Large-scale prospective genome-wide association study of oxaliplatin in stage II/III colon cancer and neuropathy.

BACKGROUND: The severity of oxaliplatin (L-OHP)-induced peripheral sensory neuropathy (PSN) exhibits substantial interpatient variability, and some patients suffer from long-term, persisting PSN. To identify single-nucleotide polymorphisms (SNPs) predicting L-OHP-induced PSN using a genome-wide association study (GWAS) approach. PATIENTS AND METHODS: A large prospective GWAS including 1379 patients with stage II/III colon cancer who received L-OHP-based adjuvant chemotherapy (mFOLFOX6/CAPOX) under the phase II (JOIN/JFMC41) or the phase III (ACHIVE/JFMC47) trial. Firstly, GWAS comparison of worst grade PSN (grade 0/1 versus 2/3) was carried out. Next, to minimize the impact of ambiguity in PSN grading, extreme PSN phenotypes were selected and analyzed by GWAS. SNPs that could predict time to recovery from PSN were also evaluated. In addition, SNPs associated with L-OHP-induced allergic reactions (AR) and time to disease recurrence were explored. RESULTS: No SNPs exceeded the genome-wide significance (P < 5.0 × 10-8) in either GWAS comparison of worst grade PSN, extreme PSN phenotypes, or time to recovery from PSN. An association study focusing on AR or time to disease recurrence also failed to reveal any significant SNPs. CONCLUSION: Our results highlight the challenges of utilizing SNPs for predicting susceptibility to L-OHP-induced PSN in daily clinical practice.

Large-scale prospective pharmacogenomics study of oxaliplatin-induced neuropathy in colon cancer patients enrolled in the JFMC41-1001-C2 (JOIN Trial).

BACKGROUND: Peripheral sensory neuropathy (PSN) is a dose-limiting toxicity of oxaliplatin-based chemotherapy. Several genetic markers have been shown to predict oxaliplatin-induced PSN; however, results remain to be validated in a large-scale and prospective pharmacogenomics study. PATIENTS AND METHODS: Among 882 patients enrolled in the JFMC41-1001-C2 (JOIN trial), which was designed to investigate the tolerability of adjuvant-modified FOLFOX6 (mFOLFOX6) in Japanese Patients with stage II or III colon cancers undergoing curative resection, 465 patients were eligible for this pharmacogenomics analysis. Twelve single-nucleotide polymorphisms (SNPs) were selected based on published data. The effect of each genotype on time to PSN onset was evaluated in all patients (n = 465) using the Cox proportional hazard model. For the association analysis between severity of PSN and 12 SNP markers, 84 patients who failed to complete 12 cycles of mFOLFOX6 from grade 0/1 PSN group were excluded because the termination of the protocol treatment had been caused by reasons other than PSN. RESULTS: Comparison of grade 0/1 PSN with grade 2/3 PSN or grade 3 PSN showed no significant associations with any of the 12 SNP markers after adjustment for total dose of oxaliplatin. Time-to-onset analysis also failed to reveal any significant differences. CONCLUSIONS: Our large-scale and prospective pharmacogenomics study of Japanese patients receiving protocol treatment of adjuvant mFOLFOX6 could not verify a role for any of the 12 SNP markers reported as being significantly associated with PSN. Considering the OR observed in this study (range: 0.76-1.89), further evaluation of these 12 SNP markers in the context of L-OHP-induced PSN is unlikely to be clinically informative.

A nested case-control study of stomach cancer in relation to green tea consumption in Japan.

To evaluate whether green tea consumption provides protection against stomach cancer, the relative risks (RRs) were calculated in the Japan Collaborative Study for Evaluation of Cancer Risk, sponsored by the Ministry of Health and Welfare (JACC Study). The study was based on 157 incident cases and 285 controls aged 40-79 years. Cox proportional hazards regression analysis was used to estimate the RRs for stomach cancer. It was found that green tea consumption had no protective effect against stomach cancer. After adjustment for age, smoking status, H. pylori infection, history of peptic ulcer, and family history of stomach cancer along with certain dietary elements, the risks associated with drinking one or two, three or four, five to nine, and 10 or more cups of green tea per day, relative to those of drinking less than one cup per day, were 1.3 (95% confidence interval (CI): 0.6-2.8), 1.0 (95% CI: 0.5-1.9), 0.8 (95% CI: 0.4-1.6), and 1.2 (95% CI: 0.6-2.5), respectively (P for trend=0.899). We found no inverse association between green tea consumption and the risk of stomach cancer.

A prospective study of stomach cancer death in relation to green tea consumption in Japan.

