Construction of Multi-Specific Antibody by Genetic Engineering and Its Progress in Tumor Therapy

Targeted treatment of cancer with monoclonal antibodies increases the benefit for patients. In order to improve the anti-tumor activity of monoclonal antibodies, multi-specific antibodies have entered the research field. The emergence of various techniques to produce multi-specific recombinant antibody molecules has led to the selection of target combinations in various forms. To date, only a few multi-specific constructs have entered phase III clinical trials, in contrast to classical monoclonal antibodies. Some of the format options are outlined from a technical point of view. We focus on the achievements and prospects of the underlying technologies for generating biand multispecific antibodies.

Multifaceted p21 in carcinogenesis,stemness of tumor and tumor therapy

Cancer cells possess metabolic properties that are different from those of benign cells.p21,encoded by CDKN1A gene,also named p21Cip1/WAF1,was first identified as a cyclin-dependent kinase regulator that suppresses cell cycle G1/S phase and retinoblastoma protein phosphorylation.CDKN1A(p21)acts as the downstream target gene of TP53(p53),and its expression is induced by wild-type p53 and it is not associated with mutant p53.p21 has been characterized as a vital regulator that involves multiple cell functions,including G1/S cell cycle progression,cell growth,DNA damage,and cell stemness.In 1994,p21 was found as a tumor suppressor in brain,lung and colon cancer by targeting p53 and was associated with tumorigenesis and metastasis.Notably,p21 plays a significant role in tumor development through p53-dependent and p53-independent pathways.In addition,expression of p21 is closely related to the resting state or terminal differentiation of cells.p21 is also associated with cancer stem cells and acts as a biomarker for such cells.In cancer therapy,given the importance of p21 in regulating the G1/S and G2 check points,it is not surprising that p21 is implicated in response to many cancer treatments and p21 promotes the effect of oncolytic virotherapy.

Clinical application of multidisciplinary teams in tumor therapy

Multidisciplinary team （MDT） model is a diagnostic and treatment model characterized by interdisciplinarity, integration, centralism, individualization, and precision and is becoming more common in the management of complex malignancies. MDT emphasizes team spirit and a personalized treatment strategy according to the actual condition of each patient. A cooperative and effective multidisciplinary team is an important guarantee for delivering high-quality services to patients. Under the guidance of a medical humanistic concept, MDT provides reasonable, effective, convenient, and a full range of excellent quality medical service to patients. The MDT maximizes patient benefits, and it is the developmental direction for large-scale general hospitals. At the same time, the MDT is also an important measure to strengthen the core competitiveness of hospitals. Here, we introduce the clinical application of the model in tumor therapy as well as the current state and development in our hospital.

Indocyanine green loaded graphene oxide for high-e±cient photoacoustic tumor therapy

Photoacoustic therapy,using the photoacoustic efect of agents for selectively kling tumor cells,has shown pronising for treat ing tumor.Utilization of high optical absorption probes can help to effectively improve the photoacoustic ther apy efficiency.Herein,we report a novel high-absorpt:ion photoacoustic probe that is composed of indocyanine green(ICG)and gr aphene oxide(GO),entitled GO-ICG,for photoacoustic ther apy.The attached ICG with narrow absorption spectral profile has strong optical absorption in the infrared region.The absorption spectrum of the GO-ICG solution reveals that the GO-ICG particles exhibited a 10-fold higher absorbance at 780 nm(its peak absorbance)as compared with GO.Importantly,ICG's fluorescence is quenched by GO via fuorescence resonance energy transfer.As a result,GO-ICG can high efficiently convert the absorbed light energy to acoustic wave under pulsed laser irradiation.We further demonstrate that GO-ICG can produce stronger photoacoustic wave than the GO and ICG alone.Moreover,we conjugate this contrast agent with integrin 0。As mono dlonal antibody to molecularly target the U87-MG human glioblastoma cells for selective tumor cell killing.Finally,our results testify that the photoacoustic therapy eficiency of GO-ICG is higher than the existing photoacoustic therapy agent.Our work demonstrates that GO-ICG is a high efficiency photo-acoustic therapy agent.This novel photoacoustic probe is likely to be an available candidate for tumor therapy.

