VIROLOGY
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2023
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IF 3.7
Xiaoping Huang,Xiao Wang,Yaqi Sun,Ling Li,Anna Li,Wentao Xu,Xiaolan Xie,Yong Diao
Because it is safe and has a simple genome, recombinant adeno-associated virus (rAAV) is an extremely appealing vector for delivery in in vivo gene therapy. However, its low transduction efficiency for some cells, limits its further application in the field of gene therapy. Bleomycin is a chemotherapeutic agent approved by the FDA whose effect on rAAV transduction has not been studied. In this study, we systematically investigated the effect of Bleomycin on the second-strand synthesis and used CRISPR/CAS9 and RNAi methods to understand the effects of Bleomycin on rAAV vector transduction, particularly the effect of DNA repair enzymes. The results showed that Bleomycin could promote rAAV2 transduction both in vivo and in vitro. Increased transduction was discovered to be a direct result of decreased cytoplasmic rAAV particle degradation and increased second-strand synthesis. TDP1 , PNKP , and SETMAR are required to repair the DNA damage gap caused by Bleomycin, TDP1, PNKP, and SETMAR promote rAAV second-strand synthesis. Bleomycin induced DNA-PKcs phosphorylation and phosphorylated DNA-PKcs and Artemis promoted second-strand synthesis. The current study identifies an effective method for increasing the capability and scope of in-vivo and in-vitro rAAV applications, which can amplify cell transduction at Bleomycin concentrations. It also supplies information on combining tumor gene therapy with chemotherapy.
研究领域:
分子生物学
·
基因治疗
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癌症研究
·
药理学
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DOI
PubMed
高影响因子
LIFE SCIENCES
·
2024
·
IF 5.2
Yi Xiao,Jiahe Zhang,Xinran Li,Pinxuan Liu,Bo Gou,Zeyu Gao,Moshi Song
Background The role of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is multifaceted, paradoxically promoting both cell survival and cell death across multiple organs. However, its impact on lung homeostasis remains elusive. Here, we investigate the function of DNA-PKcs in mouse lungs, aiming to elucidate its role for lung abnormalities associated with DNA-PKcs deficiency. Materials and methods Histological assessment and immunohistochemistry were used to reveal the pathological changes of the lungs in DNA-PKcs-deficient mice. Transcriptomic analysis identified differentially expressed genes and pathways in DNA-PKcs-deficient lungs. Furthermore, mitochondrial dysfunction induced by DNA-PKcs deficiency was investigated by qPCR and immunoblotting. Mouse primary lung fibroblasts were used to evaluate the potential therapeutic effect of inhibiting mitochondrial fission with Mdivi-1. Key findings In DNA-PKcs-deficient mouse lungs, we observed pathological changes including alveolar septal thickening, capillary congestion and hemorrhage, along with lung cell proliferation. Transcriptome analysis revealed an upregulation of the reactive oxygen species (ROS) biosynthesis process and the apoptotic signaling pathway caused by DNA-PKcs deficiency. Further investigations demonstrated that DNA-PKcs deficiency led to mitochondrial dysfunction and increased oxidative stress, along with increased cell apoptosis in the mouse lungs. Notably, we detected enhanced phosphorylation of the mitochondrial fission protein DRP1 in DNA-PKcs-deficient mouse lungs. Intriguingly, inhibiting mitochondrial fission using Mdivi-1 suppressed cell death in primary mouse lung fibroblasts with siRNA-mediated DNA-PKcs knockdown. Significance Our study provides insights into the crucial role of DNA-PKcs in sustaining lung homeostasis via the maintenance of mitochondrial functionality and provides a therapeutic strategy targeting mitochondrial fission against DNA-PKcs deficiency-associated lung diseases.
