Abstract 1504: Effective iPSC differentiation for generation of CAR T cells with canonical T cell phenotype and CAR T function

Induced pluripotent stem cell (iPSC)-derived T (iT) cells represent a groundbreaking frontier in adoptive cell therapies with engineered T cells, poised to overcome pivotal limitations associated with conventional manufacturing methods. iPSCs offer an off-the-shelf source of therapeutic T cells with the potential for infinite expansion and straightforward genetic manipulation to ensure hypo-immunogenicity and introduce specific therapeutic functions, such as antigen specificity through a chimeric antigen receptor (CAR). Importantly, genetic engineering of iPSC offers the benefit of generating fully modified clonal lines that are amenable to rigorous safety assessments. Critical to harnessing the potential of iT cells is the development of a robust and clinically compatible production process. Current protocols for genetic engineering as well as differentiation protocols designed to mirror human hematopoiesis and T cell development, vary in efficiency and often contain non-compliant components, thereby rendering them unsuitable for clinical implementation. This comprehensive review centers on the remarkable progress made over the last decade in generating functional engineered T cells from iPSCs. Emphasis is placed on alignment with good manufacturing practice (GMP) standards, scalability, safety measures and quality controls, which constitute the fundamental prerequisites for clinical application. In conclusion, the focus on iPSC as a source promises standardized, scalable, clinically relevant, and potentially safer production of engineered T cells. This groundbreaking approach holds the potential to extend hope to a broader spectrum of patients and diseases, leading in a new era in adoptive T cell therapy.

Section: In Vitro T Lymphoid Developmentmentioning

confidence: 99%

Section: Expression Of the Car Transgenesmentioning

confidence: 99%

Section: Considerations For Cgmp Differentiation Of Ipsc To It Cellsmentioning

confidence: 99%

See 1 more Smart Citation

Engineered T cells from induced pluripotent stem cells: from research towards clinical implementation

Netsrithong,

Garcia-Perez,

Themeli

2024

“…Beyond preclinical testing of immunotherapeutics, PDO can be employed to produce tumor cell specific T cells from induced pluripotent stem cells. This strategy may enable the production of allogeneic "off-the-shelf" CAR T cells circumventing the laborious and expensive generation of autologous CAR T cells (74). Moreover, large drug screens were successfully conducted implementing automated organoid seeding using automated microscopy or destructive viability assays as readouts for drug efficacy paving the way for applications within the highly regulated clinical setting (75,76).…”

Section: Patient-derived Organoidsmentioning

confidence: 99%

The challenge of making the right choice: patient avatars in the era of cancer immunotherapies

Kayser,

Brauer,

Susanne

et al. 2023

Immunotherapies are a key therapeutic strategy to fight cancer. Diverse approaches are used to activate tumor-directed immunity and to overcome tumor immune escape. The dynamic interplay between tumor cells and their tumor(immune)microenvironment (T(I)ME) poses a major challenge to create appropriate model systems. However, those model systems are needed to gain novel insights into tumor (immune) biology and a prerequisite to accurately develop and test immunotherapeutic approaches which can be successfully translated into clinical application. Several model systems have been established and advanced into so-called patient avatars to mimic the patient´s tumor biology. All models have their advantages but also disadvantages underscoring the necessity to pay attention in defining the rationale and requirements for which the patient avatar will be used. Here, we briefly outline the current state of tumor model systems used for tumor (immune)biological analysis as well as evaluation of immunotherapeutic agents. Finally, we provide a recommendation for further development to make patient avatars a complementary tool for testing and predicting immunotherapeutic strategies for personalization of tumor therapies.

“…However, the generation of helper T-iPS-T cells in feeder-free conditions has not been reported. Recent studies have demonstrated that 3D organoids provide a good environment to differentiate CD4 + T cells from hematopoeitc stem/progenitor cells and iPSC-derived endomesodermal progenitors (17)(18)(19)(20).…”

Section: Introductionmentioning

confidence: 99%

Lineage tracing of T cell differentiation from T-iPSC by 2D feeder-free culture and 3D organoid culture

Ishiguro,

Iriguchi,

Asano

et al. 2023

IntroductionT cells induced from induced pluripotent stem cells(iPSCs) derived from antigen-specific T cells (T-iPS-T cells) are an attractive tool for T cell immunotherapy. The induction of cytotoxic T-iPS-T cells is well established in feeder-free condition for the aim of off-the-shelf production, however, the induction of helper T-iPS-T cells remains challenging.MethodsWe analyzed T-iPS-T cells matured in 3D organoid culture at different steps in the culture process at the single-cell level. T-iPS-T cell datasets were merged with an available human thymocyte dataset based in single-cell RNA sequencing (scRNA-seq). Particularly, we searched for genes crucial for generation CD4+ T-iPS-T cells by comparing T-iPS-T cells established in 2D feeder-free or 3D organoid culture.ResultsThe scRNA-seq data indicated that T-iPS-T cells are similar to T cells transitioning to human thymocytes, with SELENOW, GIMAP4, 7, SATB1, SALMF1, IL7R, SYTL2, S100A11, STAT1, IFITM1, LZTFL1 and SOX4 identified as candidate genes for the 2D feeder-free induction of CD4+ T-iPS-T cells.DiscussionThis study provides single cell transcriptome datasets of iPS-T cells and leads to further analysis for CD4+ T cell generation from T-iPSCs.