查找使用阿拉丁产品的科研文献

Fe3O4-Cy5.5-trastuzumab magnetic nanoparticles for magnetic resonance/near-infrared imaging targeting HER2 in breast cancer

Qiangqiang Yin,Xiaolong Gao,Hao Zhang,Zhichen Zhang,Xiaoyang Yu,Jialong He,Guangyue Shi,Liguo Hao

This study developed a probe Fe3O4-Cy5.5-trastuzumab with fluorescence and magnetic resonance imaging functions that can target breast cancer with high HER2 expression, aiming to provide a new theoretical method for the diagnosis of early breast cancer. Fe3O4-Cy5.5-trastuzumab nanoparticles were combined with Fe3O4 for T2 imaging and Cy5.5 for near-infrared imaging, and coupled with trastuzumab for HER2 targeting. We characterized the nanoparticles used transmission electron microscopy, hydration particle size, Zeta potential, UV and Fourier transform infrared spectroscopy, and examined its magnetism, fluorescence, and relaxation rate related properties. CCK-8 and blood biochemistry analysis evaluated the biosafety and stability of the nanoparticles, and validated the targeting ability of Fe3O4-Cy5.5 trastuzumab nanoparticles through in vitro and in vivo cell and animal experiments. Characterization results showed the successful synthesis of Fe3O4-Cy5.5-trastuzumab nanoparticles with a diameter of 93.72 ± 6.34 nm. The nanoparticles showed a T2 relaxation rate 42.29 mM−1s−1, magnetic saturation strength of 27.58 emg g−1. Laser confocal and flow cytometry uptake assay showed that the nanoparticles could effectively target HER2 expressed by breast cancer cells. As indicated by in vitro and in vivo studies, Fe3O4-Cy5.5-trastuzumab were specifically taken up and effectively aggregated to tumour regions with prominent NIRF/MR imaging properties. CCK-8, blood biochemical analysis and histological results suggested Fe3O4-Cy5.5-trastuzumab that exhibited low toxicity to major organs and good in vivo biocompatibility. The prepared Fe3O4-Cy5.5-trastuzumab exhibited excellent targeting, NIRF/MR imaging performance. It is expected to serve as a safe and effective diagnostic method that lays a theoretical basis for the effective diagnosis of early breast cancer. This study successfully prepared a kind of nanoparticles with near-infrared fluorescence imaging and T2 imaging properties, which is expected to serve as a new theory and strategy for early detection of breast cancer.

Optical biosensor based on weak value amplification for the high sensitivity detection of Pertuzumab in combination with Trastuzumab binding to the extracellular domain of HER2

Dongmei Li,Chaofan Weng,Chaoyi Chen,Kan Li,Qiang Lin,Yi Ruan,Yi Ruan,Jingjing Zhang,Shuqian Wang,Jia Yao,Jia Yao

A real-time optical phase sensing scheme based on weak value amplification was proposed to monitor the especially binding process of Pertuzumab combined with Trastuzumab on HER2 positive cells. From the wavelength shift of output spectrum, the phase difference between measuring and referential path related to the concentration of Pertuzumab as well as Trastuzumab could be calculated. With this approach, the limit of detection (LOD) of 5.54 × 10−13 M for Pertuzumab assay was achieved. Besides, the kinetics signal of Pertuzumab in combination with Trastuzumab binding to HER2 was detected in real time. Experimental results demonstrated that both Trastuzumab and Pertuzumab can be captured by HER2, but the former was significantly superior to the latter in terms of the target number. Additionally, the binding speed was analyzed and demonstrated to be closely correlated with the initial concentration of the targeting agents.

