Novel Multifunctional Nanocarriers For Targeted Drug/Gene Delivery

With the progress and development of nanomedicine,many nanocarriers have been widely used in the field of gene therapy or drug therapy for cancer.However,the existing gene and drug delivery vectors have defects such as complex preparation,high cost,and low targeting ability.In this paper,we constructed a dual-targeted nano-gene delivery platform based on polydopamine and a p H-based drug delivery platform based on nano graphene oxide（NGO）.The two specific contents are as follows:（1）Functionalized PDA/DEX-PEI@HA nanoparticles combining Sleeping-Beauty transposons deliver CRISPR/Cas9 system for gene therapy.In this work,CRISPR/Cas9 gene was cloned in a Sleeping-Beauty（SB）to obtain p T2Sp Cas9 for genome integration promotion.Meanwhile,PDA/DEX-PEI@HA（PDPH）nanoparticles,using polydopamine（PDA）as the carrier,hyaluronic acid（HA）as the cell-targeting ligand,and dexamethasone（DEX）as the nuclear localization signal（NLS）,were constructed for CRISPR/Cas9 targeting delivery.The results show that the particle size of PDPH/p T2Sp Cas9-Nanog was about 125 nm.At the cellular level,PDPH efficiently delivers p DNA in the cells and further into the nucleus.Remarkably,the cytotoxicity of PDPH was negligible in comparison to PEI_25K and PEI_10K.Western blots showed that after the transfection with PDPH/p T2Sp Cas9-Nanog/SB11,the Nanog protein in He La cells was completely knocked out,and the proliferation and migration abilities of tumor cells were decreased significantly.（2）Hydrazone-linked DOX-PEG-FA non-covalently modified nano graphene oxide for targeted drug delivery.In this work,we prepared DOX-hyd-PEG-FA polymers.Throughπ-πconjugation and hydrophobic interaction between DOX and NGO,the polymer was loaded on the surface of NGO to obtain NGO@DOX-hyd-PEG-FA nanodrugs.PEG in nanodrugs could significantly improve the biocompatibility of NGO.FA has improved the internalization rate of nanodrugs by targeting FA receptors on the surface of tumor cells.In the weakly acidic environment of the tumor,the hydrazone bond in nanodrug was decomposed and DOX was released into tumor tissues in large quantities.The results show that the particle size of NGO@DOX-hyd-PEG-FA was about 150 nm,and the drug Loading capacity of nanoparticles was more than 100%.which can effectively kill tumor cells while reducing the toxicity to normal cells.The drug release behavior,cell uptake ability,intracellular distribution,and in vitro cytotoxicity of NGO@DOX-hyd-PEG-FA were investigated.The results proved that NGO@DOX-hyd-PEG-FA nanoparticles have active targeting to tumor cells and can release large amounts of anticancer drugs in the weakly acidic environment of tumor,which can effectively kill tumor cells while reducing the toxicity to normal cells.