Nano-vesicles Targeting Receptor Overexpressing Tumors For Drug Delivery On The Precise Diagnosis And Therapy Of Hepatobiliary Tumors

Inspired by natural biology,bioengineering-modified cell membrane camouflaging nanotechnology has been widely investigated.Many researchers have devoted themselves to developing site-specific therapeutic agent-delivery systems.At present,various tumor active-targeting ligands(protein,peptide,full-length monoclonal antibodies)have been used for genetically modified membrane vesicles,such as human sodium taurocholate co-transporting polypeptide,affibody,site-specific targeting peptide for human sodium taurocholate co-transporting polypeptide,aspargineglycine-arginine peptide,and human monoclonal antibodies antiglypican 3.This basic research provided guidance for the prevention,diagnosis and treatment of liver diseases including hepatitis B virus infection and hepatobiliary tumors.Additionally,cell membrane-derived vesicles have attracted much attention as a carrier for small molecules owing to their superiorities of high bio-compatibility and encapsulation efficiency,controllable size,easy genetic modification and great flexibility to encapsulate various bio-active molecules.Wrapping near-infrared fluorescent dye in bio-engineered nano-vesicles can significantly improve the effective drug concentration for legible imaging in guiding cancer treatment.Thus,this nano-vesicles-based nano-biotechnology offers a broad prospect in combining natural functionalities with active targeting ligands and provides a means of enhancing effective drug concentration in the tumor for personalized treatment in the future nano-medicine.In the further studies,we explored the development and application of bioengineering-modified cell membrane nano-vesicles.This study includes two parts.The result of first part is that bio-engineered cell membrane nanovesicles with Her2-specific affibody mediated tumor targeting delivery of indocyanine green(ICG)that enhanced the multimodal imaging-guided photothermal therapy against Her2-positive perihilar cholangiocarcinoma(pHCC).Herein,we engineered a human epidermal growth factor receptor 2(Her2)affibody to the surface of cell membrane nanovesicles(A-Nvs)in a ligand-oriented manner and loaded them with ICG(A-Nvs@ICG)as precision theranostics for pHCC treatment.In vitro,the NIR fluorescence signal(2.67±0.15)times,photoacoustic signal(1.25± 0.23)times and photothermal signal(1.15± 0.32)times of A-Nvs@ICG were higher than those of ICG,respectively.In vivo,the NIR fluorescence imaging signal and photoacoustic imaging signal in the A-Nvs@ICG group were(5.18 ± 3.2)times and(4.16±1.32)times higher than those in the ICG group,respectively,which is used to guide the complete ablation of tumors by photothermal therapy.The A-Nvs@ICG were prepared and exhibited satisfactory targeting effects and good biosafety in Her2-overexpressing pHCC cells.In vivo fluorescence and photoacoustic imaging demonstrated that A-Nvs@ICG promoted the accumulation of ICG in pHCC tissue,leading to enhanced tumor regression and improved anti-cancer effects when combined with photoirradiation.Therefore,bio-engineered A-Nvs@ICG represent a promising nanotheranostic agent for pHCC with potential for clinical translation.The result of second part is that bio-engineered cell membrane nanovesicles with growth arrest-specific 6(Gas6)protein mediated tumor targeting delivery of IR820 that enhanced the multimodal imaging-guided surgical navigation and photothermal therapy against Axl-positive hepatic carcinoma.New indocyanine green(IR820)showed excellent properties in imaging and photothermal conversion.Long-term unsolved health problems from pre-/intra-/postoperative complications and thermal ablation complications pose threats to liver-cancer patients.Using a bioengineering approach,G-Nvs@IR820,a kind of human-cell-membrane nano-vesicle,was generated with Gas6 expressed on the membrane and with IR820 loaded into it(G-Nvs@IR820),which is proven to be an effective nanoparticle-drug-delivery system for Axl-overexpressing hepatocellular carcinoma.In vitro,the NIR fluorescence signal(6.76±1.44)times,photoacoustic signal(1.25 ± 0.14)times,surgical navigation fluorescence signal(8.09 ± 2.55)times,and photothermal signal(1.17±0.68)times were higher than those of IR820,respectively.In vivo,compared with IR820,the specific binding of Gas6 to Axl enhanced the IR820 delivery to tumor of G-Nvs@IR820,resulting in the increase of the high-resolution of NIR fluorescence imaging signal and photoacoustic imaging signal in tumors by(2.96±2.11)times and(2.68 ± 0.61)times,respectively.On one hand,G-Nvs@IR820 has good targeting ability to the tumor site and also has a good ability to guide the further accurate obliteration of carcinoma from adjacent normal tissue in surgery with its highly resolved surgical-navigation signals.On the other hand,the G-Nvs@IR820 represented a new perspective for photothermal therapy.As Gas6 binds to Axl specifically,G-Nvs@IR820 has good targeting ability to the tumor site and also has a good ability to guide the further accurate obliteration of carcinoma from adjacent normal tissue in surgery with its highly resolved fluorescence/photoacoustic/surgical-navigation signals.In addition,G-Nvs@IR820 show high biosafety and low toxicity.In brief,bio-engineered nano-vesicles for drug delivery,show high affinity,specificity,and safety.And it has a good prospect for clinical application.