Activity and safety of RAD001 (everolimus) in patients affected by biliary tract cancer progressing after prior chemotherapy: a phase II ITMO study.

BACKGROUND: Biliary tract cancer (BTC) is a highly lethal disease for which the best available therapy remains undetermined. The mammalian target of rapamycin (mTOR) pathway is up-regulated in several cancers, including BTC, and preclinical evidence indicates that mTOR inhibition may be effective in the treatment of BTC. We sought to evaluate the activity and tolerability of the mTOR inhibitor RAD001-everolimus-in patients with BTC progressing after prior chemotherapy. PATIENTS AND METHODS: This was an open-label, single-arm, phase II study (EUDRACT 2008-007152-94) conducted in eight sites in Italy. Patients with locally advanced, metastatic or recurrent BTC progressing despite previous chemotherapy received a daily oral dose of everolimus 10 mg administered continuously in 28-day cycles. The two primary end points were disease control rate (DCR) and objective response rate (ORR). Secondary end points were progression-free survival (PFS), overall survival (OS) and time-to-progression (TTP). RESULTS: Thirty-nine patients were enrolled. The DCR was 44.7%, and the ORR was 5.1%. One patient showed a partial response at 2 months and one patient showed a complete response sustained up to 8 months. The median (95% confidence interval) PFS was 3.2 (1.8-4.0) months, and the median OS was 7.7 (5.5-13.2) months. The median TTP was 2.0 (1.7-3.7) months. Most common toxicities were asthenia (43.6%), thrombocytopenia (35.9%), pyrexia (30.8%) and erythema, mainly of mild-to-moderate severity. Two patients required dose reduction due to adverse events. CONCLUSION: Everolimus demonstrated a favourable toxicity profile and encouraging anti-tumour activity. Further trials are needed to establish the role of everolimus in the treatment of BTC. EUDRACT 2008-007152-94.

Mutation analysis of the EGFR gene and downstream signalling pathway in histologic samples of malignant pleural mesothelioma.

BACKGROUND: As epidermal growth factor receptor (EGFR) is involved in the pathogenesis of malignant pleural mesotheliomas (MPMs), the anti-EGFR drugs may be effective in treating MPM patients. Mutations of the EGFR gene or its downstream effectors may cause constitutive activation leading to cell proliferation, and the inhibition of apoptosis and metastases. Consequently, molecular profiling is essential for select patients with MPM who may respond to anti-EGFR therapies. METHODS: After manual macrodissection, genomic DNA was extracted from 77 histological samples of MPM: 59 epithelioid, 10 biphasic, and 8 sarcomatoid. Epidermal growth factor receptor gene mutations were sought by means of real-time polymerase chain reaction (PCR) and direct sequencing, KRAS gene mutations by mutant-enriched PCR, and PIK3CA and BRAF gene mutations by direct sequencing. RESULTS: Gene mutations were identified in nine cases (12%): five KRAS, three BRAF, and one PI3KCA mutation; no EGFR gene mutations were detected. There was no difference in disease-specific survival between the patients with or without gene mutations (P=0.552). CONCLUSIONS: Mutations in EGFR downstream pathways are not rare in MPM. Although none of those found in this study seemed to be prognostically significant, they may support a more specific selection of patients for future trials.

Side effects associated with ultrarapid cytochrome P450 2D6 genotype among women with early stage breast cancer treated with tamoxifen.

