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Dive into the research topics where Alessia Di Lorito is active.

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Featured researches published by Alessia Di Lorito.


Clinical Cancer Research | 2013

Effective Assessment of egfr Mutation Status in Bronchoalveolar Lavage and Pleural Fluids by Next-Generation Sequencing

Fiamma Buttitta; Lara Felicioni; Maela Del Grammastro; Giampaolo Filice; Alessia Di Lorito; Sara Malatesta; Patrizia Viola; Irene Centi; Tommaso D'Antuono; Roberta Zappacosta; Sandra Rosini; Franco Cuccurullo; Antonio Marchetti

Purpose: The therapeutic choice for patients with lung adenocarcinoma depends on the presence of EGF receptor (EGFR) mutations. In many cases, only cytologic samples are available for molecular diagnosis. Bronchoalveolar lavage (BAL) and pleural fluid, which represent a considerable proportion of cytologic specimens, cannot always be used for molecular testing because of low rate of tumor cells. Experimental Design: We tested the feasibility of EGFR mutation analysis on BAL and pleural fluid samples by next-generation sequencing (NGS), an innovative and extremely sensitive platform. The study was devised to extend the EGFR test to those patients who could not get it due to the paucity of biologic material. A series of 830 lung cytology specimens was used to select 48 samples (BAL and pleural fluid) from patients with EGFR mutations in resected tumors. These samples included 36 cases with 0.3% to 9% of neoplastic cells (series A) and 12 cases without evidence of tumor (series B). All samples were analyzed by Sanger sequencing and NGS on 454 Roche platform. A mean of 21,130 ± 2,370 sequences per sample were obtained by NGS. Results: In series A, EGFR mutations were detected in 16% of cases by Sanger sequencing and in 81% of cases by NGS. Seventy-seven percent of cases found to be negative by Sanger sequencing showed mutations by NGS. In series B, all samples were negative for EGFR mutation by Sanger sequencing whereas 42% of them were positive by NGS. Conclusions: The very sensitive EGFR-NGS assay may open up to the possibility of specific treatments for patients otherwise doomed to re-biopsies or nontargeted therapies. Clin Cancer Res; 19(3); 691–8. ©2012 AACR.


Journal of Thoracic Oncology | 2016

ALK Protein Analysis by IHC Staining after Recent Regulatory Changes: A Comparison of Two Widely Used Approaches, Revision of the Literature, and a New Testing Algorithm

Antonio Marchetti; Alessia Di Lorito; Maria Vittoria Pace; Manuela Iezzi; Lara Felicioni; Tommaso D’Antuono; Giampaolo Filice; L. Guetti; Felice Mucilli; Fiamma Buttitta

Introduction: Recent regulatory changes have allowed the diagnostic use of immunohistochemical (IHC) analysis for the identification of patients with non–small cell lung cancer who are eligible for treatment with anaplastic lymphoma receptor tyrosine kinase (ALK) inhibitors. The U.S. Food and Drug Administration has approved the VENTANA ALK (D5F3) CDx Assay (Ventana Medical Systems, Tucson, AZ) as companion diagnostics, and the Italian Medicines Agency has recognized IHC analysis as a diagnostic test indicating an algorithm for patient selection. Methods: On the basis of the new regulations, we compared two commonly used IHC assays on 1031 lung adenocarcinomas: the VENTANA ALK (D5F3) CDx Assay with the OptiView Amplification Kit (Ventana Medical Systems) and a standard IHC test with the clone 5A4 (Novocastra, Leica Biosystems, Newcastle Upon Tyne, United Kingdom) along with their interpretative algorithms. Fluorescence in situ hybridization (FISH) was performed in all cases. Next‐generation sequencing was performed in FISH/IHC analysis–discordant samples. Results: FISH gave positive results in 33 (3.2%) cases. When FISH was used as a reference, the VENTANA ALK (D5F3) CDx assay had a sensitivity of 90.9% ± 2.6%, a specificity of 99.8% ± 0.6%, and positive and negative predictive values of 93.8% ± 2.1% and 99.7% ± 0.6%, respectively. The clone 5A4–based IHC test showed a sensitivity of 90.9% ± 2.6%, a specificity of 98.3% ± 1.3%, and positive and negative predictive values of 63.8% ± 4.2% and 99.7% ± 0.6%, respectively. Five cases with IHC analysis/FISH‐discordant results in our series were analyzed together with those previously reported in the literature. Overall, data from 35 patients indicate a response rate to ALK inhibitors in 100% of FISH‐negative/IHC analysis–positive cases (seven of seven) and 46% of FISH‐positive/IHC analysis–negative cases (13 of 28), respectively. Conclusions: Our results confirm the difficulty in managing an IHC test without amplification in the absence of confirmatory FISH analysis, as well as the possibility of performing a direct diagnosis in approximately 90% of patients by the VENTANA ALK (D5F3) CDx Assay. On the basis of the recent regulatory changes, the data that have emerged from the literature, and the results of the present study, a new algorithm for ALK assessment in non–small cell lung cancer has been devised.


