Biology subjects

Kah, J.

Publications and source records attributed to Kah, J..

4 recordsLinked to original sources

bioRxiv2024

Accessing the specific capacity of TIL-derived CD8 T-cells to suppress tumor recurrence in resectable HBV-HCC patients

BackgroundHepatocellular carcinoma represents a significant global health challenge, affecting over a million patients annually, arising mainly from chronic liver diseases, with a majority being related to viral infections. However, despite the groundbreaking clinical results of immune checkpoint blockades and adoptive cell therapies, we still face non-responders accompanied by high rebound rates after resection. Considering that the main concern is to overcome a highly specialized immunosuppressive tumor microenvironment of the individual patient, the characterization of the particular tumor microenvironment and the source of the immune cells used in ACT is of immense importance. Approved ACT therapies mainly use modified peripheral blood cells from individuals. At the same time, tumor-infiltrating lymphocytes are underrepresented even if they have garnered interest due to their potential to target tumor-specific antigens more effectively. MethodsIn this study, we employed allogenic and autologous immune cell sources for expansion and stimulation, resulting in adoptive T-cell transfer experiments determining the effector cell differentiation and the related anti-tumor effects by the possible implementation of re-stimulation. ResultsWe determined a high success rate in expanding and stimulating tumor-infiltrating lymphocytes with consistent CD8 T-cell fractions from HCC patients. To showcase the effectiveness of stimulated T-cells from different sources, we generated cell lines derived from the margin and center of an HBV-induced HCC with a highly immune-suppressive TME. We found effective immune responses supported by cell death induction, ferroptosis, proptosis and apoptosis triggered by all sources of T-cells depending on the area of derived tumor cells. ConclusionEffector T-cell fractions derived from tumor-infiltrating lymphocytes present a viable source for cell-based immune therapy combined with immune checkpoint inhibitors in HCC patients, especially after resection, to suppress rebound strategies of the parental tumor on an individual level.

immunology↗

bioRxiv2024

Patient-derived HCC cells recapitulating the transcriptomic landscape of primary HCV-related liver cancer

BackgroundThe hepatocellular carcinoma is one of the leading causes of cancer-related mortality and is characterized by high heterogeneity and subsequently adaptation by developing resistance to current treatments. In this scenario the application of individualized models is crucial to understand the potential of approved therapies. Recently, we established a series of individual cell lines derived from patients who developed HCC on different entities, serving as a platform for individual approaches. In this study, we classified the LC4 cells derived from the center region of a HCC with underlying HIV-HCV co-infection, by using deep analysis on the pathway regulation level. MethodsWe employed DEG analysis, followed by pathway analysis to characterize the preservation level of the LC4 cells and the level of adoption. Next, we classify the model, by employing healthy donor samples, commonly used HCC cell lines and global RNAseq data sets. ResultsWe showed that the LC4 cells reflect significant characteristics of the parental region, including the replication of the immuno-suppressive and the proliferative milieu. The LC4 cells exhibit a metabolic reprogramming characterized by the downregulation of drug-metabolizing CYP enzymes compared to healthy individuals, indicating a transition to alternate metabolic pathways. Moreover, we identified common Biomarkers in the parental tissue, global datasets and the LC4 cells. ConclusionWe showed that the LC4 cell line is applicable as an individual model for pre-clinical testing of treatment regimens in HCC driven research.

cancer biology↗

bioRxiv2024

Ex-vivo expansion of patient-derived PBMCs preferentially results in effector memory T-cell proliferation with restored autologous efficiency in HBV-HCC

BackgroundT-cell-based therapies achieved milestones in targeting solid tumors, such as hepatocellular carcinoma, by leveraging the cytotoxic potential of effector memory T-cells. However, a key challenge lies in the ex-vivo expansion of functional memory T-cells while simultaneously preventing over-differentiation into senescent TEMRA cells. MethodsPBMC derived from 10 HCC patients and two healthy donors were used for expansion in small bioreactors equipped with a permeable membrane for 2 weeks. During expansion, surface marker composition, cytokine and chemokine production were observed. Enriched T-cell subsets from three patients with chronic HBV infection were analysed in detail. We extracted the enriched complement of one chronically infected patient to elucidate the therapeutic potential. ResultsWe successfully expanded the effector T-cell subsets for all investigated samples and consistently enriched the cell amount over time. Subsequently, we showed that the expanded patient-derived T-cells showed functionality against autologous liver cancer-derived cells by inducing receptor and protein-mediated cell death. ConclusionWe showed the consistent ex-vivo expansion of T-cell subsets of initial patient-derived PBMCs. The enriched subsets exhibit cytotoxic functionality and shift to cytolytic TEMRA cells in an immune evasion setting in the context of chronic HBV-infected patient-derived liver cancer cells.

immunology↗

bioRxiv2023

iMOUSE - Reforming the Strategy of Refinement and Reduction for indispensable laboratory animal-based studies in translational research

Considering the intricate nature of biological processes within organisms, it is undeniable that relying solely on in vitro-generated primary-cell-like cultures or organ-like products in preclinical and basic research is insufficient to replace animal-based studies fully. This limitation is particularly significant when considering the regulations enforced by legislative assemblies worldwide. The necessity of animal-based studies to approve chemicals and medications. In contradiction, European countries aim to banish animal-based studies. Therefore, we must understand the impact of the data refinement and experiment replacement strategy we will introduce here. This project aimed to revolutionize data acquisition in animal-based studies by transforming manual observation into a reliable digital process. Reliable digital data will be generated by having the potential to reduce human bias by simply reducing human interaction. Additionally, reducing human interaction will reduce the severity levels due to stress reduction, fulfilling the 3R principles. Therefore, the first goal was to develop and implement a scalable, stable, running, and remotely accessible camera-based monitor system (the iMouse solution). At the same time, the target was to develop a retrofit solution (DigiFrame) for existing home-cage systems, not interfering with the regular workflow in animal facilities. As a result, we developed a digital monitoring system, named iMouseTV platform based on existing open-source software, allowing users to observe, record, share, and review animal-based studies within the home cage anytime from anywhere, reducing the stress level for the animals. Our systems first Proof of concept ran for over two years at the LIV in Hamburg. We also investigated an effective way to reduce data generation by setting up specific zones for detecting the motion of choice (e.g., drinking, food intake). The data sets can be stored, shared, and reviewed by users and refined by algorithms aiming to recognize the dedicated motions of the animals automatically. The implementation of the ML algorithms allows the iMouse solution to recognize whether an individual mouse was drinking and for how long and store results in the annotated video file and graph format. However, the identification and continuous tracking of the species is still in progress. In conclusion, we established a scalable human-independent monitoring and recording system, which can be implemented into the existing structures of institutions and companies without changing handling processes, to monitor animals and observe them by getting reliable digital data. Moreover, it is fundamental for automatic recognition within animal-based studies based on Artificial Intelligence.

animal behavior and cognition↗