Regular physical activity has been associated with healthy brain aging, reflected by beneficial effects on cognition and learning and memory. Nutritional supplements such as caffeine have been shown to act as cognitive enhancers and may possess neuroprotective properties. Interestingly, caffeine also improves athletic capabilities and is widely used by athletes because of its performance-enhancing effect, while information on potential additive beneficial effects of physical activity and caffeine on cognitive performance is scarce. In the present study, the effects of caffeine supplementation in combination with prolonged physical and cognitive stimulation in the form of the enriched environment (EE) housing for a duration of 4 months were analyzed. We demonstrate that caffeine supplementation together with prolonged environmental enrichment led to enhanced memory function, resulting in improved recognition and spatial working memory in behavioral paradigms such as the novel object recognition task or the Morris water maze in C57Bl6 wild-type mice. Mice housed under EE conditions showed increased gene expression levels of brain-derived neurotrophic factor (BDNF) in the hippocampus. The present findings underscore the potential impact of continuous physical activity in the prevention of age-related cognitive decline and may offer new options for combinatorial approaches.

Physiologically relevant in vitro tumor models are crucial in any research setting from drug development to individualized treatment predictions. Patient-derived 3D cell culture models (PD3D®) are validated cancer models which recapitulate the biology of the donor tissue from genotype to phenotype. A multitude of PD3D® replicates can be generated and these are suitable for high-throughput-screening. However, the influence of media conditions on specific drug sensitivity profiles widely remains elusive and thus the predictive value of screenings is obscured. Here we have investigated pathway activities in dependence of different media compositions and mapped out differences in response to targeted therapies and distinct growth media. We assessed the response of PD3D® models derived from colorectal carcinoma (CRC) patients towards two targeted drugs aiming at different levels of the EGFR/Ras/Raf/Mek/Erk axis in defined, serum-free culture media. One media composition contained growth factors in concentration prevalently used in published cell culture protocols, while the other composition is geared to better match physiological levels of relevant supplements. We observed a significant impact of media conditions on sensitivity towards targeted cancer drugs. However, these differences were as individual as the models and the sensitivity against the treatment. To uncover the underlying mechanisms in the observed discrepancy we used DigiWest - a targeted proteomics technology - to assess pathways activity in the different models. We applied a panel of over 250 (phospho-)antibodies against proteins in the MAPK/ERK/RAS, PI3K/AKT and mTOR pathways as well as cell cycle and proliferation. Using functional pathway analysis, we mapped out the difference in response to targeted therapies and distinct growth media. In conclusion, media composition has great impact on the response to targeted drugs in vitro and therefore needs to be harmonized to ensure relevant results. Combination of PD3D® models and DigiWest functional pathway analysis represents a powerful tool to identify the molecular mechanisms underlying differential drug response. Further development of this collaborative approach will lead to a better understanding of the drug resistance or sensitivity and potentially even identification of signatures related to them. Citation Format: Jürgen Loskutov, Gerrit Erdmann, Anja Arndt, Quirin Graf Adelmann, Przemyslaw Dudys, Marie Flechner, Madeleine Nadolny, Katja Osman, Ulrike Pfohl, Christoph Reinhard, Markus Templin, Katja Uhlig, Lena Wedeken, Christian Regenbrecht. Let’s get physiological! Impact of media conditions on drug response to targeted therapies in CRC organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 187.

Enhanced, physiologically relevant in-vitro tumor models significantly increase the success rate in new drug development, reduce the number of animal experiments and result finally in better therapies for cancer patients. Furthermore, they have the potential to offer new approaches to personalized oncology. In recent years, 3D cell cultures were established as relevant preclinical cancer models, suitable for high-throughput-screening. However, their capacity to model and/or measure physiological processes can still be enhanced. Colorectal carcinomas (CRC) are one of the most common malignant tumors. Due to its heterogeneity and high probability to metastasize, the disease is often incurable and novel, improved therapies are urgently needed. In this project, we developed a tumor organoid-on-chip platform (TumOC) by combining most recent technological advances: We integrated CRC patient-derived 3D cell cultures (organoids) and microsensor particles that enable assessment of cell vitality in real time by measuring oxygen concentration in a microfluidic system. The resulting TumOC platform allows for defined exposition of cytostatic drugs including dynamic and combination treatments. Measurements of cell vitality in real time enables analysis of not only the final effect of a drug treatment, but also the kinetics of drug response. At the same time, utilizing organoids allows for recapitulating tumor architecture and heterogeneity. Using the TumOC platform, we measured the cell vitality in real time over several days during treatment with classic chemotherapeutics or targeted cancer drugs and compared the results to end-point measurements on the same organoids in a static system. We assessed the effect of intra- and intertumoral heterogeneity and developed a protocol for treatment with clinically used combination regimes. In conclusion, our TumOC platform with its ability to recapitulate tumor heterogeneity in combination with dynamic treatments and real-time cell vitality assessment is an in-vitro tumor model closely recapitulating the physiological situation in a patient. TumOC provides an impressive opportunity to test and, consequently, predict the effectiveness of anti-cancer therapies. Therefore, this system is of interest not only for pre-clinical drug development but also for personalized oncology. Citation Format: Marie Flechner, Juergen Loskutov, Madeleine Nadolny, Ulrike Pfohl, Christian R. Regenbrecht, Katja Uhlig, Lena Wedeken. Personalized identification of cancer treatments in real-time: TumOC - a tumor organoid-on-chip platform for online cell vitality measurements [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 190.