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.

ANALYSIS FOR A FREE BOUNDARY PROBLEM MODELING TUMOR THERAPY

This paper is devoted to studying a free boundary problem modeling the effects of drug resistance and vasculature on the response of solid tumors to therapy.The model consists of a system of partial differential equations governing intra-tumoral drug concentration and cancer cell density.By applying the Lp theory of parabolic equations and the Banach fixed point theorem,it is proved that this problem has a unique global classical solution.

Three positive charge nonapoptotic-induced photosensitizer with excellent water solubility for tumor therapy

Photodynamic therapy(PDT)has emerged as a significant cancer therapy option.Currently,cation-based organic small molecule aggregation-induced emission(AIE)photosensitizers(PSs)attract the wide atten-tion of many scientists,due to improved reactive oxygen species(ROS)production after cationization.However,such PSs tend to localize only the mitochondria,limiting the death way of tumor cells(usu-ally apoptosis)during PDT process,which may affect the therapeutic effect under some circumstances.Herein,we designed a novel water-soluble three positive charge PS,TPAN-18F,which could be distributed uniformly in cell cytoplasm and had distribution in different sub-organelles(mitochondria,endoplasmic reticulum,lysosome).The experimental results showed that TPAN-18F-based PDT process can not only disrupt mitochondrial functions(reducing ATP production and destroying mitochondrial membrane po-tential),but also elevate the intracellular lipid peroxides(LPOs)level,which evoke the non-apoptotic death manner of tumor cells.Further,in vivo studies showed that TPAN-18F-based PDT could effectively inhibit tumor growth.Accordingly,we believe that the construction of TPAN-18F is suggestive for tumor non-apoptotic therapy.

CA IX-targeted Ag_(2)S quantum dots bioprobe for NIR-II imaging-guided hypoxia tumor chemo-photothermal therapy

Hypoxia is the common characteristic of almost all solid tumors,which prevents therapeutic drugs from reaching the tumors.Therefore,the development of new targeted agents for the accurate diagnosis of hypoxia tumors is widely concerned.As carbonic anhydrase IX(CA IX)is abundantly distributed on the hypoxia tumor cells,it is considered as a potential tumor biomarker.4-(2-Aminoethyl)benzenesulfonamide(ABS)as a CA IX inhibitor has inherent inhibitory activity and good targeting effect.In this study,Ag_(2)S quantum dots(QDs)were used as the carrier to prepare a novel diagnostic and therapeutic bioprobe(Ag_(2)S@polyethylene glycol(PEG)-ABS)through ligand exchange and amide condensation reaction.Ag_(2)S@PEG-ABS can selectively target tumors by surface-modified ABS and achieve accurate tumor imaging by the near infrared-II(NIR-II)fluorescence characteristics of Ag_(2)S QDs.PEG modification of Ag_(2)S QDs greatly improves its water solubility and stability,and therefore achieves high photothermal stability and high photothermal conversion efficiency(PCE)of 45.17%.Under laser irradiation,Ag_(2)S@PEG-ABS has powerful photothermal and inherent antitumor combinations on colon cancer cells(CT-26)in vitro.It also has been proved that Ag_(2)S@PEG-ABS can realize the effective treatment of hypoxia tumors in vivo and show good biocompatibility.Therefore,it is a new efficient integrated platform for the diagnosis and treatment of hypoxia tumors.

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.

Ferritin-based drug delivery system for tumor therapy

Ferritin has been widely used in drug delivery in recent years due to its unique spherical nanocage structure.In addition,the natural targeting and good biocompatibility of ferritin make it an excellent drug delivery system.Therefore,ferritin has shown a broad application prospect in the field of nanomedicine.In this perspective,we will describe ferritin-based drug delivery system,ferritin-based drug delivery system for tumor treatment,and future challenges in ferritin development.Hopefully,this perspective may inspire the future development of ferritin-based drug delivery systems for tumor treatment.

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.

Intelligent micro/nanorobots for improved tumor therapy

A major bottleneck underlying nanomaterial-based tumor therapy lies in complex biological environment and physiological barriers.Micro/nanorobots with the features of self-propulsion and controllable navigation have gradually become a research hotspot in the tumor therapeutic community,exhibiting their advantages in efficient cargo loading,controllable cargo delivery,stimulitriggered cargo release,deeper tumor tissue penetration,and enhanced cargo accumulation in tumor tissue.In this review,the self-propulsion and controllable navigation are introduced as two major properties of micro/nanorobots,in which micro/nanorobots are propelled by chemical reactions,physical fields,and biological systems and could be navigated by chemotaxis,remote magnetic guidance,and light.Then,the recent advances of micro/nanorobots for chemotherapy,immunotherapy,photothermal therapy,photodynamic therapy,chemodynamic therapy,and multimodal tumor therapy would be discussed.Finally,the perspective and challenges are also mentioned.It is expected that this review gives an insight into intelligent micro/nanorobots for improved tumor therapy,aiming for more extensive and in-depth investigations,and final applications in the clinic.

“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.

Influencing factors and solution strategies of chimeric antigen receptor T-cell therapy(CAR–T)cell immunotherapy

Chimeric antigen receptor T-cesll therapy(CAR–T)has achieved groundbreaking advancements in clinical application,ushering in a new era for innovative cancer treatment.However,the challenges associated with implementing this novel targeted cell therapy are increasingly significant.Particularly in the clinical management of solid tumors,obstacles such as the immunosuppressive effects of the tumor microenvironment,limited local tumor infiltration capability of CAR–T cells,heterogeneity of tumor targeting antigens,uncertainties surrounding CAR–T quality,control,and clinical adverse reactions have contributed to increased drug resistance and decreased compliance in tumor therapy.These factors have significantly impeded the widespread adoption and utilization of this therapeutic approach.In this paper,we comprehensively analyze recent preclinical and clinical reports on CAR–T therapy while summarizing crucial factors influencing its efficacy.Furthermore,we aim to identify existing solution strategies and explore their current research status.Through this review article,our objective is to broaden perspectives for further exploration into CAR–T therapy strategies and their clinical applications.

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.

Real-time in vivo visualization of tumor therapy by a near-infrared-II Ag2S quantum dot-based theranostic nanoplatform

Real-time and objective feedback of therapeutic efficacies would be of great value for tumor treatment. Here, we report a smart Ag2S QD-based theranostic nanoplatform （DOX@PEG-Ag2S） obtained by loading the anti-cancer drug doxorubicin （DOX） into polyethylene glycol-coated silver sulfide quantum dots （PEG-Ag2S QDs） through hydrophobic-hydrophobic interactions, which exhibited high drug loading capability （93 wt.% of DOX to Ag2S QDs）, long circulation in blood （t1/2 = 10.3 h）, and high passive tumor-targeting efficiency （8.9% ID/gram） in living mice where % ID/gram reflects the probe concentration in terms of the percentage of the injected dose （ID） per gram of tissue. After targeting the tumor tissue, DOX from PEG-AgRS cargoes was selectively and rapidly released into cancer cells, giving rise to a significant tumor inhibition. Owing to the deep tissue penetration and high spatio-temporal resolution of Ag2S QDs fluorescence in the second near-infrared window （NIR-II）, the DOX@PEG-Ag2S enabled real-time in vivo reading of the drug targeting process and therapeutic efficacy. We expect that such a novel theranostic nanoplatform, DOX@PEG-Ag2S, with integrated drug delivery, therapy and assessment functionalities, will be highly useful for personalized treatments of tumors.