A. Latorre

Cytoskeleton and paclitaxel sensitivity in breast cancer : the role of β-tubulins

The antineoplastic effect of paclitaxel is mainly related to its ability to bind the β subunit of tubulin, thus preventing tubulin chain depolarization and inducing apoptosis. The relevance of the Class I β‐tubulin characteristics have also been confirmed in the clinical setting where mutations of paclitaxel‐binding site of β‐tubulin Class I have been related to paclitaxel resistance in non small cell lung and ovarian cancers. In the present study, we verified the hypothesis of a relationship between molecular alterations of β‐tubulin Class I and paclitaxel sensitivity in a panel of breast cell lines with different drug IC50. The Class I β‐tubulin gene cDNA has been sequenced detecting heterozygous missense mutations (exon 1 and 4) only in MCF‐7 and SK‐BR‐3 lines. Furthermore, the expression (at both mRNA and protein level) of the different isotypes have been analyzed demonstrating an association between low cell sensitivity to paclitaxel and Class III β‐tubulin expression increasing. Antisense oligonucleotide (ODN) experiments confirmed that the inhibition of Class III β‐tubulin could at least partially increase paclitaxel‐chemosensitivity. The hypothesis of a relationship between β‐tubulin tumor expression and paclitaxel clinical response has been finally verified in a series of 92 advanced breast cancer patients treated with a first line paclitaxel‐based chemotherapy. Thirty‐five percent (95% CI: 45–31) of patients with high Class III β‐tubulin expression showed a disease progression vs. only 7% of patients with low expression (35% vs. 7%, p < 0.002). Our study suggests that Class III β‐tubulin tumor expression could be considered a predictive biomarker of paclitaxel‐clinical resistance for breast cancer patients.

Paclitaxel, cisplatin and lonidamine in advanced ovarian cancer. A phase II study.

A potential way to improve the results obtained with the standard carboplatin/cisplatin (CDDP)-paclitaxel treatment regimen in advanced ovarian cancer is to incorporate a modulating agent such as lonidamine (LND). In fact, LND has been shown to revert the resistance to cisplatin and to potentiate cisplatin activity experimental models and in clinical studies. 35 consecutive patients with advanced ovarian cancer, not previously treated with chemotherapy were treated with paclitaxel at a dose of 135 mg/m(2) intravenously (i.v.) on day 1 (in a 3 h infusion) and cisplatin at a dose of 75 mg/m(2) iv on day 2 plus LND orally (p.o.) at a dose of 450 mg/die for 6 consecutive days starting two days before chemotherapy, every 3 weeks for six cycles. Complete plus partial responses were observed in 8 (80%) out of the 10 women with measurable disease. In the 25 patients with evaluable disease, only four clinical progressions were observed (16%). Median progression-free survival (PFS) and overall survival (OS) were 28.5 (95% confidence interval (CI) 22.2-34.8) and 46.5 (95% CI 32.4-60.00) months respectively. Grade 3-4 neutropenia was observed in 9 (26%) patients. Alopecia, nausea and vomiting (Grade 3) were observed in 33 (94%) and 5 (14%) patients, respectively. In conclusion, the combination of CDDP/paclitaxel plus LND is active and tolerable in the treatment of advanced ovarian cancer.

Subcutaneous recombinant interleukin-2 plus chemotherapy with cisplatin and dacarbazine in metastatic melanoma

The aim of our study was to verify the efficacy and tolerability of subcutaneous low doses of interleukin-2 with cisplatin and dacarbazine for malignant melanoma. 24 patients were included. The following schedule was used: cisplatin (CDDP) 100 mg/m2 day 1; darcarbazine (DTIC) 375 mg/m2 days 1-5; recombinant interleukin-2 (rIL-2) 4.5 million IU x 2/day days 13-17 and 20-24. The therapy was recycled every 28 days. 10 patients obtained clinical remission (42%), with 2 complete responses (8%) persisting for 12 and 15+ months, and 8 partial responses (35.5%) with a median duration of 5 months. Median survival of all 24 patients was 8 months, 13 months for responders and 6 months for non-responders. Responses were seen predominantly in lymph nodes (48%) and skin-soft tissue (38%), but were also seen in the liver (29%) and lung (14%). Treatment was relatively well tolerated and toxicity was mainly related to chemotherapy.

Revertant and potentiating activity of lonidamine in patients with ovarian cancer previously treated with platinum.

