Retargeting of adenoviral vector using basic fibroblast growth factor ligand for malignant glioma gene therapy

2005 ◽  
Vol 103 (6) ◽  
pp. 1058-1066 ◽  
Author(s):  
Weijun Wang ◽  
Nian-Ling Zhu ◽  
Jason Chua ◽  
Steve Swenson ◽  
Fritz K. Costa ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Cell Lines ◽  
Malignant Gliomas ◽  
Glioma Cells ◽  
Transduction Efficiency ◽  
Fibroblast Growth ◽  
Content Type ◽  
Fine Print

Object. Adenovirus vector (AdV)—mediated gene delivery has been recently demonstrated in clinical trials as a novel potential treatment for malignant gliomas. Combined coxsackievirus B and adenovirus receptor (CAR) has been shown to function as an attachment receptor for multiple adenovirus serotypes, whereas the vitronectin integrins (αvβ3 and αvβ5) are involved in AdV internalization. In resected glioma specimens, the authors demonstrated that malignant gliomas have varying levels of CAR, αvβ3, and αvβ5 expression. Methods. A correlation between CAR expression and the transduction efficiency of AdV carrying the green fluorescent protein in various human glioblastoma multiforme (GBM) cell lines and GBM primary cell lines was observed. To increase transgene activity in in vitro glioma cells with low or deficient levels of CAR, the authors used basic fibroblast growth factor (FGF2) as a targeting ligand to redirect adenoviral infection through its cognate receptor, FGF receptor 1 (FGFR1), which was expressed at high levels by all glioma cells. These findings were confirmed by in vivo study data demonstrating enhanced transduction efficiency of FGF2-retargeted AdV in CAR-negative intracranial gliomas compared with AdV alone, without evidence of increased angiogenesis. Conclusions. Altogether, the results demonstrated that AdV-mediated gene transfer using the FGF2/FGFR system is effective in gliomas with low or deficient levels of CAR and suggested that FGF2-retargeting of AdV may be a promising approach in glioma gene therapy.

Dependence of efficient adenoviral gene delivery in malignant glioma cells on the expression levels of the Coxsackievirus and adenovirus receptor

2000 ◽  
Vol 92 (6) ◽  
pp. 1002-1008 ◽  
Author(s):  
Katsuyuki Asaoka ◽  
Mitsuhiro Tada ◽  
Yutaka Sawamura ◽  
Jun Ikeda ◽  
Hiroshi Abe
Keyword(s):  
Gene Therapy ◽  
Cell Lines ◽  
Malignant Gliomas ◽  
Transduction Efficiency ◽  
Wild Type ◽  
Content Type ◽  
Expression Levels ◽  
Coxsackievirus And Adenovirus Receptor ◽  
Adenovirus Receptor ◽  
Fine Print

Object. Recombinant adenovirus is used as a competent vector in a wide spectrum of cancer gene therapies because of its high efficiency in gene delivery. To study the feasibility of gene therapy in malignant gliomas, the authors examined the antiproliferative effect of the adenovirally transduced wild-type p53 tumor suppressor gene by using 15 different high-grade glioma cell lines.Methods. Although growth suppression in association with a high adenoviral p53 transduction efficiency was seen in five of 15 cell lines, it was not observed in the remaining 10 cell lines. To clarify the underlying mechanism, we examined the expression levels of the Coxsackievirus and adenovirus receptor (CAR), which is the primary receptor for adenovirus, and of the integrins αvβ3 and αvβ5, which promote adenoviral internalization. The expression level of the CAR gene showed a close correlation to adenoviral gene transduction efficiency in the tested cell lines, whereas the expression levels of the integrins did not. The CAR expression was decreased by wild-type p53 transduction in U251MG cells harboring mutant p53 and increased by antisense inhibition of p53 in LN443 cells with endogenous wild-type p53.Conclusions. The results of this study indicate that CAR expression is a critical determinant of transduction efficiencies in adenovirus-based gene therapy for human malignant gliomas.