To evaluate whether green tea consumption provides protection against stomach cancer death, relative risks were calculated using Cox proportional hazards regression analysis in the Japan Collaborative Study for Evaluation of Cancer Risk, sponsored by the Ministry of Health and Welfare (JACC Study). The study was based on 30 370 men and 42 481 women aged 40-79. After adjustment for age, smoking status, history of peptic ulcer, family history of stomach cancer along with certain dietary items, the risks associated with drinking one or two, three or four, five to nine, and 10 or more cups of green tea per day, relative to those of drinking less than one cup per day, were 1.6 (95% CI: 0.9-2.9), 1.1 (95% CI: 0.6-1.9), 1.0 (95% CI: 0.5-2.0), and 1.0 (95% CI: 0.5-2.0), respectively, in men (P for trend=0.669), and 1.1 (95% CI: 0.5-2.5), 1.0 (95% CI: 0.5-2.5), 0.8 (95% CI: 0.4-1.6), and 0.8 (95% CI: 0.3-2.1), respectively, in women (P for trend=0.488). We found no inverse association between green tea consumption and the risk of stomach cancer death.

An immunosuppressive regimen using FTY720 combined with cyclosporin in canine kidney transplantation.

FTY720 induces apoptosis, specifically in lymphocytes, and prolongs allograft survival in rats and dogs. The purpose of this study was to define an effective range of FTY720 doses that could be combined with a suboptimal dose (10 mg/kg) of cyclosporin for canine kidney allograft recipients. The combination significantly prolonged allograft survival in all groups receiving FTY720 at a dose of 0.1, 0.3, 1.0, or 3.0 mg/kg. None of the recipients died due to notable side effects of the drug. In peripheral blood, the number of lymphocytes was extremely low, whereas the percentage of granulocytes increased during FTY720 administration. No significant difference in cyclosporin trough levels was observed between the cyclosporin-alone group and the combination groups. We conclude from the present study that FTY720 has a potent effect at an extremely low dose and a wide therapeutic window when combined with cyclosporin in canine kidney transplants.

Synergistic effect of nitric oxide and vasoactive intestinal peptide on bronchoprotection against histamine in anesthetized guinea pigs.

Vasoactive intestinal peptide (VIP) and nitric oxide (NO) are considered to be nonadrenergic, noncholinergic (NANC) inhibitory neurostransmitters in the airways. It seems likely that these neurotransmitters may be coreleased and act as functional antagonists against bronchoconstrictor stimuli. In the present study, we examined the synergistic effect of NO and VIP on bronchoprotection against histamine in anesthetized guinea pigs. The NO donor, S-nitroso-N-acetylpenicillamine (SNAP) significantly inhibited histamine-induced bronchoconstriction in a dose-dependent manner. VIP also inhibited histamine-induced bronchoconstriction in a dose-dependent manner, but this bronchoprotective effect was short-lived. Additionally, VIP (10(-9) M) had no significant bronchoprotective effect, but a subthreshold dose of SNAP (10(-7) M) significantly potentiated VIP (10(-9) M)-induced bronchoprotection against histamine. Moreover, SNAP (10(-7) M) significantly enhanced VIP (10(-7) M)-induced bronchoprotection for a longer period of time. On the other hand, VIP (10(-9) M) also significantly potentiated SNAP-induced bronchoprotection against histamine. In conclusion, combination therapy with NO donor and VIP receptor agonist may have important advantages in the treatment of bronchial asthma, and both NO and VIP may contribute in complementary fashion to the NANC-induced relaxant response in guinea pig airways.

Expression of carbohydrate-binding protein p33/41 in human tumor cell lines.

We previously reported a new type of lectin, p33/41 (annexin IV), which was isolated from a bovine tissue extract [Kojima, K. et al. (1992) J. Biol. Chem. 267, 20536-20539]. When the expression of p33/41 (annexin IV) was surveyed in the lysates of 39 human tumor cell lines by SDS-PAGE, followed by Western blot analysis with polyclonal anti-bovine p33/41 and monoclonal anti-annexin IV (Z016, Zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 cell lines were stained with Z016. These results together with those obtained with standard proteins, annexins IV and V, suggested that the monoclonal antibody, Z016, recognizes annexin V, but not p33/41 (annexin IV). Therefore, we performed cDNA cloning of human p33/41 (annexin IV) to prepare a recombinant protein and raised monoclonal antibodies against the protein. Northern blot analysis with the cDNA as a probe showed that a human colon cancer cell line, HT29, contains p33/41 (annexin IV) mRNA of two sizes, 2.0 and 3.0 kb. The two monoclonal antibodies, AS11 and AS17, against the recombinant protein generated were useful for flow cytometric analysis, ELISA, Western blot analysis and immunoprecipitation. Flow cytometric analysis with AS17 showed that p33/41 (annexin IV) is located in the cytoplasm of HT29 cells, but not on the cell surface. However, one of the cell surface proteins first labeled with biotin and then solubilized with a detergent was immunoprecipitated with AS17. The results suggest the existence of a membrane spanning form of p33/41 (annexin IV).