Lung Cancer | 2016

Validation of a new algorithm for a quick and easy RT-PCR-based ALK test in a large series of lung adenocarcinomas: Comparison with FISH, immunohistochemistry and next generation sequencing assays.

Antonio Marchetti; Maria Vittoria Pace; Alessia Di Lorito; Sara Canarecci; Lara Felicioni; Tommaso D’Antuono; Marcella Liberatore; Giampaolo Filice; L. Guetti; Felice Mucilli; Fiamma Buttitta

OBJECTIVES Anaplastic Lymphoma Kinase (ALK) gene rearrangements have been described in 3-5% of lung adenocarcinomas (ADC) and their identification is essential to select patients for treatment with ALK tyrosine kinase inhibitors. For several years, fluorescent in situ hybridization (FISH) has been considered as the only validated diagnostic assay. Currently, alternative methods are commercially available as diagnostic tests. MATERIAL AND METHODS A series of 217 ADC comprising 196 consecutive resected tumors and 21 ALK FISH-positive cases from an independent series of 702 ADC were investigated. All specimens were screened by IHC (ALK-D5F3-CDx-Ventana), FISH (Vysis ALK Break-Apart-Abbott) and RT-PCR (ALK RGQ RT-PCR-Qiagen). Results were compared and discordant cases subjected to Next Generation Sequencing. RESULTS Thirty-nine of 217 samples were positive by the ALK RGQ RT-PCR assay, using a threshold cycle (Ct) cut-off ≤35.9, as recommended. Of these positive samples, 14 were negative by IHC and 12 by FISH. ALK RGQ RT-PCR/FISH discordant cases were analyzed by the NGS assay with results concordant with FISH data. In order to obtain the maximum level of agreement between FISH and ALK RGQ RT-PCR data, we introduced a new scoring algorithm based on the ΔCt value. A ΔCt cut-off level ≤3.5 was used in a pilot series. Then the algorithm was tested on a completely independent validation series. By using the new scoring algorithm and FISH as reference standard, the sensitivity and the specificity of the ALK RGQ RT-PCR(ΔCt) assay were 100% and 100%, respectively. CONCLUSIONS Our results suggest that the ALK RGQ RT-PCR test could be useful in clinical practice as a complementary assay in multi-test diagnostic algorithms or even, if our data will be confirmed in independent studies, as a standalone or screening test for the selection of patients to be treated with ALK inhibitors.


