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Targeting Orthotopic Glioma in Mice with Genetically Engineered Salmonella typhimurium
Wen, Min,Jung, Shin,Moon, Kyung-Sub,Jiang, Shen Nan,Li, Song-Yuan,Min, Jung-Joon The Korean Neurosurgical Society 2014 Journal of Korean neurosurgical society Vol.55 No.3
Objective : With the growing interests of bacteria as a targeting vector for cancer treatment, diverse genetically engineered Salmonella has been reported to be capable of targeting primary or metastatic tumor regions after intravenous injection into mouse tumor models. The purpose of this study was to investigate the capability of the genetically engineered Salmonella typhimurium (S. typhimurium) to access the glioma xenograft, which was monitored in mouse brain tumor models using optical bioluminescence imaging technique. Methods : U87 malignant glioma cells (U87-MG) stably transfected with firefly luciferase (Fluc) were implanted into BALB/cAnN nude mice by stereotactic injection into the striatum. After tumor formation, attenuated S. typhimurium expressing bacterial luciferase (Lux) was injected into the tail vein. Bioluminescence signals from transfected cells or bacteria were monitored using a cooled charge-coupled device camera to identify the tumor location or to trace the bacterial migration. Immunofluorescence staining was also performed in frozen sections of mouse glioma xenograft. Results : The injected S. typhimurium exclusively localized in the glioma xenograft region of U87-MG-bearing mouse. Immunofluorescence staining also demonstrated the accumulation of S. typhimurium in the brain tumors. Conclusion : The present study demonstrated that S. typhimurium can target glioma xenograft, and may provide a potentially therapeutic probe for glioma.
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Wen, Min,Zheng, Jin Hai,Choi, Jin Myung,Pei, Jian,Li, Chun-Hao,Li, Song-Yuan,Kim, In-Young,Lim, Sa-Hoe,Jung, Tae-Young,Moon, Kyung-Sub,Min, Jung-Joon,Jung, Shin Elsevier 2018 Cancer letters Vol.433 No.-
<P><B>Abstract</B></P> <P>Glioma is one of the most devastating and refractory cancers. The main factors underlying therapeutic failure include extremely invasive characteristics and lack of effective methods for drug delivery. Attenuated <I>Salmonella</I> strains presented a high concentration of tumor targets in various types of cancer models, suggesting a role as potential vectors for drug delivery. In this study, we genetically engineered an attenuated strain of <I>Salmonella</I> as an anti-invasive vector for the targeted delivery and expression of tissue inhibitor of metalloproteinases 2 (TIMP-2) in an orthotopic nude mouse model of glioma. The bioluminescence signals related to tumor size significantly declined in the TIMP-2-expressing <I>Salmonella</I> (SLpTIMP-2)-treated group compared with the control group. Compared with the control group with a survival rate of an average of 33 days, the SLpTIMP-2 group showed an extended survival rate by nearly 60% and lasted an average period of 53 days with TIMP-2 induction. These results indicated the promising therapeutic potential of <I>S. typhimurium</I> for targeted delivery and secretion of TIMP-2 in glioma.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Intracranial injection of <I>Salmonella</I> has been demonstrated to be a more effective than tail vein injection. </LI> <LI> Treatment with TIMP-2-expressing bacteria showed down regulation of MMP-2 in orthotopic glioma. </LI> <LI> TIMP-2-expressing bacteria significantly inhibited tumor growth and elongated animal survive. </LI> </UL> </P>
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Yuan, Dong-Mei,Li, Qian,Zhang, Qin,Xiao, Xin-Wu,Yao, Yan-Wen,Zhang, Yan,Lv, Yan-Ling,Liu, Hong-Bin,Lv, Tang-Feng,Song, Yong Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.4
Objectives: Can addition of neurokinin-1 receptor antagonists (NK1-RAs) be considered as an ideal strategy for the prevention of chemotherapy-induced nausea and vomiting (CINV)? Researchers differ on this question. Materials and Methods: Electronic databases were searched for randomized control trials (RCTs) that evaluated the effectiveness and safety of NK1-RAs in preventing CINV. The primary end point was complete response (CR) in the acute, delayed, and overall phases after chemotherapy. Subgroup analyses evaluated the types of NK1-RAs, routines of administration, types of malignancies, regimens used in combination with NK1-RAs, and age of patients included in the studies. The incidences of different types of adverse events were also extracted to estimate the safety of NK1-RAs. Results: A total of 38 RCTs involving 13,923 patients were identified. The CR rate of patients receiving NK-RAs was significantly higher than patients in the control groups during overall phase (70.8% vs 56.0%, P<0.001), acute phase (85.1% vs 79.6%, P<0.001), and delayed phase (71.4% vs 58.2%, P<0.001). There were three studies including patients of children or adolescents, the CR rate was also significantly higher in the treatment group (overall phase: OR=2.807, P<0.001; acute phase: OR=2.863, P =0.012; delayed phase: OR=2.417, P<0.001). For all the other outcomes, patients in the NK1-RAs groups showed improvements compared to the control groups (incidence of nausea: 45.2% vs 45.9%, P<0.001; occurrence of vomiting: 22.6% vs 38.9%, P<0.001; usage of rescue drugs: 23.5% vs 34.1%, P<0.001). The pooled side effects from NK1-RAs did not significantly differ from previous reports and the toxicity rates in patients less than eighteen years old also did not diff between the two groups (P=0.497). However, we found that constipation and insomnia were more common in the patients of control groups, whereas diarrhea and hiccups were more frequently detected in patients receiving NK1-RAs. Conclusions: NK1-RAs improved the CR rate of CINV. They are effective for both adults and children. The use of NK1-RAs might be associated with the appearance of diarrhea and hiccups, while decreasing the possibility of constipation and insomnia.