BACKGROUND: The side effects of tamoxifen, a drug widely used for the treatment and the prevention of recurrence in patients with estrogen receptor positive breast cancers (ER+), have been reported in clinical trials, but to date no information is available on their possible association with an increased enzymatic activity of CYP2D6 (ultra-metabolizers, UMs). The aim of this study was therefore to evaluate the association between the presence of multiple functional CYP2D6 alleles and the occurrence of side effects. METHODS: 61 women with ER+ breast cancer receiving tamoxifen monotherapy were investigated in order to assess the relationships between CYP2D6 UM phenotype and side effects. Genotyping of 16 CYP2D6 polymorphisms was performed using a new DNA microarray technology. RESULTS: A highly significant difference was detected (41.2% of difference, 95% CI 6 - 61%, Fisher's exact test, p = 0.030) between the numbers of Ultrarapid Metabolizer patients (UM; high activity) with two or more adverse drug reactions to tamoxifen (7/9; 77.8%), compared to the number of Extensive Metabolizers (EM; normal activity), Intermediate Metabolizers (IM; reduced activity), and Poor Metabolizers (PM; no activity) with at least two side effects (19/52, 36.5%). A similar difference was also observed comparing the two groups (UM vs EM-IM-PM) for the number of side effects (median and inter quartile range, IQR: AM/EM/IM 1, IQR 0-2 vs. ULTRA 2, IQR 2-4; Mann-Whitney p = 0.005). CONCLUSIONS: Our results suggest a new association between CYP2D6 gene duplication and side effects to tamoxifen, indicating a possible role of CYP2D6 in their occurrence.

p27Kip1 is inactivated in human colorectal cancer by cytoplasmic localization associated with activation of Akt/PKB.

p27Kip1 is a nuclear member of the Kip/Cip family of cyclin-dependent kinase inhibitors and is a negative cell cycle regulator that is thought to play a role in tumour suppression. Reduced levels of p27Kip1 are frequent in human cancers and these have been associated with poor prognosis. We have analysed p27Kip1 expression and intracellular localization in 70 human colorectal cancers by western blotting and immunohistochemistry and the results related to Akt expression and clinical pathological parameters. p27Kip1 protein expression, as evaluated by western blotting, was absent or reduced in about 63% of colorectal cancers compared with both peritumoral and normal tissue. Cytoplasmic p27Kip1 was detected, by immunohistochemical analysis, in 30% (21 of 70) of cases indicating that translocation of p27Kip1 protein into the cytoplasm may be responsible for p27Kip1 inactivation. The analysis of phosphorylated Akt by western blotting indicated that it was expressed in 38% (8 of 21) of tumours showing cytoplasmic p27Kip1. Patients whose cancer presented accumulation of cytoplasmic p27Kip1 showed poorer outcomes for cancer-related relapse and survival. These results suggest that cytoplasmic p27Kip1 localization, associated or not with Akt activation, may contribute to colorectal tumorigenesis and metastasis and it may be useful as a negative prognostic factor for the outcome of patients with colorectal cancer.

Phase II study of pemetrexed in combination with carboplatin in patients with malignant pleural mesothelioma (MPM).

BACKGROUND: The aim of this study was to evaluate the activity and toxicity of pemetrexed and carboplatin combination as first-line chemotherapy in malignant pleural mesothelioma (MPM). PATIENTS AND METHODS: Patients with measurable advanced MPM and a zero to two Eastern Cooperative Oncology Group (ECOG) performance status (PS) were enrolled. The schedule was pemetrexed 500 mg/m(2) in combination with carboplatin area under the curve 5, every 21 days. In all, 76 patients were treated. Median age was 65 years; median ECOG PS was zero. RESULTS: Grade 3 hematological toxicity according to World Health Organization criteria was seen in 36 (47.3%) patients; grade 4 hematological toxicity in 5 (6.5%) patients. There were 16 (21%) partial responses and 3 (4%) complete responses, for an overall response rate of 19 (25%) [95% confidence interval (CI) 15.3-34.7]. In all, 29 (39%) (95% CI 28-48) patients reported stable disease. The median survival was estimated at 14 months. CONCLUSION: This combination of carboplatin and pemetrexed is moderately active and the toxicity is acceptable.

Prospective study on prognostic significance of DNA ploidy and Ki-67 expression in colorectal cancer.

