Frontiers in Cellular and Infection Microbiology
·
2025
·
IF 4.8
Jiansheng Lin,Chunyan Lin,Jingyang Zheng
Background: P. diazotrophicus was isolated from a newborn with D-galactosemia complicated sepsis. A homologous blaCAR-2 gene, encoding CAR-2, a predicted member of the CAR family subclass B3 metallo-β-lactamases (MBLs), was found in the genome of this strain. This study aimed to explore the identification of a novel CAR-2 protein encoded by the chromosome of P. diazotrophicus Pd1 that exhibits the zinc-binding motifs of subclass B3 enzymes and the regulatory pattern of CARR, located directly upstream of the blaCAR-2 gene, on the blaCAR-2 gene and its impact on antibiotic resistance.Methods: Antibiotic susceptibility testing was conducted by the plate agar dilution method. Site-directed mutagenesis was conducted using Mut Express II Fast Mutagenesis Kit V2. Kinetic assays were used to determine the hydrolysis of β-Lactam. The construction of bacterial knockout strains was carried out according to the principle of homologous recombination. The detection of the mRNA expression level of the gene was performed by Real-time Quantitative PCR (qPCR).Results: The minimum inhibitory concentrations (MICs) for E.coli DH5α(pHSG398::CAR-2), which expressed blaCAR-2, increased significantly for cefalothin, cefuroxime and cefotaxime sodium by 2-, 16- and 32- fold, respectively, which showed that blaCAR-2 had resistance to these three antibiotics. This protein, which was a MBL, contains two classical zinc-binding sites characteristic of subclass B3, with the amino acid motif His136, His138, His211, Asp140, His141, and His276. Among the six residues, His136, Asp140, and His211 exhibited the highest catalytic activity. We determined that CAR-2 can effectively hydrolyze cefalothin, cefuroxime and cefotaxime. The chloramphenicol resistance of the constructed E. coli DH5α strain was significantly reduced in the presence of CARR than in the absence of CARR. Compared with those for the wild-type P. diazotrophicus, MICs of cefalothin, cefuroxime and cefotaxime for the ΔCARR strain increased by 8-, 8- and 16-fold, respectively, and the expression level of blaCAR-2 also increased by approximately 10-fold.Conclusion: Overall, CAR-2 is a novel subclass B3 MBL of the CAR family that exhibits catalytic activity against cefalothin, cefuroxime and cefotaxime. CARR represses the expression of blaCAR-2, thereby reducing the resistance of P. diazotrophicus to these antibiotics. This provided a theoretical basis for revealing new mechanisms of pathogen resistance.
研究领域:
微生物学
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分子生物学
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感染性疾病
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抗生素耐药机制
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基因调控
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JOURNAL OF CONTROLLED RELEASE
·
2023
·
IF 10.8
Zhenmei Yang,Ying Liu,Kun Zhao,Weiqiang Jing,Lin Gao,Xianghui Dong,Yan Wang,Maosen Han,Chongdeng Shi,Chunwei Tang,Peng Sun,Rui Zhang,Zhipeng Fu,Jing Zhang,Danqing Zhu,Chen Chen,Xinyi Jiang
Hepatocellular carcinoma (HCC) is a prevalent and lethal disease, and tumor regression rarely occurs in advanced HCC patients due to limited effective therapies. Given the enrichment of macrophages in HCC and their role in tumor immunity, transforming them into chimeric antigen receptor macrophages (CAR-Ms) is thought to increase HCC cell-directed phagocytosis and tumoricidal immunity. To test this hypothesis, mRNA encoding CAR is encapsulated in a lipid nanoparticle (LNP) that targets liver macrophages. Notably, the LNPs adsorb specific plasma proteins that enable them to target HCC-associated macrophages. Moreover, mRNA encoding Siglec-G lacking ITIMs (Siglec-GΔITIMs) is codelivered to liver macrophages by LNP to relieve CD24-mediated CAR-Ms immune suppression. Mice treated with LNPs generating CAR-Ms as well as CD24-Siglec-G blockade significantly elevate the phagocytic function of liver macrophages, reduce tumor burden and increase survival time in an HCC mouse model. Arguably, our work suggests an efficacious and flexible strategy for the treatment of HCC and warrants further rigorous evaluation in clinical trials .
