High-order Aggregation Of Lipid Rafts By Self Assembled DNA Origami Inhibits Cancer Metastasis

Cell migration plays a central role in various biological phenomena from embryogenesis to tumor metastasis.Currently,the main method for studying the regulation of cell migration behavior is ligand induced aggregation of receptors on the surface of cell membranes.However,there is a single receptor involved in the study of regulating cell migration,and the receptor aggregation process relies on random collisions and nondirected migration of receptors on the cell membrane,resulting in low efficiency in the regulation of receptor aggregation.Therefore,in the study of cell migration,it is vital to develop a strategy to regulate multi-receptor aggregation and improve the efficiency of receptor aggregation.Some studies have shown that the proteins exist in the lipid rafts on the surface of the cell membrane are closely related to the formation of the cytoskeleton and the migration behavior of cells.However,the small size and dynamic characteristics of lipid rafts provides challenges for precise regulation of lipid rafts.Due to its flexible structural design,DNA origami nanotechnology has great potential for accurately assembling molecules at the nanoscale.In this study,we developed a DNA origami self-assembly technique on the surface of cell membranes.It can efficiently anchor the lipid rafts on the cell membrane and mediate two-dimensional planar high-order aggregation of the lipid rafts.This technology can significantly inhibit the migration of cancer cells in vitro and in vivo,and is applied to cancer treatment.In addition,we have explored the interrelationship between lipid rafts and cell migration behavior,laying a foundation for studying the cellular function of lipid rafts in cell biology.The DNA origami based self-assembly technology proposed in this work has great potential in regulating the functional domains of cell membranes and guiding cell function or behavior.The main content of this paper is as follows:Chapter 1 OverviewFirstly,this chapter introduces the research progress of DNA molecular assembly technology from three dimensions of DNA molecule assembly size: one-dimensional DNA molecular assembly,two-dimensional DNA molecular assembly,and threedimensional DNA molecular assembly.Secondly,this chapter introduces the biological application of DNA molecular assembly technology in the functionalization of three membrane surfaces: living cell membranes,extracellular vesicles,and synthetic lipid vesicles.Finally,the research significance and content of this work are summarized.Chapter 2 Construction of DNA origami self-assembly technology on cell membrane surfaceIn this chapter,we developed a DNA origami based self-assembly technique for specifically and efficiently targeting lipid rafts on cell membranes to induce highorder aggregation of lipid rafts.We have designed the staple strands of DNA origami to provide anchoring and self-assembly capabilities.In order to simulate the environment and behavior of phospholipid bilayer on cell membranes,a new method for supporting lipid bilayer DNA origami self-assembly based on the surface of mica sheets was established,enabling DNA origami to exhibit a two-dimensional interface assembly morphology different from that in a solution environment.In addition,the self-assembly of DNA origami on cell membranes still has significant biocompatibility.This work provides a powerful tool and platform for the controllable self-assembly of functional domains on the surface of cell membranes.Chapter 3 Self-assembly of DNA origami on cell membrane surface to inhibit cell migration studyIn this chapter,we investigated the expression and ability of DNA origami selfassembly technology developed to inhibit the migration of cancer cells in vitro and in vivo.In vitro,we studied the migration behavior of clustered cells and the motility of individual cells.The results showed that DNA origami induced lipid raft aggregation can significantly inhibit the migration of cancer cells in vitro.The study of DNA origami self-assembly in vivo using a mouse tumor model also demonstrated its excellent ability to inhibit tumor growth and migration.Proteomic analysis has been used to demonstrate the molecular mechanism by which DNA origami self-assembly inhibits cell migration behavior.This work demonstrates the significant potential of regulating lipid rafts on the surface of cell membranes by utilizing the structural design advantages of DNA origami molecular tools in inhibiting the migration behavior of cancer cells.