Diagnostic Cytopathology | 2013

Molecular alterations in endometrial archived liquid‐based cytology

Alessia Di Lorito; Sandra Rosini; Eleonora Falò; Sarah Gustapane; Madalena Gomes; José Luis Costa; Fernando Schmitt

Endometrial cancer is one of the most common gynecological malignancy worldwide and its prevalence is increasing. The introduction of liquid‐based cytology (LBC) and endoflower dispositive in routine practice gives the possibility to examine endometrial cells by cytological diagnosis and may also release the opportunity to study molecular alterations, in endometrioid type cancer in which carcinogenesis is well known. We gathered 72 cases of endometrial LBC samples and corresponding formalin‐fixed paraffin‐embedded (FFPE) blocks, collected from 2004 to 2010. DNA was isolated from both samples using standard protocols. DNA quality and quantity were assessed using Nanodrop and BIOMED2 multiplex PCR. Mutations in exon 5 of PTEN and exon 20 of PI3K were studied using Sanger sequencing. DNA with good quality and amount was isolated from 67/72 FFPE cases. In these samples, two cases were found to harbor mutations in exon 5 of PTEN. No PI3K mutations were identified. LBC samples were then assessed to verify the concordance with the FFPE DNA results. The results obtained were concordant, that is the wild type cases in FFPE were also wild type in LBC and vice versa for the mutated case. Unfortunately, the second case of mutation in PTEN could not be confirmed in LBC due to low amount of DNA obtained. Detection of molecular alterations in LBC will open a new era for the detection in asymptomatic women of precursor lesions that could evolve into cancer and for endometrial cancer diagnosis and screening in selected high‐risk women. Diagn. Cytopathol. 2013.


Journal of Thoracic Oncology | 2017

Evaluation of NGS and RT-PCR methods for ALK rearrangement in European NSCLC patients: Results from the ETOP Lungscape Project

Igor Letovanec; Stephen Finn; Panagiota Zygoura; Paul Smyth; Alex Soltermann; Lukas Bubendorf; Ernst-Jan M. Speel; Antonio Marchetti; Daisuke Nonaka; Kim Monkhorst; Henrik Hager; Miguel Martorell; Aleksandra Sejda; Richard T. Cheney; Javier Hernández-Losa; Eric Verbeken; Walter Weder; Spasenija Savic; Alessia Di Lorito; Atilio Navarro; Enriqueta Felip; Arne Warth; Paul Baas; Peter Meldgaard; Fiona Blackhall; Anne-Marie C. Dingemans; Hendrik Dienemann; Rafal Dziadziuszko; Johan Vansteenkiste; Cathal O'Brien

Introduction: The reported prevalence of ALK receptor tyrosine kinase gene (ALK) rearrangement in NSCLC ranges from 2% to 7%. The primary standard diagnostic method is fluorescence in situ hybridization (FISH). Recently, immunohistochemistry (IHC) has also proved to be a reproducible and sensitive technique. Reverse‐transcriptase polymerase chain reaction (RT‐PCR) has also been advocated, and most recently, the advent of targeted next‐generation sequencing (NGS) for ALK and other fusions has become possible. This study compares anaplastic lymphoma kinase (ALK) evaluation with all four techniques in resected NSCLC from the large European Thoracic Oncology Platform Lungscape cohort. Methods: A total of 96 cases from the European Thoracic Oncology Platform Lungscape iBiobank, with any ALK immunoreactivity were examined by FISH, central RT‐PCR, and NGS. An H‐score higher than 120 defines IHC positivity. RNA was extracted from the same formalin‐fixed, paraffin‐embedded tissues. For RT‐PCR, primers covered the most frequent ALK translocations. For NGS, the Oncomine Solid Tumour Fusion Transcript Kit (Thermo Fisher Scientific, Waltham, MA) was used. The concordance was assessed using the Cohen &kgr; coefficient (two‐sided &agr; ≤ 5%). Results: NGS provided results for 77 of the 95 cases tested (81.1%), whereas RT‐PCR provided results for 77 of 96 (80.2%). Concordance occurred in 55 cases of the 60 cases tested with all four methods (43 ALK negative and 12 ALK positive). Using ALK copositivity for IHC and FISH as the criterion standard, we derived a sensitivity for RT‐PCR/NGS of 70.0%/85.0%, with a specificity of 87.1%/79.0%. When either RT‐PCR or NGS was combined with IHC, the sensitivity remained the same, whereas the specificity increased to 88.7% and 83.9% respectively. Conclusion: NGS evaluation with the Oncomine Solid Tumour Fusion transcript kit and RT‐PCR proved to have high sensitivity and specificity, advocating their use in routine practice. For maximal sensitivity and specificity, ALK status should be assessed by using two techniques and a third one in discordant cases. We therefore propose a customizable testing algorithm. These findings significantly influence existing testing paradigms and have clear clinical and economic impact.