研究领域:
肿瘤学
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免疫治疗
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分子生物学
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纳米技术
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高影响因子
JOURNAL OF BIOMEDICAL SCIENCE
·
2023
·Vol. 30
·期 1
·1-19
·
IF 11
Li Peisi,Zhou Dawang,Chen Dongwen,Cheng Yikan,Chen Yuan,Lin Zhensen,Zhang Xi,Huang Zhihong,Cai Jiawei,Huang Wenfeng,Lin Yanyun,Ke Haoxian,Long Jiahui,Zou Yifeng,Ye Shubiao,Lan Ping
Background A large proportion of the patients with cancer do not respond to immunotherapies. Recent studies suggested an important role for tumor-infiltrating cytotoxic T lymphocytes (CTL) in enhancing response to immunotherapy. Here, we aim to identify gene that induce proliferative and cytotoxic states of CD8 + T cells, and to investigate its effect on CAR-T cells against colorectal cancer. Methods Correlation between the expression of IFI35 with the activation and cytotoxicity of CD8 + T cells was assessed with TCGA and proteomic databases. Then we constructed murine colon cancer cells over-expressing IFI35 and tested their effect on anti-tumor immunity in both immunodeficient and immunocompetent mouse models. Flow cytometry and immunohistochemistry were performed to assess the immune microenvironment. Western blot analysis was used to identify the potential down-stream signaling pathway regulated by IFI35. We further investigated the efficacy of the rhIFI35 protein in combination with immunotherapeutic treatment. Results The transcriptional and proteomic analysis of the activation and cytotoxicity of CD8 + T cells in human cancer samples demonstrated that IFI35 expression is correlated with increased CD8 + T cell infiltration and predicted a better outcome in colorectal cancer. The number and cytotoxicity of CD8 + T cells were significantly increased in IFI35-overexpressing tumors. Mechanistically, we identified that the IFNγ-STAT1-IRF7 axis stimulated IFI35 expression, and that IFI35-mediated regulation of CD8 + T cell proliferation and cytotoxicity was dependent on PI3K/AKT/mTOR signaling pathway in vitro. Furthermore, IFI35 protein enhanced the efficacy of CAR-T cells against colorectal cancer cells. Conclusion Our findings identify IFI35 as a new biomarker that can enhance the proliferation and function of CD8 + T cells, as well as increase the efficacy of CAR-T cells against colorectal cancer cells. Graphical Abstract
研究领域:
肿瘤学
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免疫学
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分子生物学
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癌症免疫疗法
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JOURNAL OF CONTROLLED RELEASE
·
2026
·
IF 12.4
Xiu Zhao,Kaikai Zhao,Mengfei Yi,Ruohan Yang,Xuyang Cui,Yi Zhang,Feng Li,Junjie Liu
Tumor chemokines that drives abnormal immune cell niches play a crucial role in impairing the chimeric antigen receptor (CAR)-T cell therapy against solid tumors. Here we develop a biomimetic nanoformulation with chemokine “leverage regulation” capabilities, which can establish matching migration signals to creat an immune cell niche that supports CAR-T cell therapy. The system is constructed using celecoxib (CXB)-loaded poly(d, l -lactide- co -glycolide) (PLGA) nanoparticles and subsequently camouflaging them with mesenchymal stem cell membranes with high CXCR4 expression. Leveraging the characteristic of tumors abnormally upregulating the chemokine CXCL12 to recruit CXCR4-positive immunosuppressive cells, the biomimetic nanoformulations can achieve efficient accumulation in the tumor tissue. These nanoformulations remodel the tumor chemokine milieu by upregulating immunogenic chemokines (CXCL9, CXCL10) to selectively recruit CAR-T cells, while simultaneously serving as a nanobait to neutralize CXCL12, thereby reducing the influx of immunosuppressive cells (such as CAR-Tregs and MDSCs) to enhance the killing efficacy of CAR-T cells. This dual modulation of the chemokine network significantly improves the therapeutic efficacy of CAR-T cells against solid tumors. Our approach represents a promising strategy for advancing CAR-T cell therapy toward clinical applications for soild tumors.