Reductive damage induced autophagy inhibition for tumor therapy

Numerous therapeutic anti-tumor strategies have been developed in recent decades.However,their therapeutic efficacy is reduced by the intrinsic protective autophagy of tumors.Autophagy plays a key role in tumorigenesis and tumor treatment,in which the overproduction of reactive oxygen species(ROS)is recognized as the direct cause of protective autophagy.Only a few molecules have been employed as autophagy inhibitors in tumor therapy to reduce protective autophagy.Among them,hydroxychloroquine is the most commonly used autophagy inhibitor in clinics,but it is severely limited by its high therapeutic dose,significant toxicity,poor reversal efficacy,and nonspecific action.Herein,we demonstrate a reductive-damage strategy to enable tumor therapy by the inhibition of protective autophagy via the catalytic scavenging of ROS using porous nanorods of ceria(PN-CeO_(2))nanozymes as autophagy inhibitor.The antineoplastic effects of PN-CeO_(2)were mediated by its high reductive activity for intratumoral ROS degradation,thereby inhibiting protective autophagy and activating apoptosis by suppressing the activities of phosphatidylinositide 3-kinase/protein kinase B and p38 mitogen-activated protein kinase pathways in human cutaneous squamous cell carcinoma.Further investigation highlighted PN-CeO_(2)as a safe and efficient anti-tumor autophagy inhibitor.Overall,this study presents a reductive-damage strategy as a promising anti-tumor approach that catalytically inhibits autophagy and activates the intrinsic antioxidant pathways of tumor cells and also shows its potential for the therapy of other autophagy-related diseases.

Carrier-free nanoprodrug for p53-mutated tumor therapy via concurrent delivery of zinc-manganese dual ions and ROS

Human cancers typically express a high level of tumor-promoting mutant p53 protein(Mutp53)with a minimal level of tumor-suppressing wild-type p53 protein(WTp53).In this regard,inducing Mutp53 degradation while activating WTp53 is a viable strategy for precise anti-tumor therapy.Herein,a new carrier-free nanoprodrug(i.e.,Mn-ZnO_(2)nanoparticles)was developed for concurrent delivery of dual Zn-Mn ions and reactive oxygen species(ROS)within tumor to regulate the p53 protein for high anti-tumor efficacy.In response to the mild tumor acidic environment,the released Zn^(2+)and H_(2)O_(2)from Mn-ZnO_(2)NPs induced ubiquitination-mediated proteasomal degradation of Mutp53,while the liberative Mn^(2+)and increased ROS level activated the ATM-p53-Bax pathway to elevate WTp53 level.Both in vitro and in vivo results demonstrated that pH-responsive decomposition of Mn-ZnO2 NPs could effectively elevate the intracellular dual Zn-Mn ions and ROS level and subsequently generate the cytotoxic hydroxyl radical(·OH)through the Fenton-like reaction.With the integration of multiple functions(i.e.,carrier-free ion and ROS delivery,tumor accumulation,p53 protein modulation,toxic·OH generation,and pH-activated MRI contrast)in a single nanosystem,Mn-ZnO_(2)NPs demonstrate its superiority as a promising nanotherapeutics for p53-mutated tumor therapy.