Diagnostic Cytopathology | 2018

The Yokohama system for reporting directly sampled endometrial cytology: The quest to develop a standardized terminology

Franco Fulciniti; Kenji Yanoh; Petros Karakitsos; Jun Watanabe; Alessia Di Lorito; Niki Margari; Yoshinobu Maeda; Maki Kihara; Yoshiaki Norimatsu; Tadao K. Kobayashi; Yasuo Hirai

The main purpose of directly sampled endometrial cytology is to detect invasive endometrial malignancies. With this principle in mind, The Yokohama System (TYS) Working Group, composed of cytopathologists, surgical pathologists, and gynecologic oncologists met at the 2016 International Congress of Cytology, Yokohama, with the aim to publish a standardized reporting system inclusive of specific diagnostic categories and cytomorphologic criteria for uniform and reliable diagnosis of endometrial malignancies on directly sampled endometrial samples.


Acta Cytologica | 2014

Expression of PTEN in endometrial liquid-based cytology.

Alessia Di Lorito; Roberta Zappacosta; Serena Capanna; Daniela Maria Pia Gatta; Sandra Rosini; Fernando Schmitt

Objectives: Endometrial cytology offers a reliable alternative to biopsy in endometrial cancer detection and it may be useful in obtaining material to study prognostic and predictive markers. Over the years, new sampling devices have been developed. Molecular alterations in endometrial cancers were previously described using formalin-fixed paraffin-embedded tissues with particular attention, in endometrioid carcinomas, to the PTEN-PI3K pathway. PTEN evaluation could be useful in endometrial carcinomas for selecting patients for target therapies. Study Design: We studied 51 endometrial samples collected using the Endogyn device and 71 obtained with the Endoflower dispositive device, and processed using liquid-based cytology. Most of the cases were matched with a corresponding histological biopsy. The overall accuracy of Endoflower was 100%. Immunohistochemistry (IHC) and immunocytochemistry (ICC) for PTEN were performed using monoclonal antibody 6H2.1 from DAKO. Results: The IHC showed PTEN-null glands in 4 cases. The same cancers were negative in ICC. Among the 10 carcinomas on cytology, PTEN-null glands were found in 1 case. All the normal endometrium control cases were positive in cytology and histology. Conclusions: Our results suggest that endometrial devices provide useful material for the diagnosis and evaluation of PTEN expression.


Archive | 2018

Molecular Cytology Applications on the Lung

Alessia Di Lorito; Daniel Stieber; Fernando Schmitt

Lung cancer is the leading cause of cancer death in the world. Lung cytopathology is a significant part of the cytopathology practice. Less than 30% of patients with non-small cell lung cancer (NSCLC) are eligible for surgical treatment, and more than 70% of the NSCLCs have only cytological specimens available for molecular analysis. The most frequently used techniques to obtain material are bronchoscopy and fine-needle aspiration (FNA), performed under imaging guidance; EUS-guided FNA is used to stage lung cancer and to study mediastinal lesions. These materials are used to determine the origin and the nature of the lesions and to apply ancillary techniques. The development of molecular heterogeneity in NSCLC has led to the identification of molecular subgroups, which are responsive to target therapies, especially in lung adenocarcinoma that is the most common histological subtype. The clinical relevance of EGFR (epidermal growth factor receptor) and KRAS (Kirsten rat sarcoma viral oncogene) mutational status has been established, and testing these mutations by sequencing or RT-PCR (Reverse transcriptase polymerase chain reaction) methods has become a standard practice. The discovery of ALK (anaplastic lymphoma kinase) translocations and the responsiveness of these tumors to ALK inhibitors have led to study them by FISH (fluorescence in situ hybridization) and RT-PCR. Other genes such as ROS1, RET, and c-Met have been routinely assessed for targeted therapies. Recently the introduction of next-generation sequencing has focused on new gene alterations, although their clinical relevance has not been well established, yet. Recently, gene panels RNA-based to check rearrangements are under evaluation, and probably in future they will be developed also in clinical settings in order to find all molecular alterations.