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Frontiers in Immunology
·
2026
·
IF 7
Gaohua Li,Lingping Zhao,Jingzhi Shen,Haiyang Wang,Jinru He,Yiming Liu,Ruge Zang,Tiantian Cui,Quan Zeng,Junnian Zhou,Wen Yue,Yanan Wang,Jiafei Xi
BackgroundCarcinoembryonic antigen (CEA) is overexpressed in most colorectal cancers (CRC) and serves as a promising immunotherapeutic target. However, current immune cell therapies against CRC show limited clinical efficacy, and the lack of renewable, standardized cell sources hampers broader application. The present study aimed to establish a scalable source of CEA-targeted natural killer (NK) cells from engineered human embryonic stem cells (hESCs).MethodshESCs were genetically engineered to express a CEA‑specific chimeric antigen receptor (CAR). The modified hESCs were subsequently differentiated into NK cells, yielding a homogeneous population of CAR hESC-NK cells. In vitro cytotoxicity against CEA-high CRC cell lines was evaluated using LDH and IFN-γ release assays. For in vivo assessment, we established a xenograft mouse model bearing CEA-positive CRC tumors. Following adoptive transfer of CAR-hESC-NK cells, tumor growth was monitored by bioluminescence imaging, overall survival was recorded for Kaplan–Meier analysis, and the persistence of transferred cells in peripheral blood was examined by flow cytometric analysis at serial time points.ResultsIn vitro, CAR hESC‑NK cells demonstrated potent and specific cytotoxicity against CEA-high CRC cell lines, along with significantly enhanced cytokine secretion compared to unmodified NK cells. In the xenograft model, adoptive transfer of CAR hESC‑NK cells resulted in marked tumor growth inhibition and prolonged survival, with no observable off‑target toxicity. Importantly, the CAR CEA‑ESC line maintained stable transgene expression and could serve as a renewable seed cell source.ConclusionWe have established a scalable platform for producing CEA‑targeted NK cells from engineered hESCs, providing an off‑the‑shelf, homogeneous cell product with robust antitumor activity. This strategy provides effective and specific immunotherapy against CEA-positive colorectal cancer and holds potential for other CEA-expressing malignancies.
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SCIENCE OF THE TOTAL ENVIRONMENT
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2021
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IF 7.963
Yingfei Sheng,Yi Liu,Kewei Wang,James V. Cizdziel,Yichun Wu,Ying Zhou
As one of the most common and persistent emerging human-made pollutants, microplastics (MPs) have become a global environmental problem. Although many studies have demonstrated the impact of MPs on aquatic organisms, the potential effects on terrestrial fauna are relatively unknown, even though soil is a major sink for plastic pollution. In this study, earthworms ( Eisenia fetida ) were exposed to MPs from tires (TMPs) in artificial soil at different TMP concentrations (0, 1, 5, 10, and 20%, w /w), at different TMP size fractions (2 mm–350 μm; 350 μm–50 μm; 50 μm–25 μm; and <25 μm), and time periods (14 and 28 days). The purpose was to evaluate the impact of TMPs on the bioaccumulation of heavy metals (Zn, Cd, and Pb) and levels of oxidative stress in the earthworms. Fluorescence imaging of TMPs stained with Nile red dye clearly showed that the worms had ingested the TMPs, with smaller particles preferentially consumed. The ingested TMPs had altered the surface morphology, degradation that increased surface area and uptake of heavy metals into the organisms. Earthworms exposed to TMPs at the higher concentrations showed clear signs of oxidative stress, with significantly ( p < 0.05) increased catalase and peroxidase activity and lipid peroxidation levels, and reduced activity of superoxide dismutase and glutathione S-transferase. SOD, POD, and CAT are important cell protection enzymes that prevent excessive levels of reactive oxygen species in cells. MDA represents the total scavenging ability of free radicals in the tissue. SOD decreases and MDA accumulates rapidly. GST has a detoxification effect. Overall, this study adds to the growing body of evidence that TMPs are harmful to certain terrestrial organisms.