Targeted cytotoxic gene delivery to malignant gliomas via the fibroblast growth factor receptor

2000 ◽  
Vol 8 (4) ◽  
pp. 1-8
Author(s):  
Nian-Ling Zhu ◽  
Barbara A. Sosnowski ◽  
Berislav V. Zlokovic ◽  
Michael Ong ◽  
Thomas C. Chen
Keyword(s):  
Gene Therapy ◽  
Endothelial Cells ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Marker Gene ◽  
Malignant Gliomas ◽  
Glioma Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Fibroblast Growth ◽  
Fibroblast Growth Factor Receptors

Object The use of gene therapy for the treatment of malignant gliomas has been disappointing. In an effort to increase the viral transduction efficiency in delivering cytotoxic genes to malignant gliomas, the authors have used a novel retargeting schema that redirects the adenovirus to fibroblast growth factor receptors present on the cell surface of both proliferating glioma cells and glioma endothelial cells. Methods Using this targeted adenovirus, the authors demonstrated an increase in expression of the luciferase marker gene in human glioma cells and glioma endothelial cells. Transduction with the herpes simplex thymidine kinase gene resulted in greater cytotoxicity when treated with ganciclovir for both cell types. This increased cytotoxity was sustained for up to 6 days after administration of ganciclovir. Conclusions The mechanism of cytotoxicity was determined to be apoptosis by using the terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling assay. Increasing the specificity of adenovirus targeting may be crucial to decrease the number of adenoviral vectors necessary to create adequate levels of gene transfer in malignant gliomas, thus reducing the risk of vector-related immunogenicity and toxicity, as well as increasing the overall effectiveness of cytotoxic gene therapy.

Skull bone regeneration in primates in response to basic fibroblast growth factor

1999 ◽  
Vol 91 (5) ◽  
pp. 851-856 ◽  
Author(s):  
Yasuhiko Tabata ◽  
Keisuke Yamada ◽  
Liu Hong ◽  
Susumu Miyamoto ◽  
Nobuo Hashimoto ◽  
...  
Keyword(s):  
Growth Factor ◽  
Bone Regeneration ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Fibroblast Growth ◽  
Skull Defect ◽  
Skull Bone ◽  
Content Type ◽  
X Ray ◽  
Fine Print

Object. The feasibility of using a biodegradable hydrogel incorporating basic fibroblast growth factor (bFGF) to induce bone regeneration at the site of a skull defect in monkeys was investigated.Methods. Basic fibroblast growth factor was incorporated into a bioabsorbable hydrogel, which was prepared through glutaraldehyde crosslinking of gelatin. Following treatment of monkey skull defects measuring 6 mm in diameter (six defects/experimental group) with gelatin hydrogel incorporating bFGF, skull bone regeneration was evaluated using soft x-ray studies, dual x-ray absorptometry, and histological examinations.The water content of the hydrogels varied according to the glutaraldehyde concentration in the hydrogel preparation. Gelatin hydrogels incorporating 100 µg of bFGF significantly promoted bone regeneration and the skull defect was completely closed 21 weeks after implantation. This is in marked contrast with the effect of the same dose of bFGF in solution form. Bone mineral density (BMD) measured at the sites of skull defect was enhanced by the bFGF-incorporating hydrogels. The BMD enhancement was more prominent at lower water contents of hydrogel. Empty gelatin hydrogels neither induced nor interfered with skull bone regeneration.Conclusions. The findings of this study indicate that bFGF coupled with bioabsorbable hydrogel is a very promising tool to assist in the regrowth of bone at the site of a skull defect, which clinically has been recognized as almost impossible.

Negative autoregulation of fibroblast growth factor receptor 2 expression characterizing cranial development in cases of Apert (P253R mutation) and Pfeiffer (C278F mutation) syndromes and suggesting a basis for differences in their cranial phenotypes

2001 ◽  
Vol 95 (4) ◽  
pp. 660-673 ◽  
Author(s):  
Jonathan A. Britto ◽  
Rachel L. Moore ◽  
Robert D. Evans ◽  
Richard D. Hayward ◽  
Barry M. Jones
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Fibroblast Growth Factor Receptor ◽  
Growth Factor Receptor ◽  
Apert Syndrome ◽  
Fibroblast Growth ◽  
Content Type ◽  
Pfeiffer Syndrome ◽  
Negative Autoregulation ◽  
Fine Print