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Xue Song,Jing Guo,Wen-xiu Ma,Zhi-yuan Ji,Li-fang Zou,Gong-you Chen,Hua-song Zou 한국미생물학회 2015 The journal of microbiology Vol.53 No.5
To identify novel virulence genes, a mutant library of Xanthomonas citri subsp. citri 29-1 was produced using EZ-Tn5 transposon and the mutants were inoculated into susceptible grapefruit. Forty mutants with altered virulence phenotypes were identified. Nine of the mutants showed a complete loss of citrus canker induction, and the other 31 mutants resulted in attenuated canker symptoms. Southern blot analysis revealed that each of the mutants carried a single copy of Tn5. The flanking sequence was identified by plasmid rescue and 18 different ORFs were identified in the genome sequence. Of these 18 ORFs, seven had not been previously associated with the virulence of X. citri subsp. citri and were therefore confirmed by complementation analysis. Real-time PCR analysis showed that the seven genes were upregulated when the bacteria were grown in citrus plants, suggesting that the expression of these genes was essential for canker development.
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Wei Yuan,Qing-feng Wang,Wen-han Pei,Si-yu Li,Tian-min Wang,Hui-peng Song,Dan Teng,Ting-guo Kang,Hui Zhang 고려인삼학회 2024 Journal of Ginseng Research Vol.48 No.1
Background: Ginseng (Panax ginseng Mayer) is an important natural medicine. However, a long culture period andchallenging quality control requirements limit its further use. Although artificial cultivation can yield a sustainablemedicinal supply, research on the association between the transplantation and chaining of metabolicnetworks, especially the regulation of ginsenoside biosynthetic pathways, is limited. Methods: Herein, we performed Liquid chromatography tandem mass spectrometry based metabolomic measurementsto evaluate ginsenoside accumulation and categorise differentially abundant metabolites (DAMs). Transcriptome measurements using an Illumina Platform were then conducted to probe the landscape of geneticalterations in ginseng at various ages in transplantation mode. Using pathway data and crosstalk DAMs obtainedby MapMan, we constructed a metabolic profile of transplantation Ginseng. Results: Accumulation of active ingredients was not obvious during the first 4 years (in the field), but followingtransplantation, the ginsenoside content increased significantly from 6 8 years (in the wild). Glycerolipidmetabolism and Glycerophospholipid metabolism were the most significant metabolic pathways, as Lipids andlipid-like molecule affected the yield of ginsenosides. Starch and sucrose were the most active metabolic pathwaysduring transplantation Ginseng growth. Conclusion: This study expands our understanding of metabolic network features and the accumulation of specificcompounds during different growth stages of this perennial herbaceous plant when growing in transplantationmode. The findings provide a basis for selecting the optimal transplanting time.
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Yan-Qing Duan,Song-Tao He,Qing-Qing Li,Ming-Feng Wang,Wen-Yuan Wang,Wei Zhe,Yong-Hong Cao,Ming-He Mo,Yu-Long Zhai,Wen-Jun Li 한국미생물학회 2013 The journal of microbiology Vol.51 No.3
A Gram-positive, catalase- and oxidase-positive, strictly aerobic, endospore-forming rod bacterium, designated K3514T, was isolated from the leaves of Nicotiana tabacum. The strain was able to grow at temperatures of 8–40°C, pH 5.0–10.0 and NaCl concentrations of 0–7%. The predominant quinones (>30%) of this strain were MK-7(H2) and MK-7. Phylogenetic analysis of 16S rRNA gene sequence showed that strain K3514T was affiliated to the genus Lysinibacillus, with its closest relatives being Lysinibacillus mangiferihumi (98.3% sequence similarity), Lysinibacillus sphaericus (97.9% sequence similarity), Lysinibacillus fusiformis (97.4% sequence similarity), and Lysinibacillus xylanilyticus (97.3% sequence similarity). However, low levels of DNA-DNA relatedness values suggested that the isolate was distinct from the other closest Lysinibacillus species. Additionally, based on analysis of morphological, physiological, and biochemical characteristics, the isolate could be differentiated from the closest known relatives. Therefore, based on polyphasic taxonomic data, the novel isolate likely represents a novel species, for which the name Lysinibacillus tabacifolii sp. nov. and the type strain K3514T (=KCTC 33042T =CCTCC AB 2012050T) are proposed.