研究领域:
环境科学
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生态毒理学
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土壤生物学
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Journal of Cleaner Production
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2026
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IF 10.7
Yang Ren,Xinhong Yuan,Yang Hu,Hongyun Hu,Kai Jiang,Zhen Wu,Yongxin Duan,Xian Li,Hong Yao
Adaptable molten salt treatment yields similar products from car and truck tires. • Molten NaOH-Na 2 CO 3 enables in-situ upgrading of car tire pyrolysis products. • The absolute content of BTX in tire pyrolysis oil increased by 1.5-2.8 times. • Maximum ash and sulfur removal in tire pyrolysis char reached 71% and 65%. • The H 2 content in the tire pyrolysis gas increased by a maximum of 19 times. Waste passenger car tires present significant recycling challenges due to their low thermal conductivity and high ash and sulfur contents, which frequently result in suboptimal product quality during conventional pyrolysis. To overcome the limitations of complex post-treatment processes, a molten salt thermal treatment method utilizing NaOH-Na 2 CO 3 medium was proposed to achieve the in-situ upgrading of pyrolysis products. In this study, the effects of operating temperature (425-575 °C) and feedstock particle size (6-10 mm) on products were systematically evaluated. Experimental results indicate that the molten salt medium significantly enhanced heat transfer and catalytic cracking, which effectively fixed CO 2 and led to a 19-fold surge in H 2 production, peaking at 5.57 mmol/g. For the liquid-phase products, the introduction of molten salt significantly promoted the dealkylation of substituted aromatics, increasing the benzene concentration by 5.43 to 8.61 times. Furthermore, the molten salt effectively overcame the mass transfer resistance of larger particles, facilitating the extensive formation of monocyclic aromatic hydrocarbons. Notably, at the solid-phase interface, the molten salt acted as an efficient in-situ purifying agent, reacting directly with inorganic impurities (SiO 2 , CaCO 3 ) and sulfides to form highly water-soluble salts. This mechanism achieved a maximum deashing ratio of 71% and a desulfurization ratio of 36.74%, yielding a high-value pyrolytic char with a carbon content exceeding 90%. Overall, this approach demonstrates that the thermal conversion products of low-quality passenger car tires can be significantly upgraded, providing a highly efficient pathway for comprehensive resource recovery. Download: Download high-res image (371KB) Download: Download full-size image
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INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2026
·Vol. 27
·期 2
·607
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IF 4.9
Jiaxuan Zhao,Youyong Wang,Yixuan Wang,Ge Dong,Han Wu,Yeting Cui,Lixing Gu,Fenfang Zhao,Guanlin Zhao,Jinyu Kang,Qian Zhang,Nan Liu,Ning Wang,Xiao Sun,Yao Xu,Tongcun Zhang,Jiangzhou Shi
The manufacturing process contributes significantly to the proliferation, metabolic state, and functional persistence of chimeric antigen receptor (CAR)-T cells. However, how different culture systems regulate CAR-T cell metabolism and thereby influence their long-term antitumor activity remains poorly understood. In this study, we compared dynamic cultivation using a wave bioreactor with static expansion systems (gas-permeable and conventional T-flasks) for the production of CD99-specific CAR-T cells. CAR-T cells expanded by the wave bioreactor exhibited faster proliferation and stronger cytotoxicity during culture. Upon repeated antigen stimulation, they retained these enhanced functional properties and showed the reduced expression of immune checkpoint molecules, preferentially preserved memory-like subsets, and displayed transcriptional features consistent with memory maintenance and exhaustion resistance. Targeted metabolomic profiling revealed enhanced Tricarboxylic Acid (TCA) cycle activity and features consistent with sustained oxidative phosphorylation, supporting mitochondrial-centered metabolic reprogramming. In a Ewing sarcoma xenograft model, wave bioreactor-cultured CAR-T cells showed a greater percentage of memory-like tumor-infiltrating lymphocytes. Collectively, these results indicate that wave bioreactor-based dynamic cultivation promotes mitochondrial metabolic reprogramming, which is characterized by an enhanced TCA cycle and sustained oxidative phosphorylation, thereby sustaining CAR-T cell functionality and providing a robust platform for the manufacturing of potent and durable cellular therapeutics.
