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Adenovirus-based p53 gene therapy in ovarian cancer
312
Citations from the literature/International
Journal of Gynecology & Obstetrics 53 (19%) 305-316
was used for comparison. IC, was defined as the oligo concentration required for 50% growth reduction compared to untreated controls. Synergistic vs. antagonistic effects of oligo combinations were quantitated by combination indexes (CI) as calculated from median effect parameters by the methods of Chou and Talalay. Mean f SE ICso’s of c-myc and p53 antisenseoligos in CAOV3 and SKOVJ ranged from 1.0 i 0.2 to 9.7 f 1.3 CM. The ICse’s of c-myc oligos were consistently lower than corresponding p53 oligos in all cell lines (P < 0.034, t test). The tibroblast cell line was sensitive to antic-myc and combination anti-c-mytip oligos (It& = 1.5 f 0.6 and 1.4 * 0.2 CM, respectively), but not to anti-p53 oligos alone (ICss > 16 CM). Nonspecific toxicity was observed at concentrations of 16M for all cell lines except in BG-1, where maximal growth stimulation occurred al this concentration with anti-p53 oligos. Growth stimulation was also observed in BG-1 with antic-myc and anti+myc/p53 combinations at intermediate doses,with inhibition at higher doses. While c-myc/p53 combinations in CAOV-3 were synergistic (CI < 0.8), they were antagonistic in SKOV3 (CI > 3.2). Phosphorothioate oligos directed against c-myc and ~53 in different cell lines were shown to have both antiproliferative and stimulatory activity, as single agents and in combination, at concentrations that are achievable in vivo. Becauseof the complex patterns of effects, further in vitro studies are warranted before considering clinical trials with these agents in gynecologic cancers. p53 gem therapy in ovarian ameer Santoso J.T.; Tang D.-C.; Lane S.B.; Hung J.; Reed D.J.; Muller C.Y.; Carbone D.P.; Lucci J.A. III; Miller D.S.; Mathis J.M.
Adenovlnwbad
USA
GYNECOL ONCOL 199559/2 (171-178) Mutations of the ~53 tumor suppressor gene are the most common molecular genetic abnormality to be described in ovarian cancer. To determine the feasibility of mutant p53 as a mokcular target for gene therapy in ovarian cancer, we constructed an adenovirus vector containing the wild-type ~53 gene. The ability of this adenovirus construct (Ad-CMV-p53) to express ~53 protein was examined by Western blot analysis in the H358 lung cancercell line, which has a homoxygous deletion of the ~53 gene. The ability of the adenovirus vector system to infect ovarian cancer cells was tested using an adenovirus containing the &galactosidase reporter gene under the control of the CMV promoter (Ad-CMV-figal). The ovarian cancer cell line 2774,which contains an Arg273His ~53 mutation, was infected with Ad-CMV-&al, and the infected cells were assayedfor fi-galactosidaseactivity after 24 h. To test the ability of wild-type ~53 to inhibit cell growth, the 2774cell line was infected with Ad-CMV-p53 or Ad-CMV-fIgal, and the effect of theseagentson the growth of 2774cells was detennined using an in vitro growth inhibition assay.Western blot analysis of lysates from H358 cells infected with Ad-CMV-p53 showed expression of wild-type ~53 protein. When 2774 cells were infected with Ad-CMV-gal at a multiplicity of infection (m.o.i.) of 10 PFU/cell, >90?4 of cells showed f3-galactosidase activity, demonstrating that these cells are capable of efficient infection by the adenovirus vector. Growth of 2774 cells in-
fectedwith Ad-CMV-p53 was inhibited by >90% compared to noninfected celb.. The ability of the adenovirus vector to mediate high-level expression of infected genes and the inhibitory effect of Ad-CMV-p53 on the 2774 cell line suggests that the Ad-CMV-p53 could be further developed into a therapeutic agent for ovarian cancer. A novel pmgn&Ie factor In advanced Tamorv-tyeeniealcardmma Schlenger K.; Hockel M.; Mitxe M.; Schatfer U.; Weikel W.; Knapstein P.G.; Lambert A. DEU
