Unraveling new strategies for early detection of invasive cancer cells: A special focus on the nuclear and cytoskeletal architecture

Abstract Introduction: Cancer invasion is a hallmark of cancer progression and the leading cause of cancer mortality worldwide. However, the identification of invasive cancer cells at an early stage remains challenging. Therefore, there is an urgent need to develop novel biomarkers and innovative strategies to successfully tackle cancer invasion. It is well established that the structure and organization of the nucleus and the cytoskeleton are dynamically orchestrated during many cellular processes, including cancer invasion. Thus, in this study, we investigated fine-tuned nuclear and cytoskeletal patterns associated with invasion, taking into consideration the cell-extracellular matrix (ECM) interaction. Methods: In this study, we took advantage of an ECM Array platform (MicroMatrix 36, MicroStem) and a panel of invasive and non-invasive gastric cancer cells. For validation purposes, we assessed cells transfected with wild type or mutant E-cadherin, which lead to distinct adhesion and invasion phenotypes. A comprehensive analysis of nuclei and cytoskeletal architectural features was performed. Nuclei area, intensity, perimeter, eccentricity, circularity, solidity and entropy were evaluated. Notably, a new computational pipeline was developed to characterize cytoskeletal structures, namely orientation, compactness, radiality, bundling, parallelness, morphology, among others. Results and Discussion: Our results have shown that the invasive abilities of cancer cells are associated with unique nuclear and cytoskeletal modifications. Moreover, we verified that invasive cells adhere preferentially to specific ECM components. Interestingly, we could observe an association between nuclear phenotypes and increased attachment abilities, suggesting that the ECM may modulate nuclear morphometrics thus affecting the invasive behavior of cancer cells. Of note, cell lines harboring E-cadherin mutations corroborated the existence of a nuclear signature associated with invasive potential. Remarkably, the proposed computational framework has shown that the microtubules of cells with disrupted E-cadherin are shorter, have uniform length patterns and are more compactly distributed as compared with cells with wild-type E-cadherin. Conclusion: Overall, this study demonstrates that the use of nuclear and cytoskeletal architectural features could provide an efficient strategy to identify cells with invasive potential. Further investigation and integration of this approach with other cellular properties could have an impact in diagnosis, prognosis and therapeutic strategies. Citation Format: Maria Sofia Fernandes, Joana Figueiredo, Diogo Vieira, Soraia Melo, Ana Margarida Moreira, João Miguel Sanches, Raquel Seruca. Unraveling new strategies for early detection of invasive cancer cells: A special focus on the nuclear and cytoskeletal architecture [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 3348.