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MOLECULES
·
2012
·Vol. 17
·期 9
·10994-11009
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IF 2.386
Yunxia Zhang,Yunlan Li,Qingshan Li
The specific aims of this study were to evaluate the inhibition effect on CYP3A of di-n-butyl-di-(4-chlorobenzohydroxamato)tin (IV) (DBDCT), a tin-based complex with high antitumor activity, and the probable mechanism(s) of this action. Adult male SD rats were treated separately with natural saline (NS), lipopolysaccharide (LPS, 5 mg/kg), DBDCT (1.25, 2.5 and 5.0 mg/kg) intraperitoneally for 2 days after induction of CYP3A with dexamethasone (DEX, 100 mg/kg) for 4 days. Western blot analysis and fluorescent quantitation PCR (FQ-PCR) were conducted to determine the changes in expression of CYP3A, PXR, CAR and RXR. The biological accumulation of DBDCT and total Sn were determined by high-performance liquid chromatography (HPLC) and atomic fluorescence spectrometry (AFS). CYP450 content and CYP3A activities were significantly inhibited (p< 0.05) in DBDCT-treated rats compared with the control group, as was the expression of CYP3A (p< 0.05) at both protein and mRNA levels. In DBDCT-treated groups, the expression of PXR protein and mRNA increased, while the expression of CAR decreased. The biological accumulation of DBDCT and Sn in rat livers treated with DBDCT was high. The accumulation of DBDCT and Sn due to the inhibition of CYP3A may be involved in the mechanism of toxicity of DBDCT in rat liver.Keywords:cytochrome P450 3A (CYP3A);di-n-butyl-di-(4-chlorobenzohydroxamato)tin (IV) (DBDCT);inhibition;rat
研究领域:
药理学
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毒理学
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分子生物学
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Journal of Energy Chemistry
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2023
·
IF 13.1
Baoyu Wang,Peng Zhou,Ximing Yan,Hu Li,Hongguo Wu,Zehui Zhang
In this work, a dual-size MOF-derived Co catalyst (0.2Co 1-NPs @NC) composed of single atoms (Co 1 ) and highly dispersed nanoparticles (Co NPs) was prepared by in-situ Zn evaporation for the high-performance conversion of lignin-derived o- methoxyphenols (lignin oil) to cyclohexanols (up to 97% yield) via cascade demethoxylation and dearomatization. Theoretical calculations elaborated that the dual-size Co catalyst exhibited a cooperative effect in the selective demethoxylation process, in which the Co NPs could initially dissociate hydrogen at lower energies while Co 1 remarkably facilitated the cleavage of the C Ar − OCH 3 bond. Moreover, the intramolecular hydrogen bonds formed in the o -methoxy-containing phenols were found to result in a decrease in the bond energy of the C Ar − OCH 3 bond, which was more prone to be activated by the dual-size Co sites. Notably, the pre-hydrogenated intermediate (e.g., 2-methoxycyclohexanol from guaiacol) is difficult to undergo demethoxylation, indicating that the selective C Ar − OCH 3 bond cleavage is a prerequisite for the synthesis of cyclohexanols. The 0.2Co 1-NPs @NC catalyst was highly recyclable with a neglect decline in activity during five consecutive cycles. This cooperative catalytic strategy based on the metal size effect opens new avenues for biomass upgrading via enhanced C O bond cleavage of high selectivity.