Defect engineering endows NiTi stents with photothermal-enhanced catalytic activity for high-efficiency tumor therapy

NiTi stents are widely used in clinic for palliative care to relieve obstruction caused by Gastrointestinal(GI)cancers,which have high morbidity and mortality rates.However,tumor invasion and tumor overgrowth around the stent after surgery may lead to re-obstruction of the lumen.Thus,it is urgent to endow NiTi stents with excellent tumor suppressive ability and good biocompatibility.In this study,Ce-BTC was firstly prepared on the surface of NiTi pretreated by alkaline heat,followed by pyrolysis in Ar atmosphere at 450℃.Then,a composite coating consisting of defective cerium oxide and black Ni-Ti hydroxide/oxide was constructed on NiTi surface,which exhibited tumor microenvironment-response and hyperthermia-enhanced catalytic ability.Under near-infrared light irradiation,the photothermal performance of black Ni-Ti hydroxide/oxide and hyperthermia-enhanced catalytic activity of defective cerium oxide can achieve a synergistic effect of photothermal therapy and tumor catalytic therapy.Thereafter,defective cerium oxide can sustainably inhibit the proliferation of residual tumor cells by the generation of reactive oxygen species.Moreover,the composite coating has no obvious cytotoxicity to normal cells.This work provides a new insight for the preparation of stimulus-responsive antitumor stents for palliative treatment of gastrointestinal cancer.

Fabrication of microwave-sensitized nanospheres of covalent organic framework with apatinib for tumor therapy

A new nanocomposite of hollow covalent organic framework(COF)conjugated with the apatinib(AP)and loading microwave-sensitizer(ionic liquid,IL)was prepared by layer by layer(LBL)method and hyaluronic acid(HA)coating,named as COF-AP-IL@HA.AP loading rate in COF hollow-spheres(~30 nm shell thickness)was~40.3%,due to the interactions of hydrogen andπ-πbonds between AP and COF shell,and acidic environment destroyed COF structure,promoting AP release.Microwave sensitization of loaded IL in COF hollow-spheres could enhance the microwave heat-effect,and combined AP therapeutic ability,leading to their higher inhibitation on tumor,due to targeting ability of HA and the local release of apatinib.88.9%of inhibition rate of COF-AP-IL@HA under microwave on the in vivo tumor was significantly higher than those without microwave(12.3%)and COF-IL@HA with microwave(37.5%),indicating a synergism of sensitized microwave hyperthermia and AP therapy on the reduced expression of VEGF via the downregulation pathway of hypoxia inducible factor.These results indicated that COF-AP-IL@HA was potential to the application in the combination therapy of tumor of the sensitized microwave hyperthermia and apatinib.

Discovering metabolic vulnerability using spatially resolved metabolomics for antitumor small molecule-drug conjugates development as a precise cancer therapy strategy

Against tumor-dependent metabolic vulnerability is an attractive strategy for tumor-targeted therapy.However,metabolic inhibitors are limited by the drug resistance of cancerous cells due to their metabolic plasticity and heterogeneity.Herein,choline metabolism was discovered by spatially resolved metabolomics analysis as metabolic vulnerability which is highly active in different cancer types,and a choline-modified strategy for small molecule-drug conjugates(SMDCs)design was developed to fool tumor cells into indiscriminately taking in choline-modified chemotherapy drugs for targeted cancer therapy,instead of directly inhibiting choline metabolism.As a proof-of-concept,choline-modified SMDCs were designed,screened,and investigated for their druggability in vitro and in vivo.This strategy improved tumor targeting,preserved tumor inhibition and reduced toxicity of paclitaxel,through targeted drug delivery to tumor by highly expressed choline transporters,and site-specific release by carboxylesterase.This study expands the strategy of targeting metabolic vulnerability and provides new ideas of developing SMDCs for precise cancer therapy.

Nanoengineered sonosensitive platelets for synergistically augmented sonodynamic tumor therapy by glutamine deprivation and cascading thrombosis