Mechanism exploration of Salvia miltiorrhiza Bunge against trastuzumab induced cardiotoxicity via multi-omics

Nuoxian Yu,Bo Cao,Jiahui Yang,Jing Xu,Mingyu Zhang,Xiaofei Fei,Zhuo Man,Jiabo Wang,Chunyu Li,Guohui Li

Ethnopharmacological relevance Despite the initial efficacy of trastuzumab (TRZ) in breast cancer treatment, its use is hampered by cardiotoxicity. Salvia miltiorrhiza Bunge (DS) has demonstrated potential in treating cardiovascular diseases, but its role in mitigating TRZ-induced cardiotoxicity remains unclear. Materials and methods We established both in vitro and in vivo models of TRZ-induced cardiotoxicity. Cell viability was measured using the CCK-8 assay following intervention with DS extract. The efficacy of varying doses of DS extract was assessed in the in vivo rat model. Cardiac function was determined via echocardiography, myocardial injury via biochemical markers, and tissue damage via haematoxylin and eosin (H&E) staining. Multi-omics integration (metabolomics, transcriptomics and network pharmacology) was used to identify potential therapeutic targets and bioactive compounds. We employed western blot analysis and immunofluorescence to confirm that DS extract modulates ferroptosis-related proteins. We used transmission electron microscopy (TEM) to observe the mitochondrial morphology in rat myocardial tissue. Then we employed molecular docking and dynamics simulations to validate the binding affinities of compounds to their targets. Results DS was found to exert cytotoxic effects on JIMT-1 breast cancer cells, as well as significantly ameliorating TRZ-induced myocardial injury in H9c2 rat cardiomyocytes in a dose-dependent manner. DS extract significantly improved cardiac function and reduced markers of myocardial injury in TRZ treated rats, while also attenuating histopathological damage. A combined analysis of transcriptomics, metabolomics and network pharmacology identified potential bioactive compounds and targets. Further validation revealed that DS extract treatment increased the expression of GPX4 and decreased the expression of ACSL4, thereby inhibiting lipid peroxidation and oxidative stress. TEM observations revealed that DS extract alleviates mitochondrial damage induced by TRZ. Tanshinone I, tanshinone II A, and tanshinone II B demonstrated a strong binding affinity for ACSL4 and GPX4 in molecular docking and dynamic simulations. Conclusion DS extract alleviates TRZ-induced cardiotoxicity, manifested by improved cardiac function, reduced myocardial injury markers, and mitigated tissue and mitochondrial damage. Its mechanism of action is associated with regulating proteins in the ferroptosis pathway (upregulating GPX4 and downregulating ACSL4). Additionally, tanshinone I, IIA, and IIB have been identified as potential active components.

Computational framework-guided engineering of microbial transglutaminase enables site-specific conjugation of Trastuzumab

Kanghui Yu,Zhe Wang,Xinxin Chen,Shuran Lei,Guodong Feng,Wei Zhang,Fangtian Fu,Anming Wang,Qiuyan Wang,Xiaolin Pei

A computational framework-guided strategy was developed to engineer MTGase rationally. • The mutant M3-MTGase-Sm exhibited a high catalytic activity and thermostability. • The mutant enabled site-specific conjugation at Q295 of the glycosylated antibody. • The approach offers a promising enzymatic route to specifically modify antibodies. Microbial transglutaminase (MTGase) specifically catalyzes the conjugation between primary amine-containing molecules and glutamine residues, making it a promising enzymatic approach for producing homogeneous antibody-drug conjugates (ADCs). However, this strategy faces challenges associated with glycan-induced steric hindrance, insufficient enzyme activity, and poor thermal stability. In this study, we engineered MTGase from Streptomyces mobaraensis (MTGase-Sm) using a computational framework-guided strategy to enhance its catalytic cc for site-specific conjugation at the Q295 site of trastuzumab. A triple mutant M3-MTGase-Sm (R48K/G283S/S303G) exhibited a catalytic activity of 21.1 U/mg toward the model substrate CBZ-Gln-Gly, representing a 3.6-fold improvement over the wild-type enzyme. Furthermore, the variant demonstrated enhanced thermostability. Its half-life at 50 °C was 65 min, approximately a 1.4-fold increase over that of the wild-type MTGase-Sm. Structural and molecular dynamics simulations further revealed the underlying mechanism. These mutations regulate the local structural flexibility of the enzyme, stabilize the geometric configuration of the active center, and optimize long-range conformational dynamics, thereby effectively improving enzymatic properties. The engineered M3-MTGase-Sm can conjugate small-molecule probes (dansylcadaverine) and polyethylene glycol derivatives (mPEG-NH 2 ) to the conserved glutamine residue Q295 of trastuzumab. Notably, it maintains improved conjugation performance even in the presence of native N-glycans at the adjacent sterically hindered N297 site. Nevertheless, this variant has not yet been validated for conjugation with cytotoxic agents. This study establishes a computation-informed framework for MTGase evolution, and the engineered MTGase demonstrates significant potential for the biosynthesis of ADCs with high specificity and uniformity. Download: Download high-res image (152KB) Download: Download full-size image

