V. Bruce Grossie

Polyamine metabolism in carcinoma of the oral cavity compared with adjacent and normal oral mucosa

In this study, polyamine biosynthesis required for cellular proliferation showed elevated levels in neoplastic cells. Putrescine, spermidine, and spermine, as well as the rate-limiting enzyme ornithine decarboxylase, were measured to evaluate differences in tissue concentration in squamous cell carcinoma of the oral cavity, and in the normal adjacent, buccal, and retromolar trigone tissues. Mean polyamine levels (nanomoles per gram of tissue +/- standard error of the mean) were significantly elevated in tumor tissue at 136 +/- 42 nmol/g for putrescine compared with 41 +/- 9 nmol/g in adjacent, 25 +/- 5 nmol/g in buccal, and 41 +/- 14 nmol/g in retromolar trigone tissues. Tumor spermidine was 415 +/- 41 nmol/g compared with 192 +/- 34 nmol/g in adjacent, 184 +/- 34 nmol/g in buccal, and 214 +/- 63 nmol/g in retromolar trigone tissues. Tumor spermine was 461 +/- 41 nmol/g compared with 236 +/- 30 nmol/g in adjacent, 233 +/- 35 nmol/g in buccal, and 269 +/- 59 nmol/g in retromolar trigone samples. Ornithine decarboxylase activity was highly variable in tumor tissues. High levels of polyamines appear to be specific for this malignancy, whereas ornithine decarboxylase activity is not. Measurement of polyamine content may be useful in evaluating epithelial changes in the oral cavity.

Effect of difluoromethylornithine on host and tumor polyamine metabolism during total parenteral nutrition

Clinical and experimental data suggest that erythrocyte (RBC) polyamine (PA) levels are markers of tumor proliferation during total parenteral nutrition (TPN). The purpose of this experiment was to determine whether the inhibition of PA synthesis during TPN was greater in tumors than in normal host tissue. Rats bearing a subcutaneous fibrosarcoma were randomized to receive a chow diet (n = 5), TPN (n = 5), or TPN + difluoromethylornithine (DFMO) (an irreversible inhibitor of ornithine decarboxylase (ODC), at 1000 mg/kg body wt/day n = 4) for 6 days by continuous central venous infusion. TPN + DFMO resulted in a higher plasma albumin level and lower tumor ODC activity compared with chow feeding or TPN. Liver ODC activity was similar for the chow fed, TPN, and TPN + DFMO groups. RBC putrescine, tumor putrescine, and tumor spermidine levels were significantly lower in the TPN + DFMO group compared with the chow fed and TPN groups. RBC spermidine, RBC spermine, and tumor spermine levels were significantly increased with TPN + DFMO compared with TPN alone. DFMO did not produce diarrhea or weight loss. Increased RBC spermidine may indicate a toxic effect of DFMO on the tumor, resulting in leakage of tumor spermidine into the extracellular space. The data suggest that DFMO during TPN can selectively inhibit tumor PA synthesis and may improve host utilization of nutrients.

Evaluation of continuous-infusion alpha-difluoromethylornithine therapy for colorectal carcinoma

SummaryA total of 32 evaluable patients with measurable advanced colorectal carcinoma were treated with continuous-infusion alpha-difluoromethylomithine (DFMO) at a median daily dose of 8 g/m2 (range, 6–14 g/m2). DFMO was infused over 24 h daily for 28 days, followed by a rest period of 7 days. Of the 32 patients, 14 had received no prior chemotherapy. A total of 65 courses was given, with the median being 2 (range, 1–9 courses). None of the patients achieved a partial or complete response; however, 3 patients achieved a minor response and 14 had stable disease. The frequent toxic effects of DFMO included thrombocytopenia (which was dose-limiting), malaise, nausea, vomiting, reversible hearing loss, and diarrhea. Our data suggest that continuous-infusion DFMO therapy is feasible and results in only mild gastrointestinal toxicity. Although DFMO proved to be ineffective as a single agent in this trial, it could probably best be used in combination with cytotoxic agents known to enhance its antitumor activity in a preclinical setting.

Effects of Intralipid infusion on rat serum lipoproteins

The purpose of this investigation was to study the effects of continuous Intralipid® infusions on serum HDL and LDL levels in the rat. Male Fischer 344 rats were infused continuously via central venous catheter with 10% Intralipid® for 96 h and 5, or 2.5% Intralipid for 14 days. Blood samples were collected during the infusion period for total serum cholesterol, HDL-, and LDL-cholesterol measurements. Food intake was monitored during the studies. Total cholesterol, HDL-cholesterol and LDL-cholesterol levels were significantly elevated following 96 h of infusion with 10% Intralipid with food intake significantly decreased compared to a control group. In a second experiment, animals received a continuous infusion of either 5% Intralipid, 2.5% Intralipid or 0.45% saline for 14 days. Total cholesterol, HDL-cholesterol and LDL-cholesterol were significantly elevated following 14 days of infusion with 5% Intralipid group compared to controls but food intake remained constant for 12 days with no evident toxicity.