Object. Heterogeneous mutations in the fibroblast growth factor receptor 2 gene (FGFR2) cause a range of craniosynostosis syndromes. The specificity of the Apert syndrome—affected cranial phenotype reflects its narrow mutational range: 98% of cases of Apert syndrome result from an Ser252Trp or Pro253Arg mutation in the immunoglobulin-like (Ig)IIIa extracellular subdomain of FGFR2. In contrast, a broad range of mutations throughout the extracellular domain of FGFR2 causes the overlapping cranial phenotypes of Pfeiffer and Crouzon syndromes and related craniofacial dysostoses. Methods. In this paper the expression of FGFR1, the IgIIIa/c and IgIIIa/b isoforms of FGFR2, and FGFR3 is investigated in Apert syndrome (P253R mutation)— and Pfeiffer syndrome (C278F mutation)—affected fetal cranial tissue and is contrasted with healthy human control tissues. Both FGFR1 and FGFR3 are normally expressed in the differentiated osteoblasts of the periosteum and osteoid, in domains overlapped by that of FGFR2, which widely include preosseous cranial mesenchyme. Expression of FGFR2, however, is restricted to domains of advanced osseous differentiation in both Apert syndrome— and Pfeiffer syndrome—affected cranial skeletogenesis in the presence of fibroblast growth factor (FGF)2, but not in the presence of FGF4 or FGF7. Whereas expression of the FGFR2-IgIIIa/b (KGFR) isoform is restricted in normal human cranial osteogenesis, there is preliminary evidence that KGFR is ectopically expressed in Pfeiffer syndrome—affected cranial osteogenesis. Conclusions. Contraction of the FGFR2-IgIIIa/c (BEK) expression domain in cases of Apert syndrome— and Pfeiffer syndrome—affected fetal cranial ossification suggests that the mutant activation of this receptor, by ligand-dependent or ligand-independent means, results in negative autoregulation. This phenomenon, resulting from different mechanisms in the two syndromes, offers a model by which to explain differences in their cranial phenotypes.

scholarly journals Reduction of infarct volume and apoptosis by grafting of encapsulated basic fibroblast growth factor—secreting cells in a model of middle cerebral artery occlusion in rats

2003 ◽  
Vol 99 (6) ◽  
pp. 1053-1062 ◽  
Author(s):  
Kenjiro Fujiwara ◽  
Isao Date ◽  
Tetsuro Shingo ◽  
Hideyuki Yoshida ◽  
Kazuki Kobayashi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Middle Cerebral Artery ◽  
Fibroblast Growth Factor ◽  
Infarct Volume ◽  
Artery Occlusion ◽  
Fibroblast Growth ◽  
Content Type ◽  
And Control ◽  
Cerebral Artery Occlusion ◽  
Fine Print

Object. This study was conducted to evaluate the effects of grafting encapsulated basic fibroblast growth factor (bFGF)—secreting cells in rat brains subjected to ischemic injury. Methods. Two cell lines were used for encapsulated grafting in this experiment, namely, a bFGF-secreting cell line established by genetic manipulation of baby hamster kidney (BHK) cells, and a naive BHK cell line. Forty-seven Sprague—Dawley rats were used in this experiment. The animals were divided into the following three groups: those receiving grafts of encapsulated bFGF-secreting cells (BHK-bFGF group); those with grafts of encapsulated naive BHK cells (naive BHK group); and those with no grafts (control group). The authors implanted encapsulated cells into the right striatum of host rats in the BHK-bFGF and naive BHK groups. Six days after grafting, the host and control animals underwent permanent right middle cerebral artery occlusion (MCAO) with an intraluminal suture procedure. The infarct volume was evaluated using 2,3,5-triphenyltetrazolium chloride staining and computerized image analysis 24 hours after MCAO. Fragmentations of DNA in the host brains were analyzed using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling 12 hours after MCAO. The authors found that the infarct volume in the BHK-bFGF group was reduced by approximately 30% compared with that in the naive BHK and control groups. In the ischemic penumbral area, the number of apoptotic cells in the BHK-bFGF group was significantly decreased compared with that in the other groups. Conclusions. The grafting of encapsulated BHK bFGF-secreting cells protected the brain from ischemic injury. Encapsulation and grafting of genetically engineered cells such as bFGF-secreting cells is thus thought to be a useful method for protection against cerebral ischemia.

