evaluating-multimodal-functionality-chimeric-antigen-receptor-t-cells

Evaluating Multimodal Functionality of Chimeric Antigen Receptor T Cells at Single-Cell Resolution through Optofluidics Analysis

Adoptive T-cell therapy is a novel treatment paradigm in which autologous T cells are genetically modified to express a tumor-targeting chimeric antigen ...

Universitätsklinikum Würzburg

The microbiome is a complex host factor and key determinant of the outcome of antibody-based and cellular immunotherapy. Its postbiotics are a blend of soluble commensal byproducts that are released into the host environment and have been associated with the regulation of immune homeostasis, particularly through impacts on epigenetics and cell signaling. In this study, we show that the postbiotic pentanoate is metabolized to citrate within the TCA cycle via both the acetyl- and succinyl-CoA entry points, a feature uniquely enabled by the chemical structure of the C5 aliphatic chain. We identified ATP-citrate lyase as the crucial factor that redirects pentanoate-derived citrate from the succinyl-CoA route to the nucleus, thereby linking metabolic output and histone acetylation. This epigenetic-metabolic crosstalk mitigated T cell exhaustion and promoted naive-like differentiation in pentanoate-programmed chimeric antigen receptor (CAR) T cells. The predictive and therapeutic potential of pentanoate was corroborated in two independent patient cohorts and three syngeneic models of CAR T adoptive therapy. Our data demonstrate that postbiotics are integrated into mitochondrial metabolism and subsequently incorporated as epigenetic imprints. This bridge between microbial and mammalian interspecies communication can ultimately impact T cell differentiation and efficacy.

Metabolization of microbial postbiotic pentanoate drives anti-cancer CAR T cells.

The successful application of CAR-T cells in the treatment of hematologic malignancies has fundamentally changed cancer therapy. With increasing numbers of registered CAR-T cell clinical trials, efforts are being made to streamline and reduce the costs of CAR-T cell manufacturing while improving their safety. To date, all approved CAR-T cell products have relied on viral-based gene delivery and genomic integration methods. While viral vectors offer high transfection efficiencies, concerns regarding potential malignant transformation coupled with costly and time-consuming vector manufacturing are constant drivers in the search for cheaper, easier-to-use, safer, and more efficient alternatives. In this review, we examine different non-viral gene transfer methods as alternatives for CAR-T cell production, their advantages and disadvantages, and examples of their applications. Transposon-based gene transfer methods lead to stable but non-targeted gene integration, are easy to handle, and achieve high gene transfer rates. Programmable endonucleases allow targeted integration, reducing the potential risk of integration-mediated malignant transformation of CAR-T cells. Non-integrating CAR-encoding vectors avoid this risk completely and achieve only transient CAR expression. With these promising alternative techniques for gene transfer, all avenues are open to fully exploiting the potential of next-generation CAR-T cell therapy and applying it in a wide range of applications.

Current Non-Viral-Based Strategies to Manufacture CAR-T Cells.

Chimeric antigen receptor (CAR) T cell therapy has achieved remarkable success in treating haematologic malignancies. However, the rise in clinical use has highlighted substantial challenges related to T cell- and tumour-intrinsic mechanisms. Additionally, the tumour microenvironment can render these treatments dysfunctional. Extensive attempts in the field are optimizing the key elements of CAR T cell products for therapy, including antigen specificity and affinity, metabolic fitness, phenotypic stability and manufacturing. Recent efforts in transcriptomic and epigenetic profiling, as well as high-throughput functional screening methods, have identified new classes of targets, binders and mechanisms to be exploited. Advances in gene editing and delivery offer opportunities to translate those strategies into clinical trials. Here we discuss the multifaceted exploration of CAR T cell engineering approaches and emerging directions, highlighting the available strategies that can be built on to create the next generation of cellular therapies.

Article	Year
Metabolization of microbial postbiotic pentanoate drives anti-cancer CAR T cells. bioRxiv : the preprint server for biology\| PubMed ID: 39314273	2025
Current Non-Viral-Based Strategies to Manufacture CAR-T Cells. International journal of molecular sciences\| PubMed ID: 39769449	2024
Fine tuning towards the next generation of engineered T cells. Nature biomedical engineering\| PubMed ID: 41073806	2025

Patrick Ho

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