The aim of the study was to correlate tumoral DNA ploidy and Ki-67 expression with therapy response, Overall Survival (OS), Disease Specific Survival (DSS) and Disease Free Survival (DFS). Three samples of colorectal cancer were collected from each patient. One sample of normal tissue was our internal control. DNA ploidy was evaluated by FACSCalibur cytometer and Ki-67 by immunohistochemistry. We studied 67 patients and we found aneuploidy in 65,7 percent of carcinoma with a Ki-67 median expression of 55 percent. After surgery and chemotherapy in 35 percent of the patients with aneuploid carcinoma and high proliferative activity (Ki-67 greater than 55 percent) there were no evidence of disease versus 100 percent of patients with DNA diploidy and low proliferative activity (Ki-67 less than 55 percent). Tumoral aneuploidy significantly correlated with lower OS, DSS and DFS (18 percent vs 86 percent at 30 months). Univariated analysis demonstrated a significant correlation between aneuploidy and develop disease progression (p=0,033, odd ratio=5.7), while the cut-off of 55 percent for Ki-67 expression did not correlate with OS, DSS and DFS. Preliminary results (the study is still in progress) seemed to suggest that DNA ploidy has a prognostic and predictive significance in colorectal carcinoma.

Production and pro-apoptotic activity of soluble CD95 ligand in pancreatic carcinoma.

We report here that the progression of pancreatic carcinomas in tumor patients is associated with increased serum levels of both the soluble forms of CD95 ligand (CD95L/FasL) and its receptor, CD95 (Fas). Shedding of proteolytically processed soluble CD95L was also observed in pancreatic carcinoma cells in vitro, thus identifying one possible source of CD95L in patients' sera. Because the secreted forms of both CD95 and CD95L have been implicated previously in protection of cells from CD95-mediated cell death, we assessed the effect of soluble CD95L in supernatants of pancreatic carcinoma cells on viability of Jurkat T lymphocytes. We describe that (a) supernatants derived from cultured pancreatic carcinoma cells caused apoptosis of Jurkat cells; (b) soluble tumor-derived CD95L contributed significantly to this effect; and (c) in comparison to Jurkat cells, pancreatic carcinoma cells themselves revealed increased resistance to apoptosis induction by autocrine soluble CD95L. These results are consistent with the notion that in the microenvironment of pancreatic tumors, tumor-derived shed CD95L exerts paracrine pro-apoptotic effects. In addition, because it is released at high levels into the bloodstream, soluble CD95L may have systemic effects in tumor patients that reach beyond the microenvironment of the tumor site.

Circannual variation of efficacy outcomes in patients with newly diagnosed metastatic colorectal cancer and treated with first-line chemotherapy.

Seasonal variation of baseline diagnosis (or clinical suspect) of stage I-III colorectal cancer patients has been repeatedly reported as an independent variable influencing overall survival. However, data are conflicting and no information is available about such a rhythm in advanced stage patients. To test whether a circannual rhythm of efficacy outcomes can be detected in this setting, we collected data about response rate (RR), progression-free survival (PFS), and overall survival (OS) to first-line chemotherapy of 1610 newly diagnosed metastatic patients treated at four independent centers. Responses to first-line chemotherapy were available for 1495 patients. A strong circannual rhythm in RR was evident, with the higher proportion of responding patients in the subgroup diagnosed in January (acrophase). At the time of data cutoff, 1322 patients progressed and 986 died, with median PFS and OS of 11 and 25.6 months, respectively. A circannual rhythmicity of the proportion of patients progressing at 6 months and surviving at 1 year was demonstrated, with acrophases located both in winter (February and January, respectively), similar to what reported for RR. Several interpretations about the genesis of this cyclic variation could be claimed: the rhythm in sunlight exposure and, as a consequence, of vitamin D serum levels and folate degradation, the variability in toxic effect intensity of chemotherapy, and the rhythm in the biological behavior of tumor cells. This observation is worth of further investigation both in preclinical and in clinical settings in order to better elucidate the underlying mechanisms.

Hepatic arterial infusion chemotherapy for unresectable confined liver metastases: prediction of systemic toxicity with the application of a scintigraphic and pharmacokinetic approach.

