Adoptive T cell therapy (ACT) is highly effective in the treatment of hematologic malignancies, but shows limited success in solid tumors. Inactivation of T cells in the tumor milieu is a major hurdle to a wider application of ACT. Cytotoxicity is the most relevant activity for tumor eradication. Here, we document that cytotoxic T cells (CTL) in lactic acidosis exhibited strongly reduced tumor cell killing, which could be compensated partly by increasing the CTL to tumor cell ratio. Lactic acid intervened at multiple steps of the killing process. Lactic acid repressed the number of CTL that performed lytic granule exocytosis (degranulation) in tumor cell co-culture, and, additionally impaired the quality of the response, as judged by the reduced intensity of degranulation and lower secretion of cytotoxins (perforin, granzyme B, granzyme A). CTL in lactic acid switched to a low bioenergetic profile with an inability to metabolize glucose efficiently. They responded to anti-CD3 stimulation poorly with less extracellular acidification rate (ECAR). This might explain their repressed granule exocytosis activity. Using live cell imaging, we show that CTL in lactic acid have reduced motility, resulting in lower field coverage. Many CTL in lactic acidosis did not make contact with tumor cells; however, those which made contact, adhered to the tumor cell much longer than a CTL in normal medium. Reduced motility together with prolonged contact duration hinders serial killing, a defining feature of killing potency, but also locally confines cytotoxic activity, which helps to reduce the risk of collateral organ damage. These activities define lactic acid as a major signaling molecule able to orchestrate the spatial distribution of CTL inside inflamed tissue, such as cancer, as well as moderating their functional response. Lactic acid intervention and strategies to improve T cell metabolic fitness hold promise to improve the clinical efficacy of T cell–based cancer immunotherapy.
Background Chronic viral infections caused by highly contagious human papillomaviruses (HPVs) from the alpha genus are a substantial risk factor for tumour diseases. Objectives The goal of this study was to compare the HPV infection pattern with histology in a patient group of immunocompromised HIV+ and non‐immunocompromised patients with anal intraepithelial neoplasia. Materials and Methods Tissue samples (n = 210) from the anogenital area of 121 patients underwent retrospective histological and molecular examination for HPV DNA prevalence by chip analysis. The study was part of a cancer screening from the Dermatology Department of the LMU Munich, Germany. All data were collected and processed anonymously. Results HPV 6 or 11 are more abundant in tissue samples from histologically diagnosed condylomata acuminata (47.7%) compared to grade 1, 2, and 3 intraepithelial neoplasias (IN 1‐3). Detection of high‐risk (hr) alpha‐HPV DNA was significantly higher in tissue samples from IN 3 (67.5%) compared to IN 1 and 2 (12.9%), and compared to condylomata acuminata (29.5%). No HPV types were detected in histologically unremarkable tissue samples. There was a significant association between the prevalence of HPV 16 and the classifications IN 1 to IN 3 (χ2 (2) = 13.62, P = 0.001). We identified a significant correlation between the prevalence of high‐risk and low‐risk (lr) HPV types and HIV, especially mixed infections of different HPV types correlated with high‐grade IN. Based on the present data, we suggest the risk of carcinoma in HIV+/− patients (RICH) score and test it in the 121 patients. Conclusions hr alpha‐HPVs, mainly HPV 16, are associated with increased oncogenic potential of premalignant lesions (IN 1‐3), especially in HIV+ patients. Based on the combination of HIV/HPV‐testing and histological analysis, we identified correlations that could potentially forecast the risk of malignant transformation and summarized them in the form of RICH score.