Differential Effects of Parenteral Nutrition on Tumor Growth and Erythrocyte Polyamine Levels in the Rat

The influence of total parenteral nutrition (TPN) on tumor growth and erythrocyte polyamine levels was evaluated in rats with a transplantable fibrosarcoma or a Ward colon tumor. During the experimental periods the fibrosarcoma grows exponentially when rats are fed chow ad libitum while the colon tumor reaches a plateau of its Gompertzian growth curve. A 12-day regimen of TPN resulted in an increased growth of the colon tumor but not the fibrosarcoma. The erythrocyte putrescine levels of fibrosarcoma-bearing rats and the levels of putrescine, spermidine, and spermine of Ward colon-tumor-bearing rats were significantly increased by TPN compared with similarly treated nontumor-bearing (NTB) rats. When the growth of the fibrosarcoma was slowed by feeding a restricted intake (RI) regimen, a subsequent 6-day regimen of TPN resulted in increased tumor growth. Erythrocyte polyamine levels of fibrosarcoma-bearing, RI-rats were elevated by TPN repletion. There was a consistent, significant, interaction between TPN and tumor presence on the erythrocyte putrescine levels in fibrosarcoma-bearing rats and the levels of all polyamines in Ward colon-tumor-bearing rats. The effects of TPN on tumor growth and erythrocyte polyamine levels of tumor-bearing rats may be dependent on the growth characteristics of the tumor. The data demonstrate that TPN consistently enhanced the tumor contribution to the erythrocyte putrescine pool.

Amelioration of thrombocytopenia with concomitant ornithine in sarcomabearing rats receiving high dose difluoromethylornithine

SummaryThe dose limiting toxicity of difluoromethylornithine (DFMO), when administered by continuous infusion, is thrombocytopenia. DFMO-induced antitumor activity and thrombocytopenia were time- and dosedependent up to 1700 mg/kg/d when administered continuously for 12 days. Concomitant ornithine administration (at selected molar ratios to DFMO) ameliorated thrombocytopenia induced by DFMO at a dose of 2000 mg/kg/day without adversely affecting its antitumor activity. The purpose of this study was to determine if ornithine could ameliorate the thrombocytopenia of higher DFMO doses and increase the efficacy of DFMO. Fischer 344 male rats with a transplantable sarcoma in the right flank were given 2000 and 3500 mg/kg/d DFMO alone or with ornithine at a molar ratio of 0.4 for 8 days by continuous infusion. Concomitant ornithine infusion overcame the thrombocytopenia that was induced by either dose of DFMO without reducing the antitumor activity against the sarcoma. The antitumor activity, tumor polyamine levels, and tumor S-adenosylmethionine decarboxylase activity did not consistently change with increasing doses of DFMO or with the addition of ornithine to the infusion regimen. These results demonstrate that the thrombocytopenia induced by doses of DFMO greater than 2000 mg/kg/d can be ameliorated without compromising the antitumor activity.

Effect of Ornithine in Parenteral Nutrition Regimens on Difluoromethylornithine-Induced Platelet Suppression and Changes in Tumor Polyamine Content

We have demonstrated that DFMO-induced thrombocytopenia can be ameliorated with concomitant ornithine (Orn) in chow-fed rats; a reversal in DFMO-associated tumor polyamine reduction and antitumor activity, however, was also evident. To determine the effect of Orn in total parenteral nutrition (TPN) regimens on DFMO-induced thrombocytopenia and changes in tumor polyamine concentrations, Ward-colon-tumor-bearing (WCT) rats were given TPN with arginine (ENA) or with ornithine substituted for arginine (ENO) alone or with DFMO (1.5 g/day) added directly to the infusate. After 4 days, the peripheral blood platelet counts for ENA (917 +/- 151 x 10(3)/mm3) or ENO (908 +/- 67 x 10(3)/mm3) were equivalent to those of chow fed rats (901 +/- 42 x 10(3)/mm3). ENA/DFMO rats had significant thrombocytopenia (607 +/- 185 x 10(3)/mm3), which was completely ameliorated for ENO/DFMO rats (939 +/- 111 x 10(3)/mm3). Peripheral white blood count, hematocrit, and other hematological parameters were not affected. Tumor putrescine content for ENA rats (46.9 +/- 8.7 nmol/g) was equal to that for chow-fed rats (44.8 +/- 6.2 nmol/g) and ENO rats (53.6 +/- 8.3 nmol/g). The reduction in tumor putrescine content for ENO/DFMO rats (19.6 +/- 6.9 nmol/g) was equivalent to that of ENA/DFMO rats (14.7 +/- 3.0 nmol/g). Tumor spermidine was reduced only for the ENA/DFMO rats while spermine was slightly elevated. Tumor spermine content for ENO/DFMO rats (57.2 +/- 12.0 nmol/g) was equal to that for ENO rats (65.6 +/- 8.7 nmol/g) but was significantly (p = 0.004) reduced when compared with rats receiving ENA/DFMO (89.4 +/- 20.4 nmol/g). The results of this study show that TPN with Orn substituted for arginine can be used with a chemotherapeutic dose of DFMO to ameliorate the thrombocytopenia. The DFMO-induced reduction in tumor putrescine content, however, was not affected when Orn was substituted for arginine in a parenteral nutrition regimen. These results suggest that the antitumor activity of DFMO would not be adversely affected by coadministering DFMO with a TPN regimen with Orn substituted for arginine.