Research Projects

The rationale for this study stems from the critical need to enhance the efficacy of CD8+ T cells in cancer immunotherapy, where early activation and sustained functionality are essential for effective tumor eradication. Current approaches often fail to maintain CD8+ T cell potency due to premature senescence driven by hyperactivation of the mTOR pathway following initial expansion. By targeting eukaryotic elongation factor-2 kinase (eEF2K), a key regulator of cellular stress responses, in combination with mTOR inhibition, this research aims to overcome these limitations, ensuring prolonged T cell activity and increased cytotoxicity. This dual-target strategy could revolutionize the way we enhance immune responses, providing a novel therapeutic avenue to improve cancer immunotherapy outcomes. Loss of eEF2K in CD8+ T cells promote early proliferation but leads to accelerated senescence due to the sustained hyperactivation of the mTOR signaling pathway. This study hypothesizes that targeting eEF2K can optimize early proliferation and improve the cytotoxic function of CD8+ T cells. Additionally, we hypothesize that combining eEF2K inhibition with mTOR inhibition will further enhance CD8+ T cell function and persistence. To test this hypothesis, we will identify the most effective eEF2K inhibitors through in-silico docking and screening, followed by in vitro testing to determine optimal inhibitors and dosages. Preliminary findings have identified three potential candidates: A484954, Oleuropin, and Thieno-pyridine 2,3 carboxylic acid. Notably, Oleuropin also inhibits mTOR, making it a promising dual inhibitor for both pathways. In our primary experiment, we will isolate and activate CD8+ T cells from OT-1 mice, treating them with the optimal eEF2K inhibitor and mTOR inhibitor (rapamycin) to assess their proliferation, cytokine production, and functional responses. Mechanistic studies will explore signaling pathways activated by this dual inhibition, including NF-κB and MAP-kinase. Finally, we will evaluate the therapeutic potential of these treated CD8+ T cells in vivo by injecting them into B16-ovalbumin melanoma-bearing mice and assessing their effects on tumor growth and survival. This research has the potential to significantly advance cancer immunotherapy by optimizing CD8+ T cell functionality through targeted inhibition of key signaling pathways. By simultaneously inhibiting eEF2K and mTOR, we aim to overcome the limitations of early proliferation and senescence in CD8+ T cells, enhancing their cytotoxic activity and antitumor efficacy. The results from this study could not only provide novel therapeutic strategies to boost T cell responses in cancer but also offer insights into the broader mechanisms of immune cell longevity and function. Ultimately, this work could lead to more effective and durable immune-based treatments for cancer, paving the way for improved patient outcomes and survival.