PURPOSE Lonidamine (LND) is an energolytic derivative of indazol-carboxylic acid that has been shown to enhance cisplatin (CDDP) activity in both sensitive (A2780) and resistant (A2780/Cp8) ovarian cancer cell lines. The aim of this study was to confirm the potentiating or reverting activity of LND on CDDP activity obtained in experimental models in a phase II study of advanced ovarian cancer patients previously treated with platinum-based regimens. PATIENTS AND METHODS Twenty-seven consecutive women with histologically proven and measurable ovarian cancer previously treated with platinum compounds were treated with CDDP plus LND. CDDP was administered at 1 mg/kg intravenously (IV) once weekly for 6 weeks and every 3 weeks thereafter until disease progression or toxicity. LND was administered at 450 mg daily (1 tablet every 8 hours) for the entire period of therapy starting 3 days before the first CDDP administration. In addition, a higher LND dosage was provided on the day of CDDP administration in an attempt to maximize the synergy of this drug with CDDP. RESULTS Ten patients achieved a complete response (CR) or partial response (PR) for an overall response rate of 37% (95% confidence interval [CI], 19% to 55%). In particular, responses were observed in five of 18 (28%) refractory or early relapsed patients (one CR and four PRs) and in five of nine patients (55%) in the late-relapsed group (two CRs and three PRs). Grade 3 or 4 anemia, leukopenia, and thrombocytopenia were observed in 19%, 15%, and 11% of patients, respectively, whereas seven of 27 patients (26%) showed LND-related myalgia. Grade 3 renal toxicity was observed in two patients (8%). Neurotoxicity, often concealed by LND-related myalgia, was recorded as grade 1 or 2 in six patients (22%) and as grade 3 in one (4%). CONCLUSION The 37% response rate observed in this study (28% in refractory or early-relapsed patients), suggests that the synergism between CDDP and LND observed in vitro against ovarian cancer cell lines can be clinically confirmed. However, larger series and randomized studies are needed to assess definitely the revertant activity of LND on CDDP-refractory patients.

Phase I/II study of gemcitabine plus mitoxantrone as salvage chemotherapy in metastatic breast cancer

The purpose of this study was to determine the maximum-tolerated dose of gemcitabine plus mitoxantrone in women with metastatic breast cancer (MBC) and to evaluate activity and toxicity of this combination in a phase II trial. Sixty-three patients with MBC, previously treated with chemotherapy including anthracycline and/or taxanes, were treated with mitoxantrone 10 or 12 mg m−2 intravenously on day 1 plus gemcitabine in escalating doses from 600 to 1200 mg m−2 intravenously on days 1 and 8, every 3 weeks. In phase I, on 23 patients entered on study, dose-limiting toxicity occurred at the dosage of 1200 mg m−2 gemcitabine and 10 mg m−2 mitoxantrone, with three out of five patients developing grade 4 neutropenia. In phase II, with gemcitabine administered at 1000 mg m−2 and mitoxantrone at 10 mg m−2, 12 (30%) out of 40 assessable patients responded, even if no complete response was obtained. Moreover, stable disease was observed in eight (20%) patients. The median time to treatment failure was 22 weeks (range, 2–33), and median survival was 42 weeks (range, 2–92). Grade 3 and 4 neutropenia were observed in 12 (30%) and one (2.5%) cases respectively; grade 3 thrombocytopenia was observed in two patients (5%), grade 2 mucositis in two patients (5%), grade 3 anaemia in two patients (5%), grade 3 alopecia in one patient (2.5%) and asymptomatic cardiotoxicity in three patients (8%), respectively. In conclusion, the doses of 10 mg m−2 (day 1) for mitoxantrone and 1000 mg m−2 for gemcitabine (days 1–8) every 3 weeks resulted active and safe in MBC. Further investigations in less heavily pretreated patients are warranted.

High efficacy of paclitaxel and doxorubicin as first-line therapy in advanced breast cancer: a phase I-II study.

Abstract The aim of this study was to define the maximum tolerated dose (MTD) of paclitaxel (TAX) in combination with doxorubicin (ADM). To evaluate the efficacy and tolerability of this combination, TAX was administered in escalating doses of 30 mg/m2, starting from 120 mg/m2, by 1 hour continuous infusion, per group of three patients; ADM was administered at a fixed dose of 50 mg/m2, 24 hours before administering TAX (phase I). The combination was recycled every 3 weeks. In phase II, TAX was administered at the MTD defined in phase I. Thirty-six women were enrolled. The MTD of TAX was 220 mg/m2. Objective responses were observed in 28/34 (82%) assessable patients. The median progression-free survival was 11.8 months and overall survival 27.8 months. The main clinical toxicity was neutropenia (grade III-IV) of short duration (94%). Two patients developed cardiac toxicity. The combination TAX+ADM is very effective in advanced breast cancer.

Subcutaneous rIL-2 in Advanced Melanoma and Kidney Carcinoma: Clinical and Biological Aspects

The aim of the present study was to verify the efficacy and tolerability of a treatment protocol using subcutaneous administration of rIL-2. Eighteen patients with metastatic disease were treated: 10 with renal carcinoma and 8 with malignant melanoma. The drug was administered as follows: 9 million UI/mq twice daily for two days followed by 1.8 million UI/mq twice daily for five days/week for six consecutive weeks. Of the 15 patients evaluated for clinical response, all completed treatment without rIL-2 dose modifications. No complete response was obtained. One patient with malignant melanoma and 1 with renal carcinoma (13%) had partial response (soft tissue and lymph nodes respectively) which lasted 3 and 4 months. Six patients with renal carcinoma and 3 melanoma patients had stable disease (60%) which lasted 6 months (median). Toxicity was moderate and spontaneously reversing after stopping treatment. Haemato-immunological modifications and pharmacokinetic study of this modality of rIL-2 treatment were also examined.