PolyA-mediated interfacial assembly of probes for label-free electrochemical detection of trastuzumab

Zheng Zhao,Yuheng Liao,Haidong Li,Jia-Huan Qu,JingHao Wang,Qiqin Wang,Zhengjin Jiang

PolyA-mediated spatial regulation enables modification-free probe assembly on gold electrodes. • An optimized AAP30 spacer promotes an ordered interfacial probe arrangement for efficient antibody recognition. • The engineered interface integrates label-free electrochemical sensing with reduced cellular adhesion. • The platform supports sensitive trastuzumab detection in serum and advances therapeutic antibody analysis. The analytical performance of heterogeneous biosensors is critically governed by the interfacial arrangement of recognition probes, as non-directional immobilization often limits binding-site exposure and hinders the recognition of bulky biomacromolecules. Herein, an interfacial engineering strategy based on the nanoscale spatial regulation of modification-free probes was developed to construct a label-free electrochemical biosensor for the detection of therapeutic antibodies. Using a polyA-aptamer probe (AAP) composed of the antibody-specific aptamer and the terminal polyA segment, we achieved the oriented self-assembly of probes onto gold electrodes through sequence-dependent nucleobase‑gold affinity, eliminating the need for complex chemical coupling. By adjusting the length of the polyA segment, the interfacial organization of the probes was systematically regulated. Results demonstrated that an optimized AAP30 probe facilitated a transition from an overcrowded, disordered monolayer to a well-spaced, near-upright architecture. This rational interfacial arrangement provides sufficient steric freedom for probe reconfiguration, thereby enhancing target accessibility and epitope recognition. Furthermore, the highly hydrated, polyanionic DNA layer contributed to improved surface wettability and reduced cellular adhesion, supporting the applicability of the biosensor in biological matrices. Using trastuzumab as a model therapeutic antibody, the proposed platform exhibited sensitive, label-free detection with a wide linear range from 10 ng/mL to 100 μg/mL and a limit of detection (LOD) of 5.05 ng/mL. Overall, this polyA-mediated spatial regulation strategy offers a practical and adaptable platform for designing label-free electrochemical biosensors for therapeutic drug monitoring in complex biological samples. Download: Download high-res image (104KB) Download: Download full-size image

Trastuzumab-functionalized bionic pyrotinib liposomes for targeted therapy of HER2-positive breast cancer

Du Jiaqun,Liu Xiaobang,Sun Junpeng,Wu Qian,Hu Yu,Shi Huan,Zheng Li,Liu Ying,Wu Chao,Gao Yu

In this study, we prepared a bionic nanosystem of trastuzumab-functionalized SK-BR-3 cell membrane hybrid liposome-coated pyrotinib (Ptb-M-Lip-Her) for the treatment of HER2-positive breast cancer. Transmission electron microscopy, dynamic light scattering, polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting were used to verify the successful preparation of Ptb-M-Lip-Her. In vitro drug release experiments proved that Ptb-M-Lip-Her had a sustained release effect. Cell uptake experiments and in vivo imaging experiments proved that Ptb-M-Lip-Her had good targeting ability to homologous tumor cells (SK-BR-3). The results of cell experiments such as MTT, flow cytometry, immunofluorescence staining and in vivo antitumor experiments showed that Ptb-M-Lip-Her could significantly promote apoptosis and inhibit the proliferation of SK-BR-3 cells. These results clearly indicated that Ptb-M-Lip-Her may be a promising biomimetic nanosystem for targeted therapy of HER2-positive breast cancer.

