Recent screening trial results indicate that low-dose computed tomography (LDCT) reduces lung cancer mortality in high-risk patients. However, high false-positive rates, costs, and potential harms highlight the need for complementary biomarkers.

The bioMILD trial is designed to test the efficiency of a combined MSC biomarker-LDCT imaging approach as first-line screening tests in 4000 heavy smokers 50 years or older; results will be investigated in association with smoking habits, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis and C-reactive protein levels in plasma. Moreover, MSC will be implemented by integrating determination of additional biomarkers in plasma samples from the individuals in the bioMILD trial:

2
Peripheral blood immune cell profile

3
Mutation profile of circulating tumor DNA

1
Evaluation of HGF and ECM-related molecules

publications

Group leaders

Gabriella Sozzi

Ugo Pastorino

Group presentation

The Tumor Genomics Unit is focused on the study of the molecular pathogenesis of lung cancer. Our research activity is centered on all aspects of lung cancer with the final aim of making an impact on a disease that represents a major health-care burden in terms of morbidity and mortality with 5-year survival estimates around 15%. We use an integrated approach that combines cellular and molecular biology, biochemistry and pharmacology to gain new insights in the pathogenesis of lung cancer and on novel ways to provide early diagnosis and novel treatment options. The research program includes the analysis of (epi)genetic changes, phenotypic analyses and functional studies aimed at the elucidation of the contribution of the tumor and its microenvironment in the carcinogenetic process. The goal of our translational studies is the implementation of highly sensitive molecular tests that could be used within screening programs to improve both detection and clinical management of lung cancers. In the field of circulating biomarkers, we have a demonstrated record of successful and productive research projects in an area such as the screening for Lung Cancer of high relevance for the National Health System of almost every country.

Our studies are supported by AIRC (Investigator and 5X1000 Special Program), Ministry of Health and National Cancer Institute (USA) grants.

[Our early achievements include the identification and characterization of the key genetic events occurring in the lung carcinogenetic process through the cytogenetic and molecular analysis of normal bronchial tissue, precancerous lesions and invasive cancer. We have contributed to the identification and characterization of the functional role of the oncosuppressor gene FHIT in lung carcinogenesis. In the field of biomarkers we have developed sensitive methods and patents for the evaluation of free circulating DNA in plasma for diagnostic purposes. Our work has also led to the identification of cancer initiating cells in lung cancer and the characterization of microRNA aberrant expression in lung tumor and normal tissues and their functional and therapeutic studies. For the therapeutic approaches original patients-derived xenografts (PDXs), in mouse models are established. We have recently identified a circulating plasma microRNA signature which led to the development and patent of a three level risk classifier (MSC test) based on 24 plasma miRNAs with predictive, diagnostic, and prognostic potential]

The two GLs, Dr. Pastorino, a thoracic surgeon with solid expertise in lung cancer screening trials, and a geneticist, Dr. Sozzi, with a strong background in lung cancer biology, genetics, and biomarkers, have complementary experience and will cooperate to run the study. Based on their previous studies, they launched a prospective screening trial, bioMILD, on 4,000 volunteers to test the performance of a 3-level risk classifier (MSC High, Intermediate, Low) based on 24 plasma miRNAs in conjunction with low-dose computed tomography (LDCT). The latter is an original and unique screening trial combining molecular and a radiological biomarkers that collects large amounts of biological samples which will be useful for validation of an enlarged number of different biomarkers and could be made also available to the scientific community for testing the most promising biomarkers generated from other studies.

In previous studies, we identified a 3-level risk classifier (MSC High, H; Intermediate, I; Low, L) based on 24 plasma miRNAs with predictive, diagnostic, and prognostic potential in subjects enrolled in screening trials for lung cancer with LDCT. Analyses of miRNA expression in cells and conditioned medium of various cell types and miRNA in situ hybridization of lung tissues proved a stromal and immune cells-related origin of most miRNAs composing the classifier supporting the utility of biomarkers generated by the dynamic interaction of cancer cells with the microenvironment and the host (immune) response for risk assessment and early detection of lung cancer.

 

Role in the project, experimental design

The excellent clinical performance of the MSC test in the context of lung cancer screening programs has already been validated in 2 datasets of prospectively collected and retrospectively analyzed samples. Since the final phase of biomarkers development is to prove their utility in prospective studies, in 2013 we launched a unique prospective trial, bioMILD (www.bioMILD.org), to test the efficiency of a combined MSC biomarker-LDCT imaging approach as first-line screening tests in 4000 heavy smokers 50 years or older. In the study design, the combination of the results of these 2 tests determines the subsequent diagnostic route as exemplified in the flowchart. As of January 2016, we successfully completed the accrual and executed baseline LDCT-MSC analyses of 4119 subjects. However, prospective screening studies need prolonged follow-up to reach the significant number of events and observation time of subjects in relation with their LDCT imaging and MSC biomarker profile. Therefore, a significant time point will be reached in 2018, with a minimum follow-up time of 3 years. This extension of the project will allow us to reach this end point. Moreover, thanks to reviewers’ request, AIRC invited us to extend the accrual to strengthen the statistical power of the study. We will thus enroll additional 2000 heavy current smokers (≥ 30 p/y)  having blood levels of C-reactive protein (CRP)>2mg/L who represent an high risk group for lung cancer development thus increasing about twice the amount of expected number of events.

During the project we will exploit LDCT and MSC tests with the clinical aim to improve lung cancer screening by combining LDCT and MSC. Major clinical end points are: i) evaluation of sensitivity/specificity/negative predictive value (NPV)/positive predictive value (PPV)