A hypoxia-regulated adeno-associated virus vector for cancer-specific gene therapy Neoplasia 2001 3: 255–263, Gorski DH et al.
Levels of EGFP expression were assessed after 7 h reoxygenation. Census and consensus in bacterial ecosystems: The LuxR-LuxI family of quorum-sensing transcriptional regulators, Extensive and specific responses of a eukaryote to bacterial quorum-sensing signals, Bacteria-host communication: The language of hormones, Structure of the autoinducer required for expression of, The acyl-homoserine lactone-type quorum-sensing system modulates cell motility and virulence of, Indole-3-acetic acid in microbial and microorganism-plant signaling, Auxin: Regulation, action, and interaction, Ambewelamides A and B, antineoplastic epidithiopiperazinediones isolated from the lichen, Cyclodipeptide synthases are a family of tRNA-dependent peptide bond-forming enzymes, Aux/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements, The F-box protein TIR1 is an auxin receptor, The Arabidopsis F-box protein TIR1 is an auxin receptor, Common virulence factors for bacterial pathogenicity in plants and animals, Pathogenesis of the human opportunistic pathogen, Plant perceptions of plant growth-promoting, Technical advance: Spatio-temporal analysis of mitotic activity with a labile cyclin-GUS fusion protein, Dynamic analyses of the expression of the, Quorum-sensing cross talk: Isolation and chemical characterization of cyclic dipeptides from, Mechanism of auxin perception by the TIR1 ubiquitin ligase, Plant development is regulated by a family of auxin receptor F box proteins, ARF7 and ARF19 regulate lateral root formation via direct activation of LBD/ASL genes in Arabidopsis, Dual pathways for regulation of root branching by nitrate, Proceedings of the National Academy of Sciences, Earth, Atmospheric, and Planetary Sciences, www.pnas.org/lookup/suppl/doi:10.1073/pnas.1006740108/-/DCSupplemental, Opinion: To stop the next pandemic, we need to unravel the origins of COVID-19, Inner Workings: The quest to understand neutron stars and matter at the extremes, Journal Club: Smaller salamander species associated with smaller genomes. Again, Epo/E9 proved to be the most responsive element combination and provided the most useful profile in terms of low basal levels, high responsiveness and experimental reproducibility. Neural signatures of complex cognitive processing can occur in the absence of visual awareness, according to a case study of a person with distorted visual perception. Correspondence to T24 and MCF-7 tumor cells transiently transfected with the constructs in Figure 1 were incubated under 0.1% O2 for 24 h, followed by 7 h reoxygenation. The coordinating researcher (S. Di Saverio) invited six experienced surgeons (G. Augustin, A. Birindelli, B. To obtain
Combining gene therapy and radiotherapy protocols has the potential to overcome many of the limitations of adverse tumor biology on cancer treatment. Further improvements to the combinational promoter system may be achieved by optimizing the spacing between the elements, as well as their number and orientation and distance from the transcription start site. PubMed Google Scholar. Interestingly, besides being activated by IR, the E9 CArG enhancer also produced a 2.5-fold (±0.2) greater level of EGFP expression in hypoxic compared with normoxic T24 cells Figure 2. Prodrug activation was performed in air after hypoxic and/or IR treatment, and the efficacy of the Epo/E9 induction was compared with single component controls (Figure 5). First, three HRE units were inserted upstream of E9, then a further two HRE units making a total of five in the final enhancer Figure 1. Abrogation of apoptosis induced by DNA-damaging agents in human bladder- cancer cell lines with p21/WAF1/CIP1 and/or p53 gene alterations Int J Cancer 1996 68: 501–505, Ebert BL, Firth JD, Ratcliffe PJ .
Construction of the plasmid containing nine CArG elements (pE9-EGFP; Figure 1) used the method described in Scott et al.52 For the constructs containing both HREs and CArG elements, pE9-EGFP was digested with BglII, treated with calf intestinal alkaline phosphatase and the HREs were inserted directly upstream of the CArG elements. *, significantly different from E9; **, significantly different from the corresponding HRE (P < 0.05). Blockage of the vascular endothelial growth factor stress response increases the antitumor effects of ionizing radiation Cancer Res 1999 59: 3374–3378, Kuroki M et al.
The radiation-responsive promoter from the early growth response (Egr) 1 gene has been used successfully for selective transgene expression following radiation treatment.13 More recently, synthetic gene promoters based on the consensus sequence CC(A/T)6GG, known as CArG elements,14 have been shown to produce efficient and specific gene activation following IR in a gene therapy context.1516.
wrote the paper.
Control plates were supplied with the greatest concentration of ethanol used in the AHL treatments.
Novel chimeric gene promoters responsive to hypoxia and ionizing radiation.
The presence of hypoxic regions in solid malignancies is one of the most significant predictive factors affecting the response to radiotherapy in a range of human tumors (reviewed in Ref. Even if only a fraction (about 20%, results not shown) of the transient transfectants expressed the HRP, almost 90% cell death was measured. Conversely, after hypoxic and/or radiation treatment, selective toxicity was only seen when the stimuli-inducible promoters were employed.
