Danilo Galizia

PARP1 expression drives the synergistic antitumor activity of trabectedin and PARP1 inhibitors in sarcoma preclinical models

BackgroundEnhancing the antitumor activity of the DNA-damaging drugs is an attractive strategy to improve current treatment options. Trabectedin is an isoquinoline alkylating agent with a peculiar mechanism of action. It binds to minor groove of DNA inducing single- and double-strand-breaks. These kinds of damage lead to the activation of PARP1, a first-line enzyme in DNA-damage response pathways. We hypothesized that PARP1 targeting could perpetuate trabectedin-induced DNA damage in tumor cells leading finally to cell death.MethodsWe investigated trabectedin and PARP1 inhibitor synergism in several tumor histotypes both in vitro and in vivo (subcutaneous and orthotopic tumor xenografts in mice). We searched for key determinants of drug synergism by comparative genomic hybridization (aCGH) and gene expression profiling (GEP) and validated their functional role.ResultsTrabectedin activated PARP1 enzyme and the combination with PARP1 inhibitors potentiated DNA damage, cell cycle arrest at G2/M checkpoint and apoptosis, if compared to single agents. Olaparib was the most active PARP1 inhibitor to combine with trabectedin and we confirmed the antitumor and antimetastatic activity of trabectedin/olaparib combination in mice models. However, we observed different degree of trabectedin/olaparib synergism among different cell lines. Namely, in DMR leiomyosarcoma models the combination was significantly more active than single agents, while in SJSA-1 osteosarcoma models no further advantage was obtained if compared to trabectedin alone. aCGH and GEP revealed that key components of DNA-repair pathways were involved in trabectedin/olaparib synergism. In particular, PARP1 expression dictated the degree of the synergism. Indeed, trabectedin/olaparib synergism was increased after PARP1 overexpression and reduced after PARP1 silencing.ConclusionsPARP1 inhibition potentiated trabectedin activity in a PARP1-dependent manner and PARP1 expression in tumor cells might be a useful predictive biomarker that deserves clinical evaluation.

Genotyping tumour DNA in cerebrospinal fluid and plasma of a HER2-positive breast cancer patient with brain metastases

Background Central nervous system (CNS) involvement contributes to significant morbidity and mortality in patients with human epidermal growth factor receptor 2 (HER2)-positive metastatic breast cancer (mBC) and represents a major challenge for clinicians. Liquid biopsy of cerebrospinal fluid (CSF)-derived circulating tumour DNA (ctDNA) harbours clinically relevant genomic alterations in patients with CNS metastases and could be effective in tracking tumour evolution. Methods In a HER2-positive mBC patient with brain metastases, we applied droplet digital PCR (ddPCR) and next-generation whole exome sequencing (WES) analysis to measure ctDNA dynamic changes in CSF and plasma collected during treatment. Results Baseline CSF-derived ctDNA analysis revealed TP53 and PIK3CA mutations as well as ERBB2 and cMYC amplification. Post-treatment ctDNA analysis showed decreased markers level in plasma, consistent with extra-CNS disease control, while increased in the CSF, confirming poor treatment benefit in the CNS. Discussion Analysis of ctDNA in the CSF of HER2-positive mBC is feasible and could represent a useful companion for clinical management of brain metastases.

Prolonged disease stability with trabectedin in a heavily pretreated elderly patient with metastatic leiomyosarcoma of the thigh and renal failure: a case report and review of the literature.

Leiomyosarcoma represents about 24% of all soft tissue sarcomas and can originate from retroperitoneum, uterus, or extremities. Adequate local control may be achieved with surgery and radiotherapy. In the presence of unresectable metastases either doxorubicin- or gemcitabine-based chemotherapy is the standard of treatment. Nevertheless, prognosis remains poor regardless of the selected chemotherapy regimen, and new effective therapeutic agents for patients with advanced leiomyosarcoma are needed. Trabectedin, a promising new DNA-damaging agent with a mechanism of action that is different from that of traditional alkylating agents, is approved in Europe for the treatment of patients with advanced soft tissue sarcoma, after failure of anthracyclines and ifosfamide, or who are unsuited to receive these agents and in combination with pegylated liposomal doxorubicin (PLD) for the treatment of patients with relapsed platinum-sensitive ovarian cancer. We present a case of a 76-year-old patient with progressive metastatic lung lesions from a previously resected primary leiomyosarcoma of the thigh and moderate renal failure, who achieved 17 months of disease stability during third-line treatment with trabectedin. Trabectedin was not associated with any cumulative toxicity and was consistently well tolerated for a total of 22 treatment cycles. Current evidence on trabectedin is also presented.