289PFISH TESTING OF HER2 IHC 1+ EARLY BREAST CANCER WITH UNFAVORABLE PROGNOSTIC FACTORS

ABSTRACT Aim: HER2-positive tumors are associated with a poor prognosis and a shortened disease-free and overall survival as well as with other unfavorable prognostic tumor characteristics (high histological grade, high proliferative index, negative or low estrogen receptor expression, etc.). HER2-positive tumors are also responsive to treatment with trastuzumab in reducing the risk of recurrence and improving survival. The aim of this study is to assess the incidence of HER2 gene amplification in selected tumors with adverse prognostic features which scored 1+ by immunohistochemistry (IHC). Methods: 75 women with infiltrating ductal carcinoma (IDC) and infiltrating lobular carcinoma (ILC) scoring 1+ by IHC were included. Forty-eight invasive breast carcinoma samples were selected according to unfavorable prognostic tumor characteristics and tested by FISH. HER2 amplification was evaluated using Vysis HER2/Cep17 probe (Path Vysion HER2 DNA Probe Kit®, Abbott Molecular, IL); ratio–based amplification was considered present when the HER2/Cep17 ratio was 2 or more and copy number-based amplification was considered present when the mean HER2 copy number was more than 6, in agreement with the ASCO/CAP/SIAPEC guidelines. Results: In 2013, 331 patients with invasive breast tumors were tested by IHC; 75 cases (23%) were scored 1+ of which 62 cases (19%) of IDC and 13 cases (4%) of ILC. Forty-eight invasive breast carcinoma samples (64%) were selected according to one or more unfavorable prognostic tumor characteristics; 22 out of 48 tumors (46%) showed high histological grade (G3); 27 cases (56%) had high proliferative index (Ki-67 ≥ 30%); 32 tumor samples (67%) were node-positive; and 29 cases (60%) showed vascular invasion. FISH was performed on 28 of the 1+ patients with adverse tumor characteristics and 6 IDC out of 48 (12.5%) showed HER2 amplification. Conclusions: Our preliminary retrospective data suggest that 6 patients out of 48 (12.5%) scoring 1+ by IHC show HER2 amplification, in agreement with the most recently published literature data. In order to not deny the benefit deriving from trastuzumab administration, in breast cancer patients showing IHC 1 + , it is advisable to test HER2 gene amplification by FISH. Disclosure: All authors have declared no conflicts of interest.

Rapid tumor shrinkage with lapatinib plus capecitabine in a patient with massive liver involvement

We present the case of a 58-year-old woman with breast cancer metastasizing to the liver after adjuvant chemotherapy. A liver biopsy confirmed metastatic lesions from breast cancer that were immunohistochemically positive for estrogen/progesterone receptors and HER2. After first-line treatment with trastuzumab and vinorelbine, the patient commenced therapy with capecitabine (1000 mg/m2 twice daily, days 1-14) and lapatinib (1250 mg/day). Three months after the administration of this combination therapy, the liver metastases had shrunk substantially. Lapatinib may have the potential to convert trastuzumab-refractory tumors to trastuzumab-sensitive tumors in HER2-positive breast cancer by upregulation of the cell surface expression of HER2. Further study will be needed to evaluate in the clinic the combination of lapatinib and an m-TOR inhibitor as a treatment approach in HER2 overexpressing breast cancer that shows a poor response to trastuzumab.

Lapatinib as salvage therapy in metastatic breast cancer: Preliminary results from an expanded access program

12006 Background: Lapatinib, a TKI of human epidermal growth factor receptor type 2 (HER2) and epidermal growth factor receptor (EGFR), is active in combination with capecitabine in women with HER2-positive metastatic breast cancer progressing after trastuzumab-based therapy. The primary end point of this single arm, open label trial, was to offer the approval expanded access to lapatinib, in order to provide potential clinical benefit. Secondary objectives were progression free survival, overall survival and evaluation of serious adverse events. Methods: Women affected by locally advanced or metastatic breast cancer with ErbB2 overexpressing tumours, measurable disease and baseline left ventricular ejection fraction ≥ 50% that had progressed after regimens including anthracyclines, taxanes and trastuzumab were included. Since march 2007, 18 patients were enrolled. They received lapatinib at a dosage of 1,250 mg daily continuously plus capecitabine at a dosage of 2,000 mg/m2 on days 1 through 14 of a 21-d...