Inhibition of 5'-deoxy-5-fluorouridine phosphorolysis by acyclopyrimidinenucleosides in intestinal tissue homogenates.

This study examined the inhibitory effect of acyclopyrimidinenucleosides on 5'-deoxy-5-fluorouridine (5'-DFUR) phosphorolysis in intestinal tissue derived from rabbit, rat, mouse, and human. 5-Bromoacyclouridine, 5-fluoroacyclouridine, acyclouridine, and 5-nitroacyclouridine showed little or only moderate effect, but acyclothymidine [5-methyl-1-(2'-hydroxyethoxymethyl)uracil] showed strong inhibitory effect on 5'-DFUR phosphorolysis in intestinal tissue homogenates derived from human. In the absence of inhibitor (acyclothymidine), the Vmax of 5'-DFUR phosphorolysis was 2.66 mumol/min and the Km was 0.57 mM in human intestinal homogenates. The Vmax was unaltered by increased inhibitor concentration. The maximal inhibitory effect of acyclothymidine on 5'-DFUR phosphorolysis in rat homogenates was over 90%. The Ki/Km was 0.63 in human, 2.14 in rabbit, 1.09 x 10(-2) in rat, and 1.71 x 10(-2) in mouse. These data show that acyclothymidine is a competitive inhibitor of 5'-DFUR phosphorolysis, and that it can inhibit not only uridine phosphorylase but also thymidine phosphorylase.

Evaluation of a new lipophilic prodrug 3', 5'-dioctanoyl-5-bromodeoxyuridine (BrdU-C8) suspended in Lipiodol as a radiosensitizer for the treatment of AH136B tumor.

The effect of anti-cancer treatment on the AH136B tumor was studied using external beam irradiation in combination with a new oil-soluble agent, 3', 5'-dioctanoyl-5-bromodeoxyuridine (BrdU-C8), a lipophilic prodrug of BrdU. BrdU-C8 was dissolved in an oily lymphographic agent, Lipiodol (BrdU-C8/Lipiodol). BrdU-C8/Lipiodol is selectively accumulated in the neovasculature of the tumor and gradually releases BrdU. The AH136B tumor cell, a transplantable rat ascites hepatoma cell line, was implanted in the dorsal foot of rats. In vivo labeling index (L.I.) of the tumor cells after the intraarterial infusion of BrdU-C8/Lipiodol was significantly increased compared to the L.I. observed after intraarterial or intravenous infusion of BrdU. In addition, X-ray irradiation in combination with intraarterial infusion of BrdU-C8/Lipiodol significantly inhibited the tumor growth, indicating the increased radiosensitizing effect.

Anticancer effects of free polyunsaturated fatty acids in an oily lymphographic agent following intrahepatic arterial administration to a rabbit bearing VX-2 tumor.

The anti-hepatic cancer effects of three free polyunsaturated fatty acids (linoleic, alpha-linolenic, and gamma-linolenic acids) dissolved in an oily lymphographic agent, Lipiodol Ultra-Fluid (Lipiodol), following intrahepatic arterial administration were examined using a rabbit liver cancer model, VX-2. The tumor was inoculated into the subcapsular parenchyma of the liver of rabbits, and Lipiodol alone or Lipiodol containing each one of the free fatty acids was administered into the hepatic artery 14 days after inoculation. The rabbits were sacrificed 7 days after administration. Lipiodol containing one of the fatty acids selectively remained in the tumor area. Although VX-2 tumor grew extensively in both the untreated group and the group that received Lipiodol alone, growth of VX-2 tumor was greatly suppressed in the group that received Lipiodol containing the free fatty acid. Pathological observation also showed that Lipiodol containing the free fatty acid had an anticancer effect on VX-2 tumor growing in the liver of rabbits. Average survival days in the group treated with Lipiodol containing gamma-linolenic acid were significantly prolonged compared with those in the control groups. Although growth rates of the tumor at the death of rabbits were large in the control groups, VX-2 tumor shrank at death of five rabbits of six in the group treated with Lipiodol containing gamma-linolenic acid. These results suggest that the intrahepatic arterial administration of Lipiodol containing the free fatty acids is an effective method of delivery of these fatty acids as anticancer agents.