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Qian Chen,Jun Song,Wen‑Ping Du,Li‑Yuan Xu,Yun Jiang,Jie Zhang,Xiao‑Li Xiang,Gui‑Rong Yu 한국유전학회 2018 Genes & Genomics Vol.40 No.10
Semi-dwarfism is an agronomically important trait in breeding for stable high yields and for resistance to damage by wind and rain (lodging resistance). Many QTLs and genes causing dwarf phenotype have been found in maize. However, because of the yield loss associated with these QTLs and genes, they have been difficult to use in breeding for dwarf stature in maize. Therefore, it is important to find the new dwarfing genes or materials without undesirable characters. The objectives of this study were: (1) to figure out the inheritance of semi-dwarfism in mutants; (2) mapping dwarfing gene or QTL. Maize inbred lines ‘18599’ and ‘DM173’, which is the dwarf mutant derived from the maize inbred line ‘173’ through 60Co-γ ray irradiation. F2 and BC1F1 population were used for genetic analysis. Whole genome resequencing-based technology (QTL-seq) were performed to map dwarfing gene and figured out the SNP markers in predicted region using dwarf bulk and tall bulk from F2 population. Based on the polymorphic SNP markers from QTL-seq, we were fine-mapping the dwarfing gene using F2 population. In F2 population, 398 were dwarf plants and 135 were tall plants. Results of χ2 tests indicated that the ratio of dwarf plants to tall plants was fitted to 3:1 ratio. Furthermore, the χ2 tests of BC1F1 population showed that the ratio was fitted to 1:1 ratio. Based on QTL-seq, the dwarfing gene was located at the region from 111.07 to 124.56 Mb of chromosome 9, and we named it rht-DM. Using traditional QTL mapping with SNP markers, the rht-DM was narrowed down to 400 kb region between SNP-21 and SNP-24. The two SNPs were located at 0.43 and 0.11 cM. Segregation analysis of F2 and BC1F1 indicated that the dwarfing gene was likely a dominant gene. This dwarfing gene was located in the region between 115.02 and 115.42 Mb on chromosome 9.
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Jian Son,Hong-Li Wang,Ke-Han Song,Zhi-Wen Ding,Hai-Lian Wang,Xiao-Sheng Ma,Fei-Zhou Lu,Xin-Lei Xia,Ying-Wei Wang,Fei-Zou,Jian-Yuan Jiang 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-
This study was carried out to explore the roles of circular RNAs (circRNAs) in nucleus pulposus (NP) tissues in intervertebral disc degeneration (IDD). Differentially expressed circRNAs in IDD and normal NP tissues were identified based on the results of microarray analysis. Bioinformatics techniques were employed to predict the direct interactions of selected circRNAs, microRNAs (miR), and mRNAs. CircRNA_104670 was selected as the target circRNA due to its large multiplier expression in IDD tissues. After luciferase reporter and EGFP/RFP reporter assays, we confirmed that circRNA_104670 directly bound to miR-17-3p, while MMP-2 was the direct target of miR-17-3p. The receiver-operating characteristic (ROC) curve showed that circRNA_104670 and miR-17-3p had good diagnostic significance for IDD (AUC circRNA_104670 = 0.96; AUC miRNA-17-3p = 0.91). A significant correlation was detected between the Pfirrmann grade and expression of circRNA_104670 (r = 0.63; p = 0.00) and miR-17-3p (r = −0.62; p = 0.00). Flow-cytometric analysis and the MTT assay showed that interfering with circRNA_104670 using small interfering RNA (siRNA) inhibited NP cell apoptosis (p < 0.01), and this inhibition was reduced by interfering with miR-17-3p. Interfering with circRNA_104670 suppressed MMP-2 expression and increased extracellular matrix (ECM) formation, which were also reduced by interfering with miR-17-3p. Finally, an MRI evaluation showed that circRNA_104670 inhibition mice had a lower IDD grade compared with control mice (p < 0.01), whereas circRNA_104670 and miRNA-17-3p inhibition mice had a higher IDD grade compared with circRNA_104670 inhibition mice (p < 0.05). CircRNA_104670 is highly expressed in the NP tissues of IDD and acts as a ceRNA during NP degradation.
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