APPLIED AND ENVIRONMENTAL MICROBIOLOGY
·
2019
·Vol. 85
·期 12
·
IF 4.077
Xinyu Lu,Weiwei Wang,Lige Zhang,Haiyang Hu,Ping Xu,Tao Wei,Hongzhi Tang
N,N-Dimethylformamide (DMF) is one of the most common xenobiotic chemicals, and it can be easily emitted into the environment, where it causes harm to human beings. Herein, an efficient DMF-degrading strain, DM1, was isolated and identified as Methylobacterium sp. This strain can use DMF as the sole source of carbon and nitrogen. Whole-genome sequencing of strain DM1 revealed that it has a 5.66-Mbp chromosome and a 200-kbp megaplasmid. The plasmid pLVM1 specifically harbors the genes essential for the initial steps of DMF degradation, and the chromosome carries the genes facilitating subsequent methylotrophic metabolism. Through analysis of the transcriptome sequencing data, the complete mineralization pathway and redundant gene clusters of DMF degradation were elucidated. The dimethylformamidase (DMFase) gene was heterologously expressed, and DMFase was purified and characterized. Plasmid pLVM1 is catabolically crucial for DMF utilization, as evidenced by the phenotype identification of the plasmid-free strain. This study systematically elucidates the molecular mechanisms of DMF degradation by Methylobacterium.IMPORTANCE DMF is a hazardous pollutant that has been used in the chemical industry, pharmaceutical manufacturing, and agriculture. Biodegradation as a method for removing DMF has received increasing attention. Here, we identified an efficient DMF degrader, Methylobacterium sp. strain DM1, and characterized the complete DMF mineralization pathway and enzymatic properties of DMFase in this strain. This study provides insights into the molecular mechanisms and evolutionary advantage of DMF degradation facilitated by plasmid pLVM1 and redundant genes in strain DM1, suggesting the emergence of new ecotypes of Methylobacterium.
研究领域:
环境微生物学
·
生物化学
✓ 已验证
DOI
高影响因子
Drug Design Development and Therapy
·
2022
·
IF 4.319
Keying Liang,Shengsheng Mei,Xiangzheng Gao,Shanshan Peng,Jinbiao Zhan
Purpose T-DM1 is an antibody–drug conjugate (ADC) consisting of trastuzumab and DM1 linked together. T-DM1 binds to human epidermal growth factor receptor-2 (HER2) in tumors and then triggers the endocytosis of T-DM1 and release of payload. Therefore, endocytosis efficacy is considered as a critical step for the initiation of T-DM1 therapy; however, the endocytosis mechanism of T-DM1 remains poorly understood. Meanwhile, HER2 is regarded as an internalization-resistant receptor, which hinders the endocytosis and effectiveness of T-DM1. The present study is to explore the T-DM1 endocytosis pathway, which may provide insights into the internalization mechanism of ADCs and help to improve efficacy.Methods Confocal microscopy and flow cytometry were used to analyse T-DM1 intracellular trafficking and endocytosis efficiency, while Western blot assay was performed to detect T-DM1 degradation.Results We found that intracellular T-DM1 was increased to 50% within 12 h. T-DM1 was colocalized with cholera toxin B (CTxB), a lipid raft marker, within 2 h and then degraded in lysosome. Upon overexpression of caveolin-1 (CAV-1) and utilization of caveolae/lipid-raft disruptors, we found that temporal CAV-1 upregulation significantly facilitated T-DM1 endocytosis and degradation, whereas nystatin and lovastatin disrupted caveolae/lipid-raft structure and inhibited T-DM1 degradation. We demonstrate that T-DM1 internalizes through the lipid raft-mediated endocytosis in a CAV-1 dependent manner, rather than through the clathrin-mediated endocytosis in HER2-positive cancer cells.Conclusion Our findings suggest that modulation of the caveolae/lipid-raft mediated endocytosis may be a possible option for improving the clinical therapeutic effect of T-DM1 because it plays a key role in regulating T-DM1 internalization.