Ultrasound(US)-activated sonodynamic therapy(SDT)stands for a distinct antitumor modality because of its attractive characteristics including intriguing noninvasiveness,desirable safety,and high tissue penetration depth,which,unfortunately,suffers from compromised therapeutic efficacy due to cancer cell-inherent adaptive mechanisms,such as glutathione(GSH)neutralization response to reactive oxygen species(ROS),and glutamine addictive properties of tumors.In this work,we developed a biological sonosensitive platelet(PLT)pharmacytes for favoring US/GSH-responsive combinational therapeutic of glutamine deprivation and augmented SDT.The amino acid transporter SLC6A14 blockade agentα-methyl-DL-tryptophan(α-MT)-loaded and MnO_(2)-coated porphyrinic metal-organic framework(MOF)nanoparticles were encapsulated in the PLTs through the physical adsorption of electrostatic attraction and the intrinsic endocytosis of PLTs.When the sonosensitive PLT pharmacytes reached tumor sites through their natural tendencies to TME,US stimulated the PLTs-loaded porphyrinic MOF to generate ROS,resulting in morphological changes of the PLTs and the release of nanoparticles.Subsequently,intracellular high concentration of GSH and extracellular spatio-temporal controlled US irradiation programmatically triggered the release ofα-MT,which enabled the synergistically amplified SDT by inducing amino acid starvation,inhibiting mTOR,and mediating ferroptosis.In addition,US stimulation achieved the targeted activation of PLTs at tumor vascular site,which evolved from circulating PLTs to dendritic PLTs,effectively blocking the blood supply of tumors through thrombus formation,and revealing the encouraging potential to facilitate tumor therapeutics.

“Block and attack”strategy for tumor therapy through ZnO_(2)/siRNA/NIR-mediating Zn^(2+)-overload and amplified oxidative stress

Intracellular zinc ion(Zn^(2+))accumulation disrupts the Zn^(2+)homeostasis,providing an ion-overloading anticancer strategy with great potential.The self-adaptation of tumor cells to ion concentration,however,puts forward higher requirements for the design of ion-overloading strategy.Herein,“block and attack”antitumor strategy was applied through a composite nanomaterials(UHSsPZH NPs).The strategy demonstrated powerful ion interference ability through both“blocking”the efflux of excess Zn^(2+)via gene silencing and“attacking”tumor cells via target delivery of ZnO_(2).After cellular internalization,ZnO_(2) was degraded to Zn^(2+)and hydrogen peroxide(H_(2)O_(2)),and the gene expression of zinc transporter 1(ZnT1)was silenced by targeting of released siRNA,which together caused intracellular Zn2+-overload.Disorder of Zn^(2+)further interfered with intracellular Ca^(2+)homeostasis,inhibited the electron transport chain and promoted the production of endogenous reactive oxygen species(ROS),which assisted the“attack”to tumor cells together with the exogenous ROS generated by UHSsPZH NPs under 980 nm laser irradiation.In summary,this work supplies a“block and attack”strategy for the application of ion homeostasis interference in tumor therapy.

Effect of endothelial progenitor cells in neovascularization and their application in tumor therapy

Objective To review the effect of endothelial progenitor cells in neovascularization as well as their application to the therapy of tumors.Data sources The data used in this review were mainly from PubMed for relevant English language articles published from 1997 to 2009. The search term was ＂endothelial progenitor cells＂.Study selection Articles regarding the role of endothelial progenitor cells in neovascularization and their application to the therapy of tumors were selected.Results Endothelial progenitor cells isolated from bone marrow, umbilical cord blood and peripheral blood can proliferate, mobilize and differentiate into mature endothelial cells. Experiments suggest endothelial progenitor cells take part in forming the tumor vascular through a variety of mechanisms related to vascular endothelial growth factor, matrix metalloproteinases, chemokine stromal cell-derived factor 1 and its receptor C-X-C receptor-4, erythropoietin, Notchsignal pathway and so on. Evidence demonstrates that the number and function change of endothelial progenitor cells in peripheral blood can be used as a biomarker of the response of cancer patients to anti-tumor therapy and predict the prognosis and recurrence. In addition, irradiation temporarily increased endothelial cells number and decreased the endothelial progenitor cell counts in animal models. Meanwhile, in preclinical experiments, therapeutic gene-modified endothelial progenitor cells have been approved to attenuate tumor growth and offer a novel strategy for cell therapy and gene therapy of cancer.Conclusions Endothelial progenitor cells play a particular role in neovascularization and have attractively potential prognostic and therapeutic applications to malignant tumors. However, a series of problems, such as the definitive biomarkers of endothelial progenitor cells, their interrelationship with radiotherapy and their application in cell therapy and gene therapy of tumors, need further investigation.