GYNECOL ONCOL 1995 59/l (57-66) Objective: In the search for the optimal treatment of advancedcervical cancer, the identification of valid prognostic factors obtainable without histopathologic investigation of the entire tumor and the locoregional lymph nodesis of paramount interest. Tumor microvessel density has recently been demonstrated to correlate strongly with disease aggressivenessin breast cancer and other malignancies. Methods: We established a computerized image analysis system to quantify tumor microvascularity by using the closest-individual method, which determines the distribution of distances from random points within the tumor to the closest microvessel(DTCMV). Tumor microvascularity was assessedin paraffm sectionsof two cylindrical 2 x 20-mm core biopsies obtained transvaginally from the 12 and 6 o’clock positions of each tumor and then immunohistochemically stained for Factor VIII-related antigen. The oncologic relevance of tumor vasctdarity is studied in an open prospective trial. Results: Tumor vascularity was quantified in 42 patients with cervical cancers > 3 cm in largest diameter, FIG0 stages Ib-IVa. This new parameter representing pathophysiological tumor-host interactions was independent of various other patient and tumor characteristics, including age, FIG0 stage.,tumor sire, differentiation, lymph node metastasesand lymphatic spaceinvolvement. Thirty-nine patients were treated with curative intent either by primary surgery (n = 22) or radiation (n = 17).After a median observation time of 18 months (range 4-41 months), the patients with higher tumor vascularity (mean DTCMV < 83 m) had significantly shorter disease-free @ = 0.025) and overall @ = 0.032) survival probabilities than patients with lower tumor vascularity (mean DTCMV 2 83 pm). Cox regression analysis identified tumor vascularity as the strongest independent prognostic factor in this group of patients. Conclusions: The assessmentof tumor microvascularity by computerized image analysis of defined tumor biopsies could become a novel meansof predicting tumor aggressivenessin non-early cervical cancer. Conservative management of cImatkapeutIc-Induced
throol-
Imqtqm& in wmcn with gyuecoIogIccancers Fanning J.; Hilgers R.D.; Murray K.P.; Bolt K.; Aughenbaugb D.M. USA
GYNECOL ONCOL 199559/2 (191-193) During the course of four recent dose-intensechemotherapy trials, the routine practice of transfusing patients with platelet
Citations from the literature/International
Journal of Gynecology & Obstetrics 53 (19%) 305-316
was used for comparison. IC, was defined as the oligo concentration required for 50% growth reduction compared to untreated controls. Synergistic vs. antagonistic effects of oligo combinations were quantitated by combination indexes (CI) as calculated from median effect parameters by the methods of Chou and Talalay. Mean f SE ICso’s of c-myc and p53 antisenseoligos in CAOV3 and SKOVJ ranged from 1.0 i 0.2 to 9.7 f 1.3 CM. The ICse’s of c-myc oligos were consistently lower than corresponding p53 oligos in all cell lines (P < 0.034, t test). The tibroblast cell line was sensitive to antic-myc and combination anti-c-mytip oligos (It& = 1.5 f 0.6 and 1.4 * 0.2 CM, respectively), but not to anti-p53 oligos alone (ICss > 16 CM). Nonspecific toxicity was observed at concentrations of 16M for all cell lines except in BG-1, where maximal growth stimulation occurred al this concentration with anti-p53 oligos. Growth stimulation was also observed in BG-1 with antic-myc and anti+myc/p53 combinations at intermediate doses,with inhibition at higher doses. While c-myc/p53 combinations in CAOV-3 were synergistic (CI < 0.8), they were antagonistic in SKOV3 (CI > 3.2). Phosphorothioate oligos directed against c-myc and ~53 in different cell lines were shown to have both antiproliferative and stimulatory activity, as single agents and in combination, at concentrations that are achievable in vivo. Becauseof the complex patterns of effects, further in vitro studies are warranted before considering clinical trials with these agents in gynecologic cancers. p53 gem therapy in ovarian ameer Santoso J.T.; Tang D.-C.; Lane S.B.; Hung J.; Reed D.J.; Muller C.Y.; Carbone D.P.; Lucci J.A. III; Miller D.S.; Mathis J.M.