PURPOSE: The incorrect positioning of the arterial Port-a-Cath or the presence of anatomic or functional hepatic arteriovenous shunting may explain the occurrence of systemic toxicity of hepatic arterial infusion of floxuridine in patients with liver metastases. The aim of our study was to predict the occurrence of systemic toxic effects from this treatment using a scintigraphic and pharmacokinetic approach. METHODS: A group of 26 patients were studied. Before treatment, Tc-99m-labelled macroaggregated albumin arterial perfusion scintigraphy was performed to verify the correct positioning of the catheter, to evaluate the percentage of pulmonary uptake of the tracer, reflecting intrahepatic arteriovenous anatomic shunting, and to qualitatively assess the perfusion pattern of the metastases with respect to the normal liver parenchyma (SPECT images). Hepatic arteriovenous functional shunting was assessed through the bioavailability of intraarterially administered D-sorbitol. Treatment was then started and systemic toxic effects were evaluated according to WHO recommendations. RESULTS: No correlation was found between anatomic shunting (

Limits of aortic stop flow infusion chemotherapy in the treatment of advanced cancer.

BACKGROUND: Advanced and relapsed tumors remain a challenging disease with a poor and dismal prognosis. Our choice for inoperable tumors consists in a percutaneous treatment strategy involving intra-arterial chemotherapy and hemofiltration, with previous blood stop-flow, which allows high doses of Cisplatin-cisplatinum, cis-diammine-dichloroplatinum (CDDP) and Mitomycin C (MMC) in the tumor-bearing area with minimal systemic toxicity. METHODS: We analyse the morbidity and mortality associated with stop-flow in 20 patients with unresectable and/or metastatic thoraco- abdominal tumors, non responders to prior systemic chemotherapy. RESULTS: In our experience, the rate of major side effects of the procedure was 31% with a mortality of 5%. The side effects were related to the radiological procedure and to the chemotherapic treatment. A 74-year-old patient died for acute kidney toxicity within 15 days after the procedure. The other transient toxicity symptoms recorded were: nausea, vomiting, increasing of creatinine levels, diplopia and appearance of necrotic ulcer associated to chemotherapic drugs. Concerning the complications related to the radiological technique, the main problem was the rupture of the balloon stop-flow catheter in four patients. CONCLUSIONS: Stop-flow is a new procedure that could develop in the future, thanks to the possibility of obtaining a higher dose intensity of chemotherapic drugs in districts or organs affected by advanced tumors, with less systemic side effects. Unfortunately, the uncertain results in terms of increasing survival and the default of effective devices are to be resolved for a wider application of the procedure.

Extrahepatic regional chemotherapy: use of technetium-99m labeled macroaggregated albumin.

The purpose of this study was to verify the applicability of nuclear techniques with technetium-99m labeled macroaggregated albumin (Tc-99m-MAA) in extrahepatic regional chemotherapy. Of 98 patients in whom arterial Port-a-caths were implanted by transcutaneous access, 13 were treated by regional extrahepatic chemotherapy (breast, one; pancreas, four; kidney, one; uterus, three; vagina, two; bladder, two). In all 13 patients, Tc-99m-MAA was slowly infused intraarterially. The examination showed the perfusion of the area with the neoplasm and excluded the presence of important misperfusions of Tc-99m-MAA to the nearest areas. To detect the presence of an arteriovenous shunt with systemic misperfusion, an anterior image of the thorax was obtained in all patients and an index of misperfusion was calculated. In 12 patients, the index was < 5%; in one patient it was about 40%. In conclusion, our preliminary experience concerns the monitoring of intraarterial infusion chemotherapy of extrahepatic districts. In all 13 patients, we evaluated the correct positioning of the intraarterial catheter and the distribution pattern of the arterial flow, with a semiquantitative indication of arteriovenous shunting. This method gave us an instrument of study that was inexpensive, harmless, and free of collateral complications.

Combined regional and systemic chemotherapy by a mini-invasive approach for the treatment of colorectal liver metastases.

