitumor-specific T-cell responses in the context of immune checkpoint blockade and other immunotherapies. Therapeutically targeting these antigens with either adoptive T-cell therapy or vaccine approaches has demonstrated early promise in the clinic in patients with advanced solid tumors. Chimeric antigen receptor (CAR) T cells, which are engineered by fusing an antigen-specific, single-chain antibody (scFv) with signaling molecules of the T-cell receptor (TCR)/CD3 complex creating an antibody-like structure on T cells that recognizes antigens independently of major histocompatibility complex (MHC) molecules, have demonstrated remarkable clinical activity in patients with advanced B-cell malignancies, leading to several approvals by the U.S. Food and Drug Administration (FDA).

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