Confirmed Activity and Tolerability of Weekly Paclitaxel in the Treatment of Advanced Angiosarcoma

Background. In several prospective and retrospective studies, weekly paclitaxel showed promising activity in patients with angiosarcoma. Patients and Methods. Our study was originally designed as a prospective, phase II multicenter trial for patients younger than 75, with ECOG performance status 0–2, affected by locally advanced or metastatic angiosarcoma. Patients received paclitaxel 80 mg/m2 intravenously, at days 1, 8, and 15 every 4 weeks, until disease progression or unacceptable toxicity. Primary endpoint was objective response. Results. Eight patients were enrolled but, due to very slow accrual, the trial was prematurely stopped and further 10 patients were retrospectively included in the analysis. Out of 17 evaluable patients, 6 patients obtained an objective response (5 partial, 1 complete), with an objective response rate of 35% (95% confidence interval 17%–59%). Of note, five responses were obtained in pretreated patients. In the paper, details of overall survival, progression-free survival, and tolerability are reported. Conclusions. In this small series of patients with locally advanced or metastatic angiosarcoma, weekly paclitaxel was confirmed to be well tolerated and active even in pretreated patients.

Self-evaluation of duration of adjuvant chemotherapy side effects in breast cancer patients: A prospective study

We recently reported that self‐evaluation of the incidence and severity of treatment‐related side effects (TSEs) using a National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) v4.0‐based questionnaire was feasible and more informative than doctor reports in patients undergoing standard adjuvant chemotherapy for operable breast cancer. Here, we compare self‐ and doctor‐evaluated day of onset and duration of TSEs in the same population.

Abstract P5-11-01: Self-evaluation of duration of adjuvant chemotherapy side effects in breast cancer patients: A prospective study

Background : Collection and analysis of chemotherapy-related side-effects (CSE) is critical in the management of cancer patients (pts) both in experimental trials and in the clinical practice. Usually, most of the available conventional systems like the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) capture CSE severity but not their duration. Recently we observed that self-evaluation of CSE incidence and severity by a CTCAE v4.0-based questionnaire was feasible and potentially more informative than doctor reports in pts undergoing standard adjuvant chemotherapy (ACT) for operable breast cancer (Montemurro et al., JAMA Oncol 2; 445-452, 2016). Our questionnaire had also a section for reporting, for each of the considered CSE, day of onset, duration and whether it was still present at the time of the visit. Here we report the analysis of patient self-evaluation of CSE duration. Methods : The study prospectively enrolled 604 pts receiving ACT for operable breast cancer between January 2011 and October 2013 at 11 sites in Italy. CTCAE v4.0 definitions of grade of severity for nausea, vomiting, constipation, anorexia, dysgeusia, diarrhea, fatigue, pain, paresthesia, and dyspnea were translated into Italian and rephrased. Questionnaires were administered after the first and third cycle of chemotherapy. At each time-point, information on CSE was extracted from the medical charts and compared to patient questionnaires. Results : Overall 1177 questionnaires were collected, 596 after cycle 1 and 581 after cycle 3 of ACT. A median of 82% of the fields was completely filled-in. 594 and 573 pts-questionnaires had a corresponding MD-questionnaire. Comparison of CSE duration after cycle 1 of chemotherapy as self-assessed by pts versus that reported by doctors is summarized in the table. For all CSE, patient reported longer duration than doctors did. Comparisons of reports after cycle 3 were similar. Conclusions : self-reporting of CSE duration is feasible in patients receiving ACT after breast cancer surgery by using a CTCAE-derived questionnaire. Because doctors tend to underestimate both incidence and duration of CSE, patient-reported outcomes should be incorporated into the clinical practice because of the potential to provide a better estimate of the total burden of CSE. Supporte by a grant of the Rete Oncologica Del Piemonte e della Valle d9Aosta. Citation Format: Galizia D, Martinello R, Cagnazzo C, Foresto M, Gallizioli S, Longo V, Berchialla P, Solinas G, Calori A, Volpone C, Parola G, Tealdi G, Ballari A, Montemurro F. Self-evaluation of duration of adjuvant chemotherapy side effects in breast cancer patients: A prospective study [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P5-11-01.