The addition of a further five copies of the PGK1 HRE in the construct pPGK110/E9 did not significantly increase the hypoxia-inducibility in T24 cells (fold-induction 1.7 ± 0.3). Dual-specificity promoters have been described containing HREs in combination with estrogen responsive promoters,20 cytokine-inducible enhancers21 and hepatoma-associated enhancers.22 The results presented here indicate that combinations of HREs and CArG elements (also known as serum response elements or SREs) can act as gene enhancers, responding to hypoxic and radiation stimuli, at a level at least equivalent to that seen for the separate components Figures 2Figures 3Figures 4. These elements are found at varying distances from, and orientations to, the coding region of several hypoxia-regulated genes (reviewed in Ref. Mitochondrial reactive oxygen species trigger hypoxia-induced transcription Proc Natl Acad Sci USA 1998 95: 11715–11720, Chandel NS et al. Copyright © 2020 National Academy of Sciences. These data indicate that the use of such chimeric promoters may effectively regulate therapeutic gene expression within the tumor microenvironment in gene therapy strategies aimed at addressing the problem of hypoxia in radiotherapy. When Amanda Strike’s children became teenagers, she decided life needed a shakeup.
These findings, combined with the data presented here, indicate that the basal CMV promoter may be a good candidate for use in hypoxia- and radiation-targeted gene therapy constructs. Stabilization of wild-type p53 by hypoxia-inducible factor 1alpha Nature 1998 392: 405–408, Firth JD, Ebert BL, Pugh CW, Ratcliffe PJ . We do not capture any email address.
When T24 cells were transfected with the pCD4 plasmid, little or no toxicity was detected under any of the treatment conditions. Oxidative stress produced by suprahepatic occlusion and reperfusion Hepatology 1993 18: 881–889, Chandel NS et al. After hypoxic treatment, cells were left to reoxygenate for 7 h in air in a 37°C CO2 incubator before gene expression analysis. This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. Can gene therapy overcome the problem of hypoxia in radiotherapy? The present work has also highlighted some important considerations concerning the choice of promoter framework and marker gene. AND Oxic and anoxic enhancement of radiation-mediated toxicity by horseradish peroxidase/indole-3-acetic acid gene therapy Int J Radiat Biol 2002 78: 178–181, Article
The Epo/E9 combination was clearly the most robust, both in terms of absolute induction values and consistency of response in individual experiments. The Epo/E9 enhancer construct, shown to be particularly effective in earlier EGFP assays, was adopted for use in GDEPT. Hypoxia- and radiation-responsive enhancer components were placed immediately upstream of the basal human cytomegalovirus (CMV) gene promoter in all vector constructs (Figure 1).
A radiation-controlled molecular switch for use in gene therapy of cancer Gene Therapy 2000 7: 1121–1125, Nettelbeck DM, Jerome V, Muller R . The seedlings were washed with fresh 0.2× MS medium and for 12–14 h histochemically stained for GUS activity.
In previous studies, positive, negative and negligible responses have been observed under hypoxia in a number of different cell lines using this promoter.354445. In contrast, the full-length CMV promoter was not affected by the treatments in MCF-7 cells, while slightly down-regulated in T24 cells Figures 2Figures 3Figures 4Figures 5. Restriction sites in parentheses were deliberately deleted during the cloning procedure (BglII ‘locks’).
IAA-Asp content is significantly higher in the badc1 badc2 double mutant than in the wild type (WT). Microorganisms and their hosts communicate with each other through an array of signals. ISSN 1476-5462 (online), Novel chimeric gene promoters responsive to hypoxia and ionizing radiation, Friend or foe, the role of EGR-1 in cancer, Graphene oxide–hydroxyapatite nanocomposites effectively deliver HSV-TK suicide gene to inhibit human breast cancer growth, p53 activated by
All experiments were replicated at least three times.
Similarly, 2.1 (±0.3), 2.5 (±0.4) and 2.6 (±0.6)-fold inductions were seen in MCF-7 cells. Relative EGFP fluorescence was measured by fluorescence activated cell-sorting (FACS) analysis on a Becton Dickinson FACScan (excitation maximum: 488 nm, emission maximum: 507 nm). The unique physiology of solid tumors: opportunities (and problems) for cancer therapy Cancer Res 1998 58: 1408–1416, CAS
Inflammatory diseases ... IAA is associated with severe atherosclerosis and is thus most common in the abdominal aorta.
This observation has been independently confirmed in a range of other cell lines (N Chadderton and S Robinson, personal communication).
Surviving fractions were normalized for plating efficiency of cells exposed to HBSS only.
contributed new reagents/analytic tools; R.O.-C., C.D.-P., M.M.-T., R.E.d.R., J.C.-G., and J.L.-B. Image credit: NASA’s Goddard Space Flight Center. Transkingdom signaling based on bacterial cyclodipeptides with auxin activity in plants. Three different HREs were tested, from the human erythropoietin (Epo) gene, the murine phosphoglycerate kinase 1 (PGK1) gene, and a novel sequence based on the human vascular endothelial growth factor HRE (nVEGF). To give an indication of absolute EGFP expression values, net results from HRE and CArG constructs following sham, hypoxia or radiation treatments are shown in Table 1.