Collective Cell Migration in 3D Epithelial Wound Healing

Abstract

Collective cell migration plays a pivotal role in development, wound healing, and metastasis, but little is known about the mechanisms and coordination of cell migration in 3D microenvironments. Here, we demonstrate a 3D wound healing assay by photothermal ablation for investigating collective cell migration in epithelial tissue structures. The nanoparticle-mediated photothermal technique creates local hyperthermia for selective cell ablation and induces collective cell migration of 3D tissue structures. By incorporating dynamic single cell gene expression analysis, live cell actin staining, and particle image velocimetry, we show that the wound healing response consists of 3D vortex motion moving toward the wound followed by the formation of multicellular actin bundles and leader cells with active actin-based protrusions. Inhibition of ROCK signaling disrupts the multicellular actin bundle and enhances the formation of leader cells at the leading edge. Furthermore, single cell gene expression analysis, pharmacological perturbation, and RNA interference reveal that Notch1-Dll4 signaling negatively regulates the formation of multicellular actin bundles and leader cells. Taken together, our study demonstrates a platform for investigating 3D collective cell migration and underscores the essential roles of ROCK and Notch1-Dll4 signaling in regulating 3D epithelial wound healing.

Nrf2 modulates the hybrid epithelial/mesenchymal phenotype and Notch signaling during collective cancer migration

“…Notch signaling has been shown to control collective cell migration. 19,22,23,[30][31][32][33] We therefore measured the distributions of Notch markers (i.e., Notch1, Dll4, and Jagged1) in the WT and under Nrf2 perturbations (Figure 4A-C). In agreement with previous studies, 19,23 Notch signaling was upregulated near the leading edge.…”

Development of an Enhanced-Throughput Radial Cell Migration Device

“…Cellular motility is an imperative aspect of tissue generation, organ formation, wound healing, and inflammatory responses, among many other physiological functions that are essential for life. [21][22][23][24] While many fields of biological research have significant interest in cellular migration, it is cemented as a hallmark of cancer and considered one of the key cellular functions that enable metastatic spread, which ultimately causes mortality. In essentially all tumor types, metastasis is the single most important identifier of patient mortality.…”

Development of a high-throughput radial migration device

“…Migration and motility are potentially the most crucial phenotypic characteristics cells intrinsically harbor. Cellular motility is an imperative aspect of tissue generation, organ formation, wound healing, inflammatory responses, among many other physiological functions that are essential for human survival (17)(18)(19)(20). While many fields of biological research have significant interest in cellular migration, it is cemented as a hallmark of cancer and pinned as one of the key cellular functions which enable metastatic spread and ultimately patient mortality.…”