Successful treatment of gemcitabine-induced acute interstitial pneumonia with imatinib mesylate: a case report

BackgroundGemcitabine is currently the standard chemotherapy for the adjuvant treatment of pancreatic cancer. This chemotherapeutic agent is generally well-tolerated, myelosuppression and gastrointestinal toxicity being common side effects. Nevertheless, gemcitabine-induced pulmonary toxicity has been rarely reported. Despite its low incidence, the spectrum of pulmonary injury is wide, including potentially fatal conditions.We report a case of acute interstitial pneumonia related to gemcitabine, completely solved with Imatinib Mesylate (IM).Case presentationThe patient was a 69-year-old man, who developed a hypoxemic respiratory distress during adjuvant treatment with gemcitabine for stage IIA pancreatic cancer. The nonspecific diffuse alveolar involvement found on computed tomography (CT), together with the negative tests for infectious aetiology and the continuing severe respiratory failure despite a long course of broad-spectrum therapy, suggested gemcitabine-induced acute pneumonia as the most likely diagnosis.Thus, after the failure of steroids and all other conventional therapies, the patient was treated with imatinib mesylate on the basis of its activity in the management of graft-versus-host-induced lung fibrosis. A follow-up CT scan of chest one month later showed complete resolution of pneumonia.ConclusionDespite the low frequency of serious pulmonary toxicity, gemcitabine widespread use warns clinicians to consider this life-threatening toxicity. The favourable clinical outcome with IM treatment was remarkable, warranting additional study of IM in the treatment of lung fibrosis.

350 Poly (ADP-ribose) Polymerase-1 (PARP-1) Inhibitors Potentiate Trabectedin Activity in Preclinical Models of Bone and Soft Tissue Sarcomas

Background: Trabectedin (TR) is a DNA-binding alkaloid approved in Europe for the treatment of soft tissue sarcoma as second/third line therapy. TR binds to the minor groove of DNA and interferes with gene transcription and nucleotide excision repair, inducing DNA double strand breaks (DSBs), and S/G2 cell cycle arrest. There is a strong clinical interest to increase TR activity by combination with other anti-cancer drugs. PARP-1 inhibitors (PARP1-i) which disable DNA base-excision repair mechanism causing the accumulation of DSBs, look like a worth to explore TR therapeutic partners. We focused our in vitro studies on the effects of the combination of TR with the PARP1-i olaparib (OL) and veliparib (VEL). Methods:We explored DNA damage by comet assay and phospho-histone H2AX determination and PARP-1 activity by polyADPribosylation assay in a panel of sarcoma cell lines treated with TR, OL,VEL and combinations. We evaluated cell viability after 72h treatment with escalating doses of TR (0−2nM), PARP1-i (VEL:0−80mcM; OL:0−20mcM), and their constant combinations. Following colony formation, cell cycle, apoptosis and DNA damage response were checked. Results: We demonstrated that TR-induced DNA damage and PARP1 activation in sarcoma sensitive cell line. The addition of PARP1i completely blocked basal and TR-induced PARP-1 activation. DNA damage response and checkpoints (ATM, ATR, BRCA1, CHK1, CHK2, p53) are activated after treatments with single agents and combinations inducing cell cycle arrest in S/G2 phase. This block was release after 48h in single agents-treated cells but not in combinations-treated cells leading to apoptotic cell death. Colony growth assay and viability tests revealed a strong synergism of the two drugs (Combination Index leyomiosarcoma > osteosarcoma undifferentiated pleomorphic > fibrosarcoma the most resistant). Determination of molecular markers of sensitivity and in vivo antitumoral activity studies are ongoing. Conclusions: Our results validate the biological rationale to combine TR and PARP1-i in sarcomas and suggest exploring this pharmacological approach in the clinical setting.