Time, the final frontier

Abstract

Cancer's notorious heterogeneity poses significant challenges, as each tumor comprises a unique ecosystem. While single-cell and spatial transcriptomics advancements have transformed our understanding of spatial diversity within tumors, the temporal dimension remains underexplored. Tumors are dynamic entities that continuously evolve and adapt, and relying solely on static snapshots obscures the intricate interplay between cancer cells and their microenvironment. Here, we advocate for integrating temporal dynamics into cancer research, emphasizing a fundamental shift from traditional endpoint experiments to data-driven, continuous approaches. This integration involves, for instance, the development of advanced live imaging techniques, innovative temporal omics methodologies, and novel computational tools.

Keywords: cancer heterogeneity; circadian clock; live‐cell imaging; metastasis; temporal dynamics; temporal omics.

Fig. 1

Time‐dependent measurements identify crucial biological phenomena. This figure highlights the importance of temporal resolution in capturing dynamic biological processes across various assays. (A) Dynamics of endothelial cell junctions: Live imaging using a Zeiss LSM880 Airyscan microscope (40×, water immersion) reveals that endothelial cell junctions—critical for blood vessel integrity—open and close rapidly. A single imaging plane is shown (scale bar: 3 μm). (B) Tensin‐1 condensates in cancer cells: Using spinning disk microscopy (63×, oil immersion), this panel tracks intracellular tensin‐1 (GFP‐tagged) in cancer cells over 10 min with images every 10 s. The left panel compares the first and last time points, while the right panel employs a temporal color code (Fire LUT) across all frames to highlight dynamic changes. A maximum intensity projection is provided (scale bar: 3 μm). (C) Endothelial monolayer permeability assays: Impedance measurements (monitor barrier function) were performed on a confluent endothelial monolayer after treatment. The left graph shows the start and end results (triplicates), while the right graph presents continuous measurements every 15 min for 24 h. Compounds 1 and 2 exhibit opposite effects and distinct kinetics, demonstrating the limitations of the endpoint assays. Error bars represent the standard deviation. (D) Circulating tumor cells attachment to endothelial cells: Live widefield imaging (20×; NA 0.45, Nikon CFI S Plan Fluor ELWD) of cancer cells perfused over endothelial monolayers in a microfluidic system. Cancer cells were individually tracked. The curves of the three selected cells illustrate different behaviors: green cells stably adhere at first contact, while orange and blue cells exhibit metastable adhesions before complete arrest. High temporal resolution is essential for studying these mechanisms. Scale bar: 50 μm. (E) Migrating cancer cells: Live imaging of a cancer cell expressing Paxillin‐mEmerald (PXN‐mEm) migrating on a micropatterned extracellular matrix track (100×; NA 1.4 oil, Zeiss Plan‐Apochromat). The top row shows selected time points of the cell's central position, while the bottom row depicts intermediate migration stages. An 8‐h recording with 20‐min intervals captures the cell's active exploration of its environment. Scale bars: 25 μm. (A–E) Data presented in this figure are for illustrative purposes only and are derived from one biological repeat.