研究领域:
分子生物学
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细胞生物学
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肺生理学
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病理学
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产品关联
DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF MEDICAL RESEARCH
·
2025
·
IF 3.4
Yun Chen,Yang Wang,Shuai Shen,Libin Zhang,Jun Liu,Xiangyun Yan,Hao Peng,Zheyuan Xu
RHDBB1 could arrest cell cycle in the TP53 mutant NCI-H596 cells; RHDBB1 promotes NSCLS by reducing DNA-PKcs dependent cells apoptosis; Targeting both TP53 and RHDBB1 may have beneficial effects on NSCLC.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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癌症研究
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR STRUCTURE
·
2026
·
IF 4.9
Junjie Tong,Tong Wang,Xixi Wang,Jingying Chen,Changbin Yu,Yanhui Fu,Miaoshan Lu
AI-predicted and experimental structures were cross-checked for screening. • Structure-source effects on ligand enrichment were target-dependent. • Multiple DNA-PKcs structures prioritized partly distinct candidate sets. • L-365,260 and Fmoc-Tic-OH showed apparent micromolar DNA-PKcs binding. • Both compounds produced radiosensitizing phenotypes in NSCLC cells. Radioresistance remains a major challenge in non-small-cell lung cancer (NSCLC) radiotherapy, motivating the search for compounds that enhance tumor-cell responses to irradiation. Artificial intelligence (AI)-predicted structures expand access to structure-based virtual screening, but how they compare with and complement experimentally determined structures remains unclear. Here, we compared experimental, AlphaFold2, AlphaFold3 and RoseTTAFold All-Atom structures of four DNA damage-response proteins—ATM, ATR, DNA-PKcs and PARP1—using structural analyses and retrospective active-decoy enrichment. AI-predicted and experimental structures of DNA-PKcs were then used to screen 79,263 natural-product-related compounds from ZINC20. The prospective screen yielded 13 filtered candidates and showed that the three DNA-PKcs structures prioritized partially overlapping chemical sets. Docking, ADMET prediction, molecular dynamics and molecular mechanics/Poisson–Boltzmann surface area (MM/PBSA) calculations were used to rank candidates, and L-365,260 (ZINC000003797417) and Fmoc-Tic-OH (ZINC000000621966) were selected for experimental characterization. Surface plasmon resonance yielded apparent steady-state DNA-PKcs dissociation constant (KD) estimates of 11 and 24 μM for L-365,260 and Fmoc-Tic-OH, respectively. In A549 and H1299 cells, the compounds produced model-derived sensitizer enhancement ratios at 50% survival (SER50) ranging from 1.87 to 2.17. Proteomic profiling, apoptosis, cell-cycle and γH2AX analyses characterized treatment-associated cellular responses. Within this DNA-PKcs case study, the results indicate that AI-predicted and experimental structures can provide complementary views of a binding site and prioritize partially non-overlapping candidate sets. This comparative workflow offers a practical framework for cross-checking structural sources and prioritizing compounds in structure-based screening. Download: Download high-res image (212KB) Download: Download full-size image
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高影响因子
EUROPEAN JOURNAL OF MEDICAL RESEARCH
·
2025
·Vol. 30
·期 1
·1-13
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IF 3.4
Luo Liling,Jiang Chao,Xie Songxi
Background Previous studies showed that high mobility group box-1 (HMGB1) facilitates the initiation and progression of esophageal squamous cell carcinoma (ESCC), and the current research investigated the detailed mechanisms implicated.Methods The impact of HMGB1 and IGFBP3 levels on the survival of ESCC was examined by plotting Kaplan–Meier (KM) curves based on the data collected from The Cancer Genome Atlas (TCGA). Quantitative real-time PCR (qRT-PCR) was performed to detect the expressions of HMGB1 in both human esophageal epithelial cells (HEEC) and ESCC cells. After cell transfection, the proliferation of ESCC cells was measured, and the cell metastasis was determined based on the levels of cadherins (CDHs) and Vimentin (VIM). Macrophage polarization was determined by calculating the mean fluorescence intensity (MFI) of CD206 and CD86. In addition, co-immunoprecipitation and immunoblotting were applied to evaluate the interaction between insulin-like growth factor binding protein 3 (IGFBP3)/DNA-dependent protein kinase catalytic subunit (DNA–PKcs) and HMGB1.Results A high level of HMGB1 was predictive of an unfavorable prognosis of ESCC (p < 0.05). HMGB1 showed a higher expression in ESCC cells (p < 0.05), while knockdown of HMGB1 inhibited ESCC cell proliferation, downregulated the levels of CDH2 and VIM and upregulated the level of CDH1 (p < 0.05). In contrast, overexpressed HMGB1 showed the opposite effects (p < 0.05), suggesting the role of HMGB1 in the epithelial–mesenchymal transition (EMT) of ESCC. After the knockout of HMGB1, the MFI of CD86 was increased but that of CD206 was reduced, indicating the polarization towards M1 macrophages (p < 0.05). However, the results were reversed when HMGB1 was overexpressed (p < 0.05). Meanwhile, HMGB1 could interact with the IGFBP3/DNA–PKcs complex (p < 0.05). Low-expressed IGFBP3 was predictive of an unfavorable prognosis of ESCC, and IGFBP3 silencing promoted the proliferation of ESCC cells (p < 0.05). Besides, HMGB1 and IGFBP3 could act antagonistically in influencing the proliferation of ESCC cells and macrophage polarization.Conclusion sThrough in vitro experiments, this study found that HMGB1 was linked to the proliferation and polarization of macrophages in ESCC, providing novel evidence for the role of HMGB1 in ESCC development.
研究领域:
肿瘤学
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分子生物学
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免疫学
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癌症研究
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产品关联
DOI
PubMed
高影响因子
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