Antiproliferative effect of trapidil, a platelet-derived growth factor antagonist, on a glioma cell line in vitro

1990 ◽  
Vol 73 (3) ◽  
pp. 436-440 ◽  
Author(s):  
Jun-ichi Kuratsu ◽  
Yukitaka Ushio
Keyword(s):  
Growth Factor ◽  
Cell Lines ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Inhibitory Effect ◽  
Platelet Derived Growth Factor ◽  
Content Type ◽  
Glioma Cell Lines ◽  
Fine Print

✓ Platelet-derived growth factor (PDGF) is produced by glioma cells. However, there is heterogeneity among glioma cell lines in the production of PDGF. It has been demonstrated that U251MG cells produce a PDGF-like molecule while U105MG cells do not. Trapidil, a specific antagonist of PDGF, competes for receptor binding with PDGF. Therefore, the inhibitory effect of trapidil on the proliferation of glioma cells was investigated in vitro using two glioma cell lines. At 100 µg/ml, trapidil significantly inhibited the proliferation of U251MG cells (which produce the PDGF-like molecule). At the same trapidil concentration, the proliferation of U105MG cells (which do not produce the PDGF-like molecule) was not inhibited. The inhibitory effect of trapidil was remarkable on Days 3 and 4 of culture. After 4 days of incubation, the proliferation of U251MG cells was 46% of the control preparation. Trapidil enhanced the antitumor effect of 3-((4-amino-2-methyl-5-pyrimidinyl)ethyl)-1-(2-chloroethyl)-1-nitro-sourea (ACNU) against U251MG cells. The enhancing effect was highest on Days 4 and 6 of culture. After 6 days of incubation in the presence of 100 µg/ml trapidil and 1 µg/ml ACNU, the proliferation of U251MG cells was 18% of the control preparation. These findings suggest that trapidil interrupts the autocrine loop at the PDGF and PDGF-receptor level and that combination therapy with trapidil and ACNU may be useful in the treatment of glioma.

Expression of the basic fibroblast growth factor gene in mild and more severe head injury in the rat

1998 ◽  
Vol 89 (2) ◽  
pp. 297-302 ◽  
Author(s):  
Shu-Yuan Yang ◽  
Jian-Zhong Cui
Keyword(s):  
Brain Injury ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Severe Injury ◽  
Fibroblast Growth ◽  
Content Type ◽  
Injury Group ◽  
Fine Print

Object. The goal of this study was to investigate the relationship between basic fibroblast growth factor (bFGF) gene expression and neuropathological changes in the hippocampus after varying degrees of brain injury. Methods. Mild and severe brain injury in rats was produced by using Marmarou's method. There were 25 animals in each brain injury group and 25 additional animals served as controls. Basic fibroblast growth factor gene expression was investigated by means of RNA hybridization, in situ hybridization, immunohistochemical analysis, and histological analysis using hematoxylin and eosin staining. A 3.7-kb bFGF messenger (m)RNA was detected in the rat hippocampus in both control and injured rats. In the mild injury group its expression was increased at 12 hours after injury and peaked on the 3rd day. Neuronal degeneration in the hippocampal CA2 and CA3 sectors was maximum on that day. In the severe injury group, the expression of the bFGF gene was the same as that in the mild injury group at corresponding times, but the number of surviving neurons in the CA2 and CA3 sectors was much lower than in the mild injury group. In situ hybridization showed that the main cells that expressed bFGF mRNA were pyramidal and granulocytic neurons in all three experimental groups. The number of neurons expressing bFGF mRNA in the severe injury group was less than that in the mild injury group, but the intensity of expression was greater. Immunohistochemical staining showed that the number of neurons expressing the bFGF protein was less in the severe injury group than in the mild injury group. Conclusions. It is concluded that after mild injury there is a close relationship between the expression of the bFGF gene and the degree of histological change in the hippocampus; this indicates that as one of the growth factors, bFGF may participate in the protection and repair processes of neurons following brain injury. In severe injury there is a reduced expression of bFGF. The reason for this appears to be that more of the cells that have the potential to express bFGF have died, reducing the ability to express the bFGF gene. Conversely, it is possible that there may be an intrinsic insufficiency of expression of the gene, compatible with the known vulnerability of the hippocampus to many pathological conditions. Consideration should be given to supplying exogenous bFGF to protect the brain, particularly the hippocampus, after injury.