Trastuzumab modified silica nanoparticles loaded with doxorubicin for targeted and synergic therapy of breast cancer

Ling-xin Meng,Qiang Ren,Qin Meng,Yu-xiu Zheng,Mao-lei He,Shu-yan Sun,Zhao-jun Ding,Bing-cheng Li,Hui-yun Wang

Untargeted delivery as well as low efficacy are two main obstacles for effective breast cancer therapy. Here in this study, we surface modified silica nanoparticles (SLN) with Trastuzumab (Tra) to construct a tumor-targeting carrier (Tra-SLN) for specific drug delivery to human epidermal growth factor receptor 2 (HER2) overexpressing breast cancer cells. In addition, Tra-SLN could also loaded with broad-spectrum anticancer drug doxorubicin (DOX) to finally construct a drug delivery system (DDS) capable of co-delivering Tra and DOX (Tra-SLN/DOX). Our results demonstrated that the as-prepared Tra-SLN/DOX was nanoscale particles with spheroid appearance which showed preferable stability in physiological environments. In addition, the Tra-SLN/DOX could specifically target to HER2 overexpressed MCF-7 cells. Both in vitro and in vivo experiments revealed that the Tra-SLN/DOX exerted enhanced anticancer efficacy when compared with Tra or DOX alone. It was suggested that Tra-SLN/DOX might be a promising platform for enhanced therapy of breast cancer.

Targeted inhibition of HER-2 positive breast cancer cells by trastuzumab functionalized pullulan-doxorubicin nanoparticles

Ruiling Xu,Junhui Sui,Mingda Zhao,Yuedi Yang,Lei Tong,Yongmei Liu,Yong Sun,Yujiang Fan,Jie Liang,Xingdong Zhang

Human epidermal growth factor receptor-2 (HER-2) positive breast cancer is highly invasive with poor clinical outcomes and high risk of recurrence. Here, trastuzumab functionalized pullulan-doxorubicin nanoparticles (Tz-P-Dox) were developed and characterized as a new anticancer formulation for active targeting HER-2 overexpression of breast cancer cells. The obtained nanoparticles had the hydrodynamic diameter of 66.7 ± 2.0 nm and PDI of 0.218 ± 0.012. And the in vitro results showed significant difference of cell uptake and cytotoxic effect between Tz-P-Dox and non-targeted P-Dox against HER-2 positive cell lines (BT474 and MCF-7 cells). However, the differences between Tz-P-Dox and P-Dox were not observed in HER-2 negative cell lines (MDA-MB-231 cells). These results suggested that trastuzumab functionalized nanoparticles had great potential to be considered as a candidate drug for HER-2 positive breast cancer treatment.

Trastuzumab potentiates doxorubicin-induced cardiotoxicity via activating the NLRP3 inflammasome in vivo and in vitro

Shanshan Wei,Wanjun Ma,Yuanying Yang,Taoli Sun,Chuanhao Jiang,Jian Liu,Bikui Zhang,Wenqun Li

Trastuzumab (Tra), the first humanized monoclonal antibody that targets human epidermal growth factor receptor 2 (HER2), is commonly used alongside doxorubicin (Dox) as a combination therapy in HER2-positive breast cancer. Unfortunately, this leads to a more severe cardiotoxicity than Dox alone. NLRP3 inflammasome is known to be involved in Dox-induced cardiotoxicity and multiple cardiovascular diseases. However, whether the NLRP3 inflammasome contributes to the synergistic cardiotoxicity of Tra has not been elucidated. In this study, primary neonatal rat cardiomyocyte (PNRC), H9c2 cells and mice were treated with Dox (15 mg/kg in mice or 1 μM in cardiomyocyte) or Tra (15.75 mg/kg in mice or 1 μM in cardiomyocyte), or Dox combined Tra as cardiotoxicity models to investigate this question. Our results demonstrated that Tra significantly potentiated Dox-induced cardiomyocyte apoptosis and cardiac dysfunction. These were accompanied by the increased expressions of NLRP3 inflammasome components (NLRP3, ASC and cleaved caspase-1), the secretion of IL-β and the pronounced production of ROS . Inhibiting the activation of NLRP3 inflammasome by NLRP3 silencing significantly reduced cell apoptosis and ROS production in Dox combined Tra-treated PNRC. Compared with the wild type mice, the systolic dysfunction , myocardial hypertrophy , cardiomyocyte apoptosis and oxidative stress induced by Dox combined Tra were alleviated in NLRP3 gene knockout mice . Our data revealed that the co-activation of NLRP3 inflammasome by Tra promoted the inflammation, oxidative stress and cardiomyocytes apoptosis in Dox combined Tra-induced cardiotoxicity model both in vivo and in vitro . Our results suggest that NLRP3 inhibition is a promising cardioprotective strategy in Dox/Tra combination therapy.