From February 1996 to December 1998, 95 patients affected with colorectal liver metastases underwent the positioning of an intraarterial hepatic catheter by a transcutaneous subclavian access, under local anesthesia. All patients were evaluated for catheter implantation complications. Moreover, 61 patients of 95 treated at our center were retrospectively evaluated for results of chemotherapy performed with two different schedules of hepatic artery infusion (HAI) combined with systemic chemotherapy (SC). Eleven patients (group A) were treated with combined SC (5-fluorouracil continuous infusion) and HAI (floxuridine). A subsequent 50 patients underwent HAI (floxuridine, 4 cycles) followed, if a response or stable disease were observed, by combined SC and HAI (group B). Three cases of aneurysm of subclavian artery occurred, which were treated by the positioning of a radiologic arterial stent and the reimplantation of the catheter by a femoral access. Thrombosis of the hepatic artery was registered in four cases. We observed 10.5% occurrence of dislocation of the catheter, which was always moved again in the hepatic artery. In group A, with 45% clinical objective response rate and 10% stable disease rate, median survival time and median time to extrahepatic progression were 9 and 6 months, respectively. In group B, we observed 44% clinical objective responses and 26% stable disease after HAI. Patients without disease progression and therefore submitted to sequential SC and HAI had a median survival time of 21 months and a median time to extrahepatic progression of 16 months. The development of the mini-invasive technique of implantation of an arterial port can avoid laparotomy for HAI. Percutaneous implantation of an arterial port has a low rate of technical complications. HAI followed by combined systemic and regional chemotherapy has good results in terms of survival and time to extrahepatic progression.

[High serum levels of Transforming Growth Factor-beta1, Interleukin-10 and Vascular Endothelial Growth Factor in pancreatic adenocarcinoma patients]. / Elevati livelli di Transforming Growth Factor-beta1 di Interleuchina-10 e di Vascular Endothelial Growth Factor nei sieri di pazienti affetti da adenocarcinoma pancreatico.

BACKGROUND: Pancreatic adenocarcinomas are among the most aggressive types of cancer with an extremely poor diagnosis. Since this type of cancer is not well amenable to chemo- and radiotherapy or immunotherapy, surgical resection remains the only feasible treatment to date. Transforming Growth Factor (TGF)-beta and Interleukin (IL)-10 are potent immunomodulators that have been shown to suppress several aspects of the immune response. Vascular Endothelial Growth Factor (VEGF) is a powerful angiogenic factor, recently thought to be involved in neoangiogenesis and metastasis spreading. Therefore the three cytokines may contribute, by different pathways, to immune escape and growth of tumor. This study was conducted to determine the possible significance of TGF-beta1, IL-10 and VEGF as markers for monitoring the clinical course of pancreatic adenocarcinoma patients. METHODS: Cytokine serum levels were measured in 30 pancreatic cancer patients and in 30 age and sex-matched healthy subjects. RESULTS: In comparison to serum concentrations in controls, TGF-beta1, IL-10 and VEGF levels were significantly elevated in all patients. Where the patients were divided by groups on the basis of the clinical stage of the disease, no differences were observed in TGF-beta1 levels among the groups. On the contrary, IL-10 and VEGF were more represented in stage IV patients than in stage II and III patients. In addition, the 14 patients who underwent surgical resection had postoperative cytokine serum levels markedly lower than those observed at diagnosis. CONCLUSIONS: Overall, the results suggest the importance of these markers in predicting the biological activity of the disease and suggest new targets for future rational therapies.

[CEA in the follow-up of breast carcinoma]. / Il CEA nel follow-up del carcinoma mammario.

One of the dominant aspects of cancer research over the past 10-15 years has been the search for a specific, sensitive marker for the various cancer types and one with a high predictive value. The importance and frequency of breast cancer, the demanding follow-up required in mastectomised patients and the fact that mammary carcinoma responds quite well to therapy with a fairly good survival rate as long as metastases and recurrences are promptly identified and treated encouraged this investigation into the value of CEA as a tumour marker in the follow-up of breast cancer. The problem was to discover whether. CEA would prove prompter than traditional investigations in identifying the appearance of recurrences or metastases or the increase of existing metastases and whether it provided a reliable indicator of the course of the disease and its response to treatment. Analysis of the results obtained in a personal series provided a negative answer to the first question and a positive one to the second. It is therefore concluded that only a well-planned and comprehensive follow-up, making use of all radiological, instrumental, laboratory and clinical aids will prove of any real benefit to mastectomised patients.

[Treatment of locally advanced breast neoplasms]. / Il trattamento delle neoplasie mammarie localmente avanzate.