Intraarterial infusion of 5-fluoro-2-deoxyuridine-C8 dissolved in a lymphographic agent in malignant liver tumors. A preliminary report.

The drug 5-fluoro-2-deoxyuridine-C8 (FUdR-C8), one of the lipophilic prodrugs of FUdR, was dissolved in an oily lymphographic agent (Lipiodol Ultra Fluid, Andre Gelbe Laboratory, Paris, France; Ethiodol, Savage Laboratories, Melville, NY) and used for the intraarterial treatment of malignant liver tumors. From August 1985 to June 1988, 33 patients with hepatocellular carcinoma and 13 patients with metastatic liver tumors were treated with this agent at the Kumamoto University Hospital and its affiliated hospitals. The response rate (complete remission [CR] and partial remission [PR]) was 27.6% for hepatocellular carcinomas and 46.1% for metastatic liver tumors. The cumulative 1-year survival rate was 55.1% for hepatocellular carcinomas and 70.0% for metastatic liver tumors. More than a 50% decrease in the tumor marker level was observed in ten of 21 patients with hepatocellular carcinoma and in two of eight patients with metastatic liver tumors. The side effects, which were transient and controlled with conservative treatment, included fever, abdominal pain, nausea, vomiting, and acute gastritis.

Hydrolysis of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8) in rabbit and human plasma and its release from oily solution.

Hydrolysis of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8), a lipophilic prodrug of 5-fluoro-2'-deoxyuridine (FdUrd), 3'-octanoyl-5-fluoro-2'-deoxyuridine(3'-octanoyl FdUrd) and 5'-octanoyl-5-fluoro-2'-deoxyuridine (5'-octanoyl FdUrd) in the rabbit and human plasma and release of FdUrd-C8 and its hydrolyzed species from Lipiodol Ultra-Fluid (Lipiodol), an oily lymphographic agent, containing FdUrd-C8 were examined. The rates of hydrolysis of FdUrd-C8, 3'-octanoyl FdUrd and 5'-octanoyl FdUrd in the rabbit plasma were fast and almost the same among the three compounds; half lives were 2.4, 3.6 and 3.9 min for 5'-octanoyl FdUrd, FdUrd-C8 and 3'-octanoyl FdUrd, respectively. In the human plasma, however, the rates of hydrolysis were much different among the three compounds and were slower than those in the rabbit plasma; half lives were 11.5, 130 and 1020 min for 5'-octanoyl FdUrd, FdUrd-C8 and 3'-octanoyl FdUrd, respectively. Ratios of the rate constant of 5'-octanoyl FdUrd to that of 3'-octanoyl FdUrd were 1.6 and 85.7 in the rabbit plasma and in the human plasma, respectively. In a release study, although detected species in a release medium were different between the rabbit plasma and the human plasma and the amount of each species reflected the characteristics of esterase in each plasma, the total amounts of compounds released were almost the same both in the rabbit plasma and the human plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

Selective accumulation of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8) and sustained release of its active metabolites in VX-2 rabbit hepatoma following intraarterial administration of FdUrd-C8 solution in Lipiodol.

Selective accumulation/retention of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8) and sustained release of its active metabolites, 5-fluoro-2'-deoxyuridine (FdUrd) and 5-fluoro-2'-deoxyuridylate (FdUMP), in the rabbit hepatoma (VX-2) were achieved following intrahepatic arterial administration of FdUrd-C8 solution in Lipiodol. Though no significant difference in the FdUrd-C8 levels among the tumor and nontumorous liver was observed immediately after administration, slower elimination of FdUrd-C8 from the tumor (t 1/2 = 15.8 h) than that from nontumorous sites (t 1/2 = 3.8-4.2 h) resulted in selective retention of FdUrd-C8 (17- to 157-fold) in the tumor. Selectively higher levels of FdUrd and FdUMP in the tumor were also achieved (5- to 35-fold) and kept for 72 h after administration. The selective accumulation was also demonstrated in radioactivity distribution after administration of [6-3H]-FdUrd-C8. The ratio of radioactivity in the tumor divided by that in the blood (T/B ratio) was in a range of 870 to 5400 during a 15- to 1440-min period after administration. A trace of radioactivity was found in the stomach, duodenum, kidneys, and bone marrow. Roles of activation and deactivation enzymes on the selective distribution of FdUrd-C8 were also investigated. Esterase activity, which is responsible for the regeneration of FdUrd from FdUrd-C8, was relatively low in the tumor before administration and gradually increased after administration. Phosphorylase activity, which is related to phosphorolytic cleavage of FdUrd, in the tumor was about 3/5 as much as that in the nontumorous liver. These enzyme activities seem to play limited roles in the selective accumulation/retention and regeneration of the drug.