One-step nanoarchitectonics of a multiple functional hydrogel based on cellulose nanocrystals for effective tumor therapy

Hypoxia is a huge barrier for the development of photodynamic therapy(PDT).Chemodynamic therapy(CDT)could provide a possible solution to this dilemma.In this work,a controlled Schiff-base reaction was conducted between amido groups on the surface of carbon dots(CDs)and aldehyde groups on aldehyde-modified cellulose nanocrystals(mCNCs)as well as aldehydemCNCs decorated with Fe_(3)O_(4)nanoparticles.In this process,the mCNCs not only prevent the agglomeration of Fe_(3)O_(4)but also form hydrogels with CDs.The CDs act as both photothermal agent and photosensitizer.The hypoxia could be effectively relieved through the Fenton reaction due to the addition of Fe_(3)O_(4),and the·OH produced in the reaction further induces CDT and enhances tumor therapy efficiency.The therapy performance was further verified through in vitro cell experiments and in vivo animal experiments.This convenient method provides inspirations for the design and preparation of advanced biomaterials with multiple functions for cancer therapy.

Biomimetic copper single-atom nanozyme system for self-enhanced nanocatalytic tumor therapy

Single-atom nanozymes(SAZs)with peroxidase(POD)-like activity have good nanocatalytic tumor therapy(NCT)capabilities.However,insufficient hydrogen peroxide(H2O2)and hydrogen ions in the cells limit their therapeutic effects.Herein,to overcome these limitations,a biomimetic single-atom nanozyme system was developed for self-enhanced NCT.We used a previously described approach to produce platelet membrane vesicles.Using a high-temperature carbonization approach,copper SAZs with excellent POD-like activity were successfully synthesized.Finally,through physical extrusion,a proton pump inhibitor(PPI;pantoprazole sodium)and the SAZs were combined with platelet membrane vesicles to create PPS.Both in vivo and in vitro,PPS displayed good tumor-targeting and accumulation abilities.PPIs were able to simultaneously regulate the hydrogen ion,glutathione(GSH),and H2O2 content in tumor cells,significantly improve the catalytic ability of SAZs,and achieve self-enhanced NCT.Our in vivo studies showed that PPS had a tumor suppression rate of>90%.PPS also limited the synthesis of GSH in cells at the source;thus,glutamine metabolism therapy and NCT were integrated into an innovative method,which provides a novel strategy for multimodal tumor therapy.

Realize molecular surgical knife in tumor therapy by nanotechnology

In cancer therapy,the main challenge is how to attack the tumor and avoid injuring any normal organs in the meantime.In the last decades,scientists have made great efforts to try targeting the tumors,but little progress is achieved because all of the known therapeutic techniques could not latch the tumor cells down and always bring

Study of neutron dose equivalent at the HIRFL deep tumor therapy terminal

The secondary neutron fields at the deep tumor therapy terminal at HIRFL（Heavy Ion Research Facility in Lanzhou） were investigated. The distributions of neutron ambient dose equivalent were measured with a FHT762Wendi-II neutron ambient dose equivalent meter as ^（12）C ions with energies of 165, 207, 270, and 350 Me V/u were bombarded on thick tissue-like targets. The thickness of targets used in the experiments was larger than the range of the carbon ions. The neutron spectra and dose equivalent were simulated by using FLUKA code, and the results agree well with the experimental data. The experiment results showed that the neutron dose produced by fragmentation reactions in tissue can be neglected in carbon-ion therapy, even considering their enhanced biological effectiveness.These results are also valuable for radiation protection, especially in the shielding design of high energy heavy ion medical machines.