Immobilization of basic fibroblast growth factor on a platinum microcoil to enhance tissue organization in intracranial aneurysms

2005 ◽  
Vol 102 (1) ◽  
pp. 109-115 ◽  
Author(s):  
Takashiro Ohyama ◽  
Takuji Nishide ◽  
Hiroo Iwata ◽  
Hideki Sato ◽  
Mitsuaki Toda ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Coil Embolization ◽  
Control Group ◽  
Fibroblast Growth ◽  
Content Type ◽  
Tissue Organization ◽  
Platinum Coil ◽  
Fine Print

Object. To enhance tissue organization in an aneurysm lumen, the authors prepared a platinum microcoil carrying basic fibroblast growth factor (bFGF) and analyzed its effectiveness in the treatment of aneurysms. Methods. Ultrathin multiorganic layers were assembled on a platinum coil through successive deposition of cationic polyethylenimine and anionic heparin, and then bFGF was immobilized through an affinity interaction with heparin. The bFGF was effectively immobilized on the surface of the platinum coil without deterioration of the coil's mechanical properties. Coil embolization of aneurysms constructed using a canine common carotid artery was performed via the endovascular approach. The aneurysms together with parent arteries were harvested 2 weeks after coil embolization. Platinum coils unmodified, coated with heparin, or immobilized with heparin and bFGF were examined. The percentage of occlusion at the aneurysm orifice in animals treated with bFGF-immobilized coils (92.99 ± 7.94%) was significantly greater than that in animals treated with heparin-coated coils (57.26 ± 10.76%) or unmodified coils (52.86 ± 8.54%). The histological score of the aneurysms treated with bFGF-immobilized coils was also significantly greater than the scores in the control group. Conclusions. These results indicated that bFGF-immobilized microcoils may be beneficial in the obliteration of aneurysms.

Fibroblast growth factor receptor 3 mutation in nonsyndromic coronal synostosis: clinical spectrum, prevalence, and surgical outcome

2000 ◽  
Vol 92 (4) ◽  
pp. 631-636 ◽  
Author(s):  
Dominique Renier ◽  
Vincent El Ghouzzi ◽  
Jacky Bonaventure ◽  
Martine Le Merrer ◽  
Elizabeth Lajeunie
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Dna Analysis ◽  
Fibroblast Growth Factor Receptor ◽  
Growth Factor Receptor ◽  
Fibroblast Growth ◽  
Recurrent Point ◽  
Content Type ◽  
Coronal Synostosis ◽  
Fine Print

Object. A recurrent point mutation in the fibroblast growth factor receptor 3 gene that converts proline 250 into arginine has been reported recently in cases of apparently nonsyndromic coronal craniosynostosis. The goal of the present study was to examine the phenotype of patients in whom this mutation was present, to determine the prevalence of the condition, and to assess the functional and the morphological outcome of the surgically treated patients.Methods. A DNA analysis was performed in 103 children suffering from apparently isolated coronal synostosis, 41 of whom had bilateral and 62 of whom had unilateral disease. There were 31 boys and 72 girls in the study group. Sixty cases were sporadic and 43 were familial; the 43 familial cases arose in 33 unrelated families. The mutation was found in seven (12%) of 60 sporadic cases and in 24 (73%) of the 33 families. The functional and morphological results were assessed in all surgically treated patients who had at least 1 year of follow up and who were at least 3 years of age at the time of assessment. A comparison was made between patients with the mutation and those without.Conclusions. The most typical presentation was seen in girls and consisted of a bicoronal synostosis resulting in a severe brachycephaly associated with mild hypertelorism and marked bulging of the temporal fossae, which resulted in a huge enlargement of the upper part of the face. The most frequently associated extracranial anomaly was brachydactyly, identified either clinically or radiologically. Based on the proportion of bilateral and unilateral coronal synostoses, the present data indicate that the mutation is associated with more severe cases and that girls with the mutation are more severely affected than boys. The functional and morphological results were worse in patients in whom the mutation was present as compared with those in whom it was not.

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