One particularly interesting problem in the treatment of breast carcinomas relates to the strategy to be adopted in locally advanced forms. In 10-25% of cases surgery in itself offers no guarantee of radicality and the prognosis is particularly poor. Once considered suitable only for radiation treatment, such cases now tend to be treated by mixed therapy adopting different modes and sequences. The present paper fully and carefully examines current concepts and treatment possibilities.

[Current approach in the treatment of Zollinger-Ellison syndrome]. / Attuali orientamenti nel trattamento della sindrome di Zollinger-Ellison.

On the basis of six surgically operated patients, current thinking with regard to the treatment of Zollinger-Ellison syndrome is outlined. Total gastrectomy which for many years was the most common surgical intervention, is now confined to just a very specially indicated few cases because the introduction of anti-H2 gives good control of gastric secretion. The operation should therefore be targeted at removing the gastrinoma, helped in this by modern methodologies for pinpointing the neoplasia. Long-term however include a certain number of failures.

[Two cases of melanoma metastasis. Description and review of the literature]. / Due casi di metastatizzazione di melanoma. Descrizione e revisione della letteratura.

The metastases of malignant melanoma can appear after many years to primitive diagnosis and can involve many organs. There are reported two cases of metastatic melanoma presenting in a parotid gland and in the small bowel. The rarity of the metastatic melanoma to parotid gland and the difficulty of diagnosing intestinal involvement are discussed together with a review of the literature. The surgical treatment was justified by therapeutic and diagnostic aim, without positive influence on survival.

[CA 15-3 in the diagnosis and monitoring of carcinoma of the breast and its correlations with CEA]. / Il CA 15-3 nella diagnosi e nel monitoraggio del carcinoma della mammella e sue correlazioni con il CEA.

The effectiveness of marker CA 15-3 is assessed for the purposes of early diagnosis of breast cancer and its monitoring and data are compared with that of CEA. The results obtained in a series of 71 breast tumours show that CA 15-3 presents high positivity for T2 (66.2%) and possesses greater diagnostic sensitivity and accuracy than CEA.

Transaxillary access to perform hepatic artery infusion (HAI) for secondary or primitive hepatic tumors.

There is a renewed interest in locoregional chemotherapy for hepatic tumors; trials in progress are experimenting with new therapeutic protocols with an approach combining different systems of infusion (HAI and systematic) or with the use of HAI as adjuvant or neoadjuvant of the surgical treatment or cryosurgical treatment of the hepatic metastases from colo-rectal cancer. However, HAI is practicable principally with the implantation of a catheter in the hepatic artery (port of Infusaid) by laparotomic access. This intervention limits wide-scale use of the infusion method, traditionally less toxic and more efficient in terms of results than systemic treatment. Limited experience of percutaneous access for HAI required more catheterisation with repeated puncturing of the artery and later necessity of surgery in cases of HAI with continuous spraying. Motivated by the first experience of certain authors from Chiba University, we have devised a system of catheterisation of the hepatic artery with transcutaneous access, with subcutaneous port that allows the use of HAI without recourse to the usual intervention. Access is made through the left axillary artery; the positioning of the catheter is in the hepatic artery with possible embolization of the collateral or abnormal hepatic artery that could hamper complete diffusion of the drug to the liver, or increase to toxicity of the method. The implantation is done in day-surgery. In the cases performed up to now there have been no complications regarding the method and the catheters function all perfectly thanks to the collaboration of ematologists to avoid possible thrombosis of the catheters.

[Cytostatic therapy of metastatic breast cancer]. / La terapia citostatica del carcinoma mammario metastatico.

As complete as possible a picture of the current state of cytostatic chemotherapy in the treatment of metastatic breast carcinoma is offered. Attention is therefore mainly concentrated on application modalities of this form of therapy in relation to the clinical stage of the disease and to the use of other treatment possibilities. Methods and results of cytostatic therapy are then considered. The present situation is reviewed and the pharmacological and toxic aspects of prolonged antiblastic treatment are assessed. After outlining a number of special problems relating to the treatment of CNS and pleural metastases, the personal series is presented and results, which do not differ from other reported series, reported. In conclusion, stress is laid on the importance and effectiveness of cytostatic therapy in the treatment of metastatic breast carcinoma, but extended, controlled studies are also recommended.