Targeted delivery of polyamidoamine-paclitaxel conjugate functionalized with anti-human epidermal growth factor receptor 2 trastuzumab

Pengkai Ma,Xuemei Zhang,Ling Ni,Jinming Li,Fengpu Zhang,Zheng Wang,Shengnan Lian,Kaoxiang Sun

Comparative study of trastuzumab modification analysis using mono/multi-epitope affinity technology with LC-QTOF-MS

Chengyi Zuo,Jingwei Zhou,Sumin Bian,Qing Zhang,Yutian Lei,Yuan Shen,Zhiwei Chen,Peijun Ye,Leying Shi,Mao Mu,Jia-Huan Qu,Zhengjin Jiang,Qiqin Wang

Dynamic tracking analysis of monoclonal antibodies (mAbs) biotransformation in vivo is crucial, as certain modifications could inactivate the protein and reduce drug efficacy. However, a particular challenge (i.e. immune recognition deficiencies) in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen. To address this limitation, a multi-epitope affinity technology utilizing the MOF@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region (CDR) mimotope peptide (HH24) and non-CDR mimotope aptamer (CH1S-6T) onto the surface of MOF@Au nanocomposite. Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology. Moreover, the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and LC-QTOF-MS. Therefore, multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb. Particularly, the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids. Compared to the spiked PB model, faster modification trends were monitored in the spiked serum and patients’ sera due to the catalytic effect of plasma proteins and relevant proteases. Differences in peptide modification levels of trastuzumab in patients’ sera were also monitored. In summary, the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb, contributing to improved understanding and paving the way for future research and clinical applications.

Osthole attenuates trastuzumab-induced cardiotoxicity in mice by enhancing autophagy via regulating the p38MAPK/mTOR signaling pathway

Huan Hou,Yaping Yang,Minyan Zhu,Meilin Xie,Yan Zhang,Rong Chen

Background and aim Trastuzumab (TRZ)-induced cardiotoxicity impairs breast cancer patients' quality of life, with no effective prevention currently. Osthole (OST) has been reported to exert cardioprotective effects in certain cardiovascular diseases. The purpose of this study was to investigate the probable mechanisms behind the protective effect of OST against TRZ-induced cardiotoxicity. Experimental procedure Mice were pretreated with OST (50 or 100 mg/kg) for 5 days, followed by TRZ (10 mg/kg, 10 days) to establish a cardiotoxicity model. The protective effects of OST on TRZ-induced cardiotoxicity was assessed via echocardiography, cardiac weight index (CWI), serum myocardial injury markers, H&E, and Masson staining. To explore the mechanisms, SOD/MDA levels, DHE staining (ROS), Nick-End-Labeling (TUNEL) staining (apoptosis), and Western blot were performed with/without 3-methyladenine (autophagy inhibitor) or SB203580 (p38MAPK inhibitor). Results and Conclusion OST pretreatment significantly improved cardiac function, alleviated myocardial damage and fibrosis, and enhanced autophagy (increased LC3II/I, Beclin-1 and decreased p62). Oxidative stress damage was ameliorated by OST, as demonstrated by higher activity of SOD, lower level of MDA and reactive oxygen species (ROS). In addition, OST decreased the ratio of Bax/Bcl-2 and Caspase-3 expression, and reduced apoptosis rate. OST activated p38MAPK (increased p-p38MAPK level) while inhibiting mTOR (decreased p-mTOR level). While these effects were blocked by the autophagy inhibitor 3-MA or p38MAPK inhibitor SB203580. These results indicated that OST has a protective effect on TRZ-induced cardiotoxicity, and its underlying mechanism may be related to enhancing autophagy via the p38MAPK/mTOR signaling pathway, thus reducing oxidative stress and apoptosis.