Melt electrospinning of daunorubicin hydrochloride-loaded poly (ε-caprolactone) fibrous membrane for tumor therapy

Daunorubicin hydrochloride is a cell-cycle non-specific antitumor drug with a high therapeutic effect.The present study outlines the fabrication of daunorubicin hydrochloride-loaded poly (ε-caprolactone)(PCL) fibrous membranes by melt electrospinning for potential application in localized tumor therapy.The diameters of the drug-loaded fibers prepared with varying concentrations of daunorubicin hydrochloride(1, 5, and 10 wt%) were 2.48 ± 1.25, 2.51 ± 0.78, and 2.49 ± 1.58 μm, respectively. Fluorescenceimages indicated that the hydrophobic drug was dispersed in the hydrophilic PCL fibers in theiraggregated state. The drug release profiles of the drug-loaded PCL melt electrospun fibrous membraneswere approximately linear, with slow release rates and long-term release periods, and no observed burstrelease. The MTT assay was used to examine the cytotoxic effect of the released daunorubicin hydrochlorideon HeLa and glioma cells (U87) in vitro. The inhibition ratios of HeLa and glioma cells followingtreatment with membranes prepared with 1, 5, and 10 wt% daunorubicin hydrochloride were 62.69%,76.12%, and 85.07% and 62.50%, 77.27%, and 84.66%, respectively. Therefore, PCL melt electrospun fibrousmembranes loaded with daunorubicin hydrochloride may be used in the local administration ofoncotherapy.

Phenylboronic acid-modified hollow silica nanoparticles for dual-responsive delivery of doxorubicin for targeted tumor therapy

This work reports a multifunctional nanocarrier based on hollow mesoporous silica nanoparticles(HMSNs)for targeting tumor therapy.Doxorubicin(DOX)was loaded into HMSNs and blocked with cytochrome C conjugated lactobionic acid(CytC–LA)via redox-cleavable disulfide bonds and pH-disassociation boronate ester bonds as intermediate linkers.The CytC–LA was used both as sealing agent and targeting motif.A series of characterizations demonstrated the successful construction of the drug delivery system.The system demonstrated pH and redox dual-responsive drug release behavior in vitro.The DOX loading HMSNs system displayed a good biocompatibility,which could be specifically endocytosed by HepG2 cells and led to high cytotoxicity against tumor cells by inducing cell apoptosis.In vivo data(tumor volume,tumor weight,terminal deoxynucleotidyl transferase dUTP nick end labeling and hematoxylin and eosin staining)proved that the system could deliver DOX to tumor site with high efficiency and inhibit tumor growth with minimal toxic side effect.

Study on the Anti-tumor Effect of Resveratrol Enhanced Photodynamic Therapy

Resveratrol,as a broad-spectrum anticancer agent,has significant inhibitory effects on many tumor cells such as murine liver cancer,human liver cancer,gastric cancer,breast cancer,ovarian cancer and leukemia.In this study,resveratrol was used as an inhibitor of ABCG2 to influence the efflux cells of protoporphyrin IX(PpIX)by inhibiting BCRP/ABCG2,and to investigate the effect of resveratrol on enhancing phototherapy(PDT)of human glioma cells treated with 5-aminolevulinic acid(5-ALA).Firstly,four kinds of malignant glioma cells U87MG,U251,A172 and T98G were treated with 5-ALA(1 mmol/L).Resveratrol at different concentrations(0.01~1.0μmol/L)as a modulator was incubated with glioma cells.The levels of PpIX,BCRP/ABCG2mRNA and ABCG2 protein in vivo and vitro and the effects of PDT in vitro were evaluated.The results showed that resveratrol decreased the expression of BCRP/ABCG2mRNA and thus decreased the expression of BCRP/ABCG2 protein on the cell membrane.Then,resveratrol at or above the concentration of 0.1μmol/L inhibited PpIX efflux in malignant brain glioma cells,and increased PpIX level in all cells with the increase of concentration.Resveratrol can not only inhibit the efflux of PpIX in malignant glioma cells,but also enhance the effect of PDT by increasing the content of PpIX in the cells.