Adenovlnwbad
USA
GYNECOL ONCOL 199559/2 (171-178) Mutations of the ~53 tumor suppressor gene are the most common molecular genetic abnormality to be described in ovarian cancer. To determine the feasibility of mutant p53 as a mokcular target for gene therapy in ovarian cancer, we constructed an adenovirus vector containing the wild-type ~53 gene. The ability of this adenovirus construct (Ad-CMV-p53) to express ~53 protein was examined by Western blot analysis in the H358 lung cancercell line, which has a homoxygous deletion of the ~53 gene. The ability of the adenovirus vector system to infect ovarian cancer cells was tested using an adenovirus containing the &galactosidase reporter gene under the control of the CMV promoter (Ad-CMV-figal). The ovarian cancer cell line 2774,which contains an Arg273His ~53 mutation, was infected with Ad-CMV-&al, and the infected cells were assayedfor fi-galactosidaseactivity after 24 h. To test the ability of wild-type ~53 to inhibit cell growth, the 2774cell line was infected with Ad-CMV-p53 or Ad-CMV-fIgal, and the effect of theseagentson the growth of 2774cells was detennined using an in vitro growth inhibition assay.Western blot analysis of lysates from H358 cells infected with Ad-CMV-p53 showed expression of wild-type ~53 protein. When 2774 cells were infected with Ad-CMV-gal at a multiplicity of infection (m.o.i.) of 10 PFU/cell, >90?4 of cells showed f3-galactosidase activity, demonstrating that these cells are capable of efficient infection by the adenovirus vector. Growth of 2774 cells in-
fectedwith Ad-CMV-p53 was inhibited by >90% compared to noninfected celb.. The ability of the adenovirus vector to mediate high-level expression of infected genes and the inhibitory effect of Ad-CMV-p53 on the 2774 cell line suggests that the Ad-CMV-p53 could be further developed into a therapeutic agent for ovarian cancer. A novel pmgn&Ie factor In advanced Tamorv-tyeeniealcardmma Schlenger K.; Hockel M.; Mitxe M.; Schatfer U.; Weikel W.; Knapstein P.G.; Lambert A. DEU
GYNECOL ONCOL 1995 59/l (57-66) Objective: In the search for the optimal treatment of advancedcervical cancer, the identification of valid prognostic factors obtainable without histopathologic investigation of the entire tumor and the locoregional lymph nodesis of paramount interest. Tumor microvessel density has recently been demonstrated to correlate strongly with disease aggressivenessin breast cancer and other malignancies. Methods: We established a computerized image analysis system to quantify tumor microvascularity by using the closest-individual method, which determines the distribution of distances from random points within the tumor to the closest microvessel(DTCMV). Tumor microvascularity was assessedin paraffm sectionsof two cylindrical 2 x 20-mm core biopsies obtained transvaginally from the 12 and 6 o’clock positions of each tumor and then immunohistochemically stained for Factor VIII-related antigen. The oncologic relevance of tumor vasctdarity is studied in an open prospective trial. Results: Tumor vascularity was quantified in 42 patients with cervical cancers > 3 cm in largest diameter, FIG0 stages Ib-IVa. This new parameter representing pathophysiological tumor-host interactions was independent of various other patient and tumor characteristics, including age, FIG0 stage.,tumor sire, differentiation, lymph node metastasesand lymphatic spaceinvolvement. Thirty-nine patients were treated with curative intent either by primary surgery (n = 22) or radiation (n = 17).After a median observation time of 18 months (range 4-41 months), the patients with higher tumor vascularity (mean DTCMV < 83 m) had significantly shorter disease-free @ = 0.025) and overall @ = 0.032) survival probabilities than patients with lower tumor vascularity (mean DTCMV 2 83 pm). Cox regression analysis identified tumor vascularity as the strongest independent prognostic factor in this group of patients. Conclusions: The assessmentof tumor microvascularity by computerized image analysis of defined tumor biopsies could become a novel meansof predicting tumor aggressivenessin non-early cervical cancer. Conservative management of cImatkapeutIc-Induced
throol-
Imqtqm& in wmcn with gyuecoIogIccancers Fanning J.; Hilgers R.D.; Murray K.P.; Bolt K.; Aughenbaugb D.M. USA
GYNECOL ONCOL 199559/2 (191-193) During the course of four recent dose-intensechemotherapy trials, the routine practice of transfusing patients with platelet