研究领域:
肿瘤学
·
细胞生物学
·
药理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
RSC Advances
·
2023
·Vol. 13
·期 16
·10840-10846
·
IF 3.9
Qiu-Yan Zhang,Qing-Long Yu,Wei-Jing Luan,Tong-Fang Li,Ya-Ni Xiao,Li Zhang,Yi Li,Rong Rong,Chun-Guang Ren
Colorectal cancer (CRC) is one of the most prevalent cancers worldwide as well as a significant cause of mortality. The conventional treatment could cause serious side effects and induce drug resistance, recurrence and metastasis of cancers. Hence, specific targeting of cancer cells without affecting the normal tissues is currently an urgent necessity in cancer therapy. The emerging of peptide–drug conjugates (PDC) is regarded as a promising approach to address malignant tumors. LWJ-M30, a conjugate of DM1 and B6 peptide, targeted transferrin receptors (TfRs) on the surface of the CRC cells, showing a powerful anti-cancer effect. LWJ-M30 significantly inhibited the HCT116 cells proliferation and migration in vitro. LWJ-M30 also dramatically decreased the level of polymeric tubulin, while the disruption of microtubules caused the cell cycle to be arrested in the G2/M phase. LWJ-M30 induced the HCT116 cells apoptosis both in vivo and in vitro. The results in vivo demonstrated that LWJ-M30 could inhibit the HCT116 growth without affecting the mouse body weight. Taking these results together, our data indicated that LWJ-M30 could improve the therapeutic effects of DM1 while reducing the systemic toxicity in normal tissues.
研究领域:
肿瘤学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2016
·
IF 4.712
Hua Sang,Gaoyuan Lu,Yazhong Liu,Qiyue Hu,Weifeng Xing,Dongbing Cui,Fang Zhou,Jingwei Zhang,Haiping Hao,Guangji Wang,Hui Ye
Trastuzumab-MCC-DM1 (T-DM1) is an antibody-drug conjugate (ADC) that consists of a monoclonal antibody (mAb) trastuzumab non-cleavably linked to a cytotoxic drug DM1. During production, the DM1 agents were conjugated to the lysine residues of the mAb in a non-specific manner, yielding a heterogeneous mixture of ADC molecules that differ with respect to both the number and the conjugation sites of DM1 per mAb molecule. Since drug conjugation sites of ADC can significantly impact properties such as stability and pharmacokinetic behaviors, a rapid and reliable approach for conjugation site analysis of ADCs is highly demanded. Herein, we have employed a signature ion fingerprinting approach to specifically determine lysine residues with DM1 conjugation, and developed a normalized peak area quantitation method to characterize the percentage of DM1-conjugated lysine for each putative site using a T-DM1 biosimilar as a model drug. With this integrative approach, 38 lysine residues were identified with DM1 conjugation among 90 possible sites. More interestingly, we found that the T-DM1 biosimilar exhibited a specific preference of DM1-conjugation for several lysine residues, and such preference was consistent among three production batches. A molecular modeling approach was subsequently utilized to analyze all the conjugation sites, and revealed an intriguing correlation of the conjugated residue's microenvironment with the conjugation level. In summary, our study introduced an approach that is widely applicable to ADCs of interest for conjugation site analysis. Moreover, it suggests the necessity of performing conjugation site analysis for product and process characterization and also for routine use in lot release and stability testing of manufactured ADCs.
研究领域:
生物化学
·
药物分析
·
蛋白质组学
✓ 已验证
DOI
高影响因子
Journal of Translational Medicine
·
2022
·Vol. 20
·期 1
·1-10
·
IF 8.44
Huang Kun,Wang Dan-Dan,Hu Wen-Bao,Zeng Wei-Qian,Xu Xia,Li Qiu-Xiang,Bi Fang-Fang,Yang Huan,Qiu Jian
Background Myotonic dystrophy type 1 (DM1), one of the most common forms of adult-onset muscular dystrophy, is caused by abnormally expanded CTG repeats in the 3′ untranslated region of the DMPK gene. The CUG repeats transcribed from the expanded CTG repeats sequestrate a splicing factor, MBNL1, causing the clinical symptoms in DM1. Nowadays, only symptomatic treatments are available for DM1, and no rational therapy is available. Recently, upregulation of MBNL1 expression has been found to be one of the promising therapies for DM1.Methods All experiments were conducted in the C2C12 myoblasts and HSALR mice, a DM1 mouse model. Real-time PCR and western blot were used to detect the mRNA and protein level, respectively. The rotarod exercise, grip strength and hanging time were used to evaluate the muscle strength of mice.Results In this study, we demonstrated that calcitriol, an active form of vitamin D3, increased MBNL1 in C2C12 mouse myoblasts as well as in HSALR mice model for DM1. In HSALR mice model, calcitriol improved muscle strength, and corrected aberrant splicing in skeletal muscle. Besides, calcitriol reduced the number of central nuclei, and improved muscle histopathology in HSALR mice. In addition, we identified that calcitriol upregulated MBNL1 expression via activating the promoter of Mbnl1 in C2C12 myogenic cells.Conclusion Our study suggests that calcitriol is a potential pharmacological strategy for DM1 that enhances MBNL1 expression.
