Spatiotemporal Control Over Multicellular Migration Using Green Light Reversible Cell–Cell Interactions

Abstract

cell-cell adhesions leads to the progression of cancer [1c] and subsequent metastasis. [4] Additionally, the control of cell-cell interactions is of interest in the field of bottomup tissue-engineering, which aims to assemble cells as the basic unit into functional tissues. [5] Precise control in time and space over cell-cell interactions is required to successfully assemble appropriate multicellular architectures that resemble the in vivo structures and to control how cells work together within a tissue.In recent years, many tools have been developed to control the formation and the disassembly of cell-cell interactions in a controlled manner and thereby have given insight into the role of cell-cell adhesions. [6] For instance, chemical modifications of the plasma membrane with bioorthogonal reactive functional groups [7] and specific noncovalent interaction partners [8] including complementary DNA strands, [9] biotin-streptavidin, [10] and supramolecular binding partners [11] result in the formation of chemical bonds between neighboring cells. However, in only a few examples, it is possible to reverse these cell-cell interactions once formed. [11] To overcome general concerns related to the chemical modification of the cell membrane (e.g., degradation over time, off-target cell toxicity), it is possible to regulate the expression and the activity of genetically encoded native cell-cell adhesion molecules such as cadherins [12] or artificial surface receptors. [1d,9b,6,12,13] These adhesions allow cells to not only just be brought together but in some cases also to transduce intracellular signals. [1e,12,14] Photoregulation of cell-cell adhesions provides high spatiotemporal control, since light, as a stimulus, can be focused on the desired area and delivered at any given time. Using photocleavable nitrobenzyl [15] and switchable azobenzyl [14] chemical linkers, it has been shown possible to control cell-cell interactions with UV light. [11b,15,16] More recently, optogenetic tools for the regulation of cell-cell interactions with visible light have improved the biocompatibility, [13b,e] while also making it possible to dynamically and reversibly control cell-cell interactions between multiple cell types. [13b,e,16b,17] In these reports, photoswitchable proteins were expressed on the cell's plasma membrane as artificial adhesion molecules, which induced cell-cell interactions through the light-dependent dimerization of these proteins. [6] Depending on the photoswitchable proteins employed cell-cell adhesions between the same or differentThe regulation of cell-cell adhesions in space and time plays a crucial role in cell biology, especially in the coordination of multicellular behavior. Therefore, tools that allow for the modulation of cell-cell interactions with high precision are of great interest to a better understanding of their roles and building tissuelike structures. Herein, the green light-responsive protein CarH is expressed at the plasma membrane of cells as an artificial cell adhesion rece...

Opto-E-cadherin: Optogenetic control over cell-cell adhesions

“…For migration assays in the dark, a long pass filter (514/LP edge basic long pass filter, BLP01-514R-25 from AHF Analysentechnik) was placed in front of the white light source. The wound closure rate was determined with an established script in ImageJ, 25 and the migration angle and correlation length were obtained by using MATLAB version 7.10 (R2020a) with the PIVlab -particle image velocimetry (PIV) application developed by Dr. William Thielicke and Prof. Eize J. Stamhuis (MATLAB script: supplementary data). 48 For spatiotemporal control in the wound-healing assay, the cells were prepared as described earlier, and the experiment was conducted on a confocal microscope (Leica SP8) at 37 °C under 5% CO2.…”

“…[18][19][20] The current state of the art involves an optochemical, 21 and optogenetic tools that allows light-induced dissociation of adherens junctions in through photocleavable linkers, 22 photoswitchable small molecules introduced through metabolic labelling, 23 and artificial photoswitchable surface proteins that mediate artificial cellcell interactions. [24][25][26][27] Yet, these tools are limited by the need for light-responsive chemicals, lack of reversibility, use of UV light or the fact that they do not link to E-cadherin-associated cell signalling.…”

Engineering Light-Control in Biology

“…Nzigou Mombo et al exploited that surface-displayed CarH requires AdoB12 to form a tetramer, and consequently used the AdoB12 concentration in the media (0.5–10 μm) as a tuning knob for the ratio of active CarH. Cells started clustering at concentrations of 1 µm AdoB12 and the area of the clusters increased with the concentration ( Nzigou Mombo et al, 2021 ). Apart from exogenous supplementation, it was shown that de novo engineering of vitamin B12 synthesis in E. coli via an aerobic biosynthetic pathway is possible ( Fang et al, 2018 ).…”

An Adenosylcobalamin Specific Whole‐Cell Biosensor

“…[28] CarH is an extremely versatile light-responsive protein, which besides its applications in optogenetics to control receptor activity [29] and gene expression, [30,31] has also been employed to generate light-responsive hydrogels [32] and protein micro-patterning. [33,34] Moreover, when it is expressed as a surface receptor, it is able to mediate light-responsive cell-cell adhesions in mammalian cells, [35] which makes it a suitable candidate to also mediate bacterial cell-cell adhesions.…”