Introduction: The use of cellular immunotherapies has led to impressive complete and durable clinical responses in patients with certain types of hematological cancers. However, positive clinical results in solid tumor indications are still rare and many patients are in urgent need of alternative treatment options for several different indications. It has become clear that expression of inhibitory immune checkpoint molecules as well as harsh metabolic conditions in the tumor microenvironment (TME) are responsible for lack of activity of T cell immunotherapies in several settings, especially solid tumors. Here additional strategies are necessary to efficiently employ cellular immunotherapies. With the aim to further enhance the clinical efficacy of TCR-based immunotherapies under immunosuppressive conditions found in tumors, we analyzed the ability of PD1-41BB, a chimeric co-stimulatory receptor, to reverse the natural inhibitory PD-1/PD-L1 interaction into a supporting co-stimulatory signal in TCR-modified T cells encountering tumor cells. Methods: We evaluated the ability of the chimeric co-stimulatory receptor PD1-41BB to improve activity of TCR-modified T cells using 2-dimensional or 3-dimensional in vitro assays that model different immunosuppressive conditions found in tumors. Results: We demonstrate that chronic stimulation as well as several immunosuppressive factors of the TME, such as tumor cell expression of inhibitory immune checkpoint molecules or glucose restriction, impede the ability of TCR-transduced T cells to produce inflammatory cytokines and to efficiently lyse tumor cells. By using a chimeric co-stimulatory receptor consisting of the extracellular part of PD-1 and the co-stimulatory domain of 4-1BB we reversed the naturally occurring inhibitory PD-1/PD-L1 interaction to provide a co-stimulatory signal for improved T cell activity under immunosuppressive conditions or chronic stimulation. Addition of the chimeric co-stimulatory receptor PD1-41BB to TCR-modified T cells led to enhanced release of Interferon-γ, increased tumor cell killing, T cell proliferation and persistence in these T cell-tumor cell models. Conclusions: These preclinical studies support our approach to enhance the clinical efficacy of TCR-T therapies in PD-L1-positive malignancies by reversing naturally occurring inhibitory signals enabling counteraction of checkpoint-mediated dysfunction and metabolic insufficiency. The chimeric co-stimulatory PD1-41BB receptor has the potential to further enhance the clinical efficacy of TCR-modified T cells in patients with PD-L1-positive malignancies. Further preclinical in vitro and in vivo studies are ongoing to investigate the safety and efficacy of PD1-41BB in combination with multiple TCR candidates to explore its feasibility for the treatment of various cancers. Citation Format: Nadja Sailer, Melanie Salvermoser, Maria Gerget, Sarah Thome, Angelika J. Fischbeck, Svenja Ruehland, Luis F. Olguín-Contreras, Maja Buerdek, Christian Ellinger, Elfriede Noessner, Dolores J. Schendel, Patrik Kehler. The chimeric co-stimulatory receptor PD1-41BB enhances the function of T cell receptor (TCR)-modified T cells targeting solid tumors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3231.
Tumor spheroids, organoids, and cell culture models demand defined cell-cell and cell-matrix interactions, ideally with specific cell attachment on adhesion sites for multicellular spheroids or single cells. Our bioinert based micropatterning technology allows for the creation of adhesion spots while the surrounding passivated area is biologically inert, not interfering with the assay.We use this micro-patterning of adhesion ligands on highly passivated surfaces to generate homogenously distributed spheroids within a microfluidic channel system which can be perfused. Spheroids can either be directly generated by a simple injection of a large number of cells and a self-sorting process or by the addition of pre-formed spheroids, which will bind to the patterned adhesion sites. Perfusion of the spheroid chip enables the homogenous supply of all spheroids with nutrients and oxygen, and also the application of a defined shear stress, metabolite analysis together with toxicological screenings or even co-culture of multiple spheroid types. Applying a flow increases the overall spheroid growth, but condenses the cell aggregate, as well as influences the drug triggered protein expression response towards a more in vivo-like reaction.We also use this technique to generate arrays of homogenously distributed single cancer cells to evaluate cytotoxic T cell activity on a single cell level. By optical analysis combined with machine-learning based image processing of cell / spheroid arrays we obtained efficient label-free analysis of each cancer cell - T cell interaction over time. Citation Format: Elias Horn, Miriam Balles, Angelika J. Fischbeck, Jan Schwarz, Dane A. Thyssen, David W. Drell, Elfriede Noessner, Roman Zantl. Micropatterning supported method for cancer spheroid assays under flow and high throughput CAR-T cell potency assay with single cell resolution [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 314.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.