Engineered Cell Membrane-Derived Nanovesicles For Precise Tumor Therapy

Over the past decades,in response to the heterogeneity of malignant tumors and the individualized treatment needs of patients,precise tumor therapy has received more and more attention.The tumor precision treatment efficiency can be impoved by combination of nano drug delivery system and precise tumor therapy at the molecular level in terms of targeted drug delivery,theranostics,and controlled drug release.Cell membrane-derived nanovesicles have been widely used in precise tumor therapy due to their personalized biological origin.At present,the application research of cell membrane-derived nanovesicles in the precise tumor therapy mainly focuses on the improvement of drug targeted delivery capabilities.However,due to the limitations of the existing cell membrane-derived nanovesicles functionalization strategies,the low yield,low drug loading,poor controllability of loading method and uncontrollable drug release hinder the development of theranostics based on cell membrane-derived nanovesicles.In this paper,electroporation and membrane fusion strategies are used to develop theranostics and controlled drug release systems based on cell membrane-derived nanovesicles,and achieved high-efficient tumor therapy.The main research results of this paper are as follows:(1)A multifunctional tumor cell membrane derived microparticles drug delivery system is developed by electroporation functionalization strategy for imaging guided synergistic photothermal/low-dose chemotherapy of tumor.In this study,bismuth selenide nanodots and doxorubicin hydrochloride co-embedded multifunctional tumor cell-derived microparticles(Bi₂Se₃/DOX@MPs)are fabricated by ultraviolet light irradiation-induced budding of parent cells which were preloaded with theranostics reagents bismuth selenide nanodots and doxorubicin hydrochloride via electroporation.This electroporation construction strategy is efficient and controllable,and Bi₂Se₃/DOX@MPs are obatained high drug-loading capacity and high yield without unfavorable damage to the membrane structure,maintaining their excellent inherent biological behavior.Intracellular uptake studies have indicated that the cellular internalization of Bi₂Se₃/DOX@MPs is mainly mediated through membrane fusion,which is not only beneficial to enhancing cell uptake and tumor penetration capabilities of Bi₂Se₃/DOX@MPs,but also achiving high efficiency killing of tumor cells in vitro without considering endosomal escape.The multifunctional microparticles also exhibit high tumor targeting and photothermal heating effect in vivo due to their inherent tumor homing targeting ability.The real-time non-invasive visual monitoring of the accumulation of drugs in the tumor tissue realized through the excellent CT and PA imaging performance of Bi₂Se₃/DOX@MPs,and can be used to guide the subsequent treatment.Under the guidance of CT/PA dual-modality imaging,the synergistic anti-tumor effect of photothermal therapy and low-dose chemotherapy is successfully realized,and an excellent tumor treatment effect is achieved.In addition,the low hemolytic activity,metabolizability and low systemic toxicity of Bi₂Se₃/DOX@MPs imply their good biocompatibility.(2)A hybrid erythrocyte membrane-derived nanovesicles drug delivery system is constructed by membrane fusion functionalization strategy for sonodynamic trigging drug release and enhanced tumor targeting therapy.In this study,erythrocyte membrane-derived nanovesicles and cationic liposomes composed of unsaturated phospholipids and loaded with hematoporphyrin monomethyl ether and doxorubicin is prepared by ultrasonic extrusion method and film dispersion method,respectively.Through membrane fusion,ultrasound responsive drug-loaded hybrid membrane vesicles(DOX/HMME@FA-NL)are constructed.The reliability and effectiveness of the membrane fusion strategy are confirmed through characterization of particle size and zeta potential,F?rster energy resonance transfer and fluorescence co-localization analysis.Under ultrasound stimulation,high levels of reactive oxygen species are produced in vitro.The unsaturated phospholipids in the hybrid membrane vesicles can be oxidized via reactive oxygen species,leading to the destroying of the structure of hybrid membrane vesicles to achieve the controlled release of drugs,thereby enhancing their tumor cell killing effect.The linkage of the folate targeting group also enhances the tumor targeting ability of DOX/HMME@FA-NL.After H22 tumor-bearing mice are intravenous injected with DOX/HMME@FA-NL and treated with ultrasound,they achieve better than expected tumor sonodynamic responsive treatment effects.Based on the construction strategies of electroporation and membrane fusion,the engineered cell membrane-derived nanovesicles are fabricated via controllable methods.Compared with traditional construction methods,the drug-loaded amount and yield of nanovesicles are increased,and the controllable release of drugs in tumor tissues is realized,which enhancing the precise tumor therapy efficiency.