研究领域:
分子生物学
·
神经病学
·
药理学
·
肌肉骨骼疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PROTEOME RESEARCH
·
2024
·
IF 3.8
Meiling Qi,Chenyue Zhu,Yi Chen,Chenxi Wang,Xinyuan Ye,Sen Li,Zhongzhe Cheng,Hongliang Jiang,Zhifeng Du
Antibody–drug conjugate (ADC) consists of engineered antibodies and cytotoxic drugs linked via a chemical linker, and the stability of ADC plays a crucial role in ensuring its safety and efficacy. The stability of ADC is closely related to the conjugation site; however, no method has been developed to assess the stability of different conjugation sites due to the low response of conjugated peptides. In this study, an integrated strategy was developed and validated to assess the stability of different conjugation sites on ADC in serum. Initial identification of the conjugated peptides of the model drug ado-trastuzumab emtansine (T-DM1) was achieved by the proteomic method. Subsequently, a semiquantitative method for conjugated peptides was established in liquid chromatography–hybrid linear ion trap triple quadrupole mass spectrometry (LC–QTRAP-MS/MS) based on the qualitative information. The pretreatment method of the serum sample was optimized to reduce matrix interference. The method was then validated and applied to evaluate the stability of the conjugation sites on T-DM1. The results highlighted differences in stability among the different conjugation sites on T-DM1. This is the first study to assess the stability of different conjugation sites on the ADC in serum, which will be helpful for the design and screening of ADCs in the early stages of development.
研究领域:
药学
·
分析化学
·
生物化学
·
药物开发
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Qiyue Wang,Xiaofei Gu,Ji Liu,Yan Zhao,Zhen Li,Shuyan He,Dong Shen,Xiaofei Xin,Yan Shen
Glioblastoma (GBM) is the most common malignant primary brain tumor, with an overall median survival of less than two years after resection. The urgent goal in pre-clinical investigations is to develop effective therapeutic strategies to prolong the GBM patients’ survival and improve the poor prognosis. Although various inorganic nanoplatforms have been applied in GBM treatment, they still face obstacles such as poor brain targeting, low tumor effective penetration, and serious monotherapy resistance. Here, we designed an environmental-dependent tumor-killing nanoreactor (RFcAD) with a gold-ferri nanoagglomerates form structure that effectively inhibits GBM proliferation via catalyzing reactive oxygen species (ROS) generation, exhausting glucose, and NIR-based photothermal therapy. With iRGD decoration, RFcAD can effectively penetrate the blood–brain barrier and target GBM. The “bomb-like” structured RFcAD exhibited a glutathione (GSH)-responsive release of toxicity agent DM1 and pH-dependent nanoagglomerates degradation with gold particles and superparamagnetic iron oxide nanoparticles (SPIONS) release. Gold particles and SPIONS destroy GBM cells as chemodynamic therapy agents via exhibiting a glucose oxidase (GOx)-like and peroxide (POx)-like catalytic ability to generate cytotoxic ROS and exhaust intracellular glucose, respectively. Synergistic with DM1-based chemotherapy and NIR-induced photothermal therapy, RFcAD can significantly reduce GBM proliferation both in vitro and in vivo. This work highlights that the RFcAD nanoreactor can benefit GBM inhibition by improving tumor targeting, tissue penetration, and cytotoxicity.
研究领域:
纳米技术
·
癌症治疗
·
生物医学工程
·
药学
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）