[Thermochemotherapy of unresectable pancreatic carcinoma (hot water and intra-arterial chemotherapy using noradrenaline)].

A 86-year-old man with unresectable pancreatic carcinoma was treated with thermochemotherapy. In most of the body and tail affected by the tumor, which did not show any cytotoxic damage after intraarterial chemotherapy, an enhanced antitumor effect was achieved by hyperthermia, which heated the tumor using hot water in a balloon placed on the pancreas, and which was delivered through a vinyl tube leading out of the body for 40 minutes while administration of antitumor agents mixed with noradrenaline was simultaneously given into the celiac artery. In this case, we observed that using noradrenaline with contrast material injected into the celiac artery it was possible to delineate the tumor area by computed tomography of the pancreas.

Selective anticancer effects of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine, a lipophilic prodrug of 5-fluoro-2'-deoxyuridine, dissolved in an oily lymphographic agent on hepatic cancer of rabbits bearing VX-2 tumor.

3',5'-Dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8), one of the lipophilic prodrugs of 5-fluoro-2'-deoxyuridine (FdUrd) was dissolved in an oily lymphographic agent (Lipiodol Ultra-Fluid), which had been studied as a carrier of the anticancer drug for hepatic cancer. The prodrug was administered into the left proper hepatic artery of rabbits bearing VX-2 tumor in the liver in order to examine the anticancer effects and possible adverse effects on nontumorous hepatic cells. Lipiodol or FdUrd-C8*Lipiodol selectively remained in the hepatic cancer area but disappeared from nontumorous parts of the liver 7 days after injection. Tumor growth rates in 1 week of the untreated group, a group given injections of 0.2 ml of Lipiodol alone, and groups given injections of 0.2 ml of Lipiodol containing 30, 50, 70, and 100 mg of FdUrd-C8 were 636, 436, 34.8, 14.9, -2.4, and -10.4% of the size at the time of treatment, respectively. Pathological observation also showed that FdUrd-C8 had a strong anticancer effect on VX-2 tumor growing in the liver of the rabbits. In contrast to the effect on the cancerous cells, that on nontumorous hepatic cells was very slight. In pathological observation, necrosis or degeneration of nontumorous hepatic cells was hardly observed. Plasma glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase levels temporarily rose 1 day after injection but returned to the initial levels within 7 days in all groups.

Peplomycin sulfate and pulmonary fibrosis: hydroxyproline, uronic acid, proline hydroxylase and glucosamine 6-phosphate synthetase in lungs of hamsters treated with peplomycin.

Effect of peplomycin sulfate (PLM) on pulmonary fibrosis was examined. Hydroxyproline, uronic acid, proline hydroxylase (EC 1.14.11.2) and glucosamine 6-phosphate synthetase (EC 2.6.1.16) in lungs of hamsters treated with PLM were studied and compared with those of hamsters treated with bleomycin (BLM). PLM, when administered intraperitoneally, one injection daily for 10 consecutive days, at either a high- (5 mg/kg) or low- (2.8 mg/kg) dosage-level, caused no significant increase of lung hydroxyproline and uronic acid as compared with controls. BLM on the other hand effected a significant increase in lung hydroxyproline on the high-dosage level (5 mg/kg) but not on the low-dosage level (2.8 mg/kg). In contrast, when administering PLM intratracheally, the concentrations of hydroxyproline in lungs increased 20% over the control levels. A transient increase of proline hydroxylase and glucosamine 6-phosphate synthetase also occurred shortly after the instillation. These increases were also observed in the corresponding groups treated with BLM, which confirmed the previous observations by other investigators. However, the magnitude of the increase was relatively lower in those values of PLM as compared with those of BLM. These data suggested that (1) PLM, when administered with multiple dosages intraperitoneally, showed no significant effect on the elevation of lung hydroxyproline; (2) PLM, when administered with a dose intratracheally, induced pulmonary fibrosis similar to that caused by BLM. However, the hydroxyproline accumulation in lungs of PLM-treated hamsters was less than in those of the BLM-treated; (3) The fibrotic effect on the lungs caused by either PLM or BLM was probably attributed to acceleration of the syntheses of collagen and acidic glycosaminoglycans.