研究领域:
化学工程
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催化科学
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Advanced Science
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2025
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IF 14.1
Renjie Cao,Tingting Zhao,Ying Wang,Shaofei Song,Yuan Li,Mengling Hou,Yanxin Zhang,Sengpeng Wong,Siqi Wang,Yuran Wang,Hong Peng,Min Huang,Yiming Jiang
Macrophage proinflammatory hyperactivation drives the pathogenesis of acute liver injury, a common complication of sepsis. The role of the nuclear receptor constitutive androstane receptor (CAR) in endotoxin-induced liver injury remains unclear. Here, this study reports that CAR is highly expressed in human and murine macrophages. CAR activation markedly attenuated endotoxin-induced liver damage, alleviating hepatocyte death and hepatic inflammation. Macrophage-hepatocyte coculture confirmed that CAR inhibited inflammation through macrophage crosstalk. CAR-mediated hepatoprotection and anti-inflammatory effects are absent in AAV8- F4/80- sh Car -treated mice, confirming the essential role of CAR in macrophages. Mechanistically, CAR is found to interact with Tlr4 , and the suppressive effects of CAR on TLR4 are proven in Tlr4 −/− mice. Furthermore, CAR activation reduced LPS-induced inflammation in hMDMs, BMDMs, RAW264.7, and THP-1 cells, and Car or Tlr4 knockdown abolished CAR-mediated immunosuppression. Overall, these findings showed that macrophage CAR activation attenuated endotoxin-induced liver injury and hepatic inflammation through the TLR4 signaling pathway, providing insights for treating inflammatory liver diseases.
研究领域:
免疫学
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分子生物学
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肝病学
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药理学
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细胞信号传导
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JOURNAL OF CONTROLLED RELEASE
·
2026
·
IF 12.4
Nan Yang,Linlin Tao,Xiaoxiao Hu,Chengqian Shang,Nian Huang,Qiaoqiong Lin,Fangnan Lv,Zhiyong Sun,Chengcheng Gu,Yunfei Lian,Wei Xu,Meirong Huo
CAR-T therapy holds immense potential for eradicating residual tumors and restoring postoperative immune surveillance. However, nutrient deprivation and impaired immune contact within the postoperative tumor microenvironment (PTME) severely hinder the therapeutic efficacy of CAR-T cells. Herein, we developed a self-reinforcing hydrogel system that dually modulated metabolism and immune contact to reprogram the PTME. Specifically, the c-Myc inhibitor JQ1 and bovine serum albumin (BSA)-decorated nanoparticles loaded with the actin cytoskeleton inhibitor NP-G2–044 (BPG) were encapsulated into a reactive oxygen species (ROS)-sensitive hydrogel (JQ1 + BPG@gel). Upon implantation into the surgical cavity, JQ1 + BPG@gel underwent ROS-responsive degradation, continuously releasing agents. JQ1 not only suppressed glycolysis in tumor cells, thereby redirecting nutrients toward CAR-T cells and alleviating lactic acid-induced immunosuppression, but also upregulated the gradients of chemokines CCL5 and CXCL10, thus providing a strong driving force for CAR-T cell infiltration. Notably, JQ1-induced glycolysis inhibition significantly enhanced the tumor uptake of BSA, establishing a self-reinforcing internalization process for BPG. The increased intracellular accumulation of BPG remodeled actin cytoskeleton, enhancing the spatial co-localization between T cells and tumor cells while reducing the steric hindrance imposed by complex cellular topology, thereby facilitating the formation of immune contact and tumor lysis. This hydrogel system improved the anti-recurrence efficacy of CAR-T therapy by 9-fold compared with monotherapy while establishing durable immune surveillance via memory T-cell differentiation within lymphoid organs. This research offers an innovative strategy to improve postoperative tumor cure rates and potentiate CAR-T immunotherapy.
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