Journal of Thoracic Disease | 2017

Why anti-PD1/PDL1 therapy is so effective? Another piece in the puzzle

Antonio Marchetti; Alessia Di Lorito; Fiamma Buttitta

Immune checkpoint inhibitors anti-programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) are currently changing the approach of treatment of non-small cell lung cancer patients (NSCLCs). During the last two years, the anti-PD-1 inhibitors, nivolumab (OPDIVO, Bristol-Myers Squibb) and pembrolizumab (KEYTRUDA, Merck Sharp and Dohme Corporation) and the anti-PD-L1 inhibitor atezolizumab (TECENTRIQ, Genentech Oncology) have been approved by the U.S. Food and Drug Administration (FDA) in the treatment of patients with advanced NSCLC with progression on or after first-line therapy. Indeed, the European Medicines Agency (EMA) has endorsed Nivolumab and Pembrolizumab for the same indication. In recent times, pembrolizumab has also been recommended by both the U.S. and European agencies for the first-line therapy of NSCLCs with advanced disease. Furthermore, two other drugs as durvalumab (MEDI4736, AstraZeneca) and avelumab (MSB0010718C, Merck KGaA & Pfizer) are being examined for the treatment of NSCLC patients (1).


JCO Precision Oncology | 2017

ROS1 Gene Fusion in Advanced Lung Cancer in Women: A Systematic Analysis, Review of the Literature, and Diagnostic Algorithm

Antonio Marchetti; Massimo Barberis; Alessia Di Lorito; Maria Vittoria Pace; Chiara Di Lisio; Lara Felicioni; Elena Guerini-Rocco; Andrea Vingiani; Tommaso D’Antuono; Marcella Liberatore; Giampaolo Filice; Graziano De Luca; Filippo De Marinis; Antonio Passaro; L. Guetti; Luciana Irtelli; Lucio Crinò; Felice Mucilli; Fiamma Buttitta

PurposeCrizotinib, a mesenchymal-epithelial transition/anaplastic lymphoma kinase/c-ros oncogene 1 (ROS1) inhibitor, has recently been approved by the US Food and Drug Administration for the treatment of patients with advanced ROS1-positive non–small-cell lung cancer (NSCLC). Therefore, interest in ROS1 testing is growing. ROS1 gene fusions affect approximately 0.5% to 2% of unselected NSCLCs. Limited data are available on the prevalence and distribution of ROS1 fusions in patients with advanced-stage NSCLC.Material and MethodsA series of 727 lung adenocarcinomas from patients with stage IV disease, negative for epidermal growth factor receptor and anaplastic lymphoma kinase alterations, were tested for ROS1 fusions by fluorescent in situ hybridization analysis, with confirmation by immunohistochemistry. Results were correlated with clinicopathologic parameters and compared with data from the literature.ResultsROS1 fusions were detected in 29 patients (4%), including 27 of 266 females (10.2%) and two of 4...

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Giampaolo Filice

University of Chieti-Pescara

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L. Guetti

University of Chieti-Pescara

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Maria Vittoria Pace

University of Chieti-Pescara

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Roberta Zappacosta

University of Chieti-Pescara

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Sandra Rosini

University of Chieti-Pescara

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Tommaso D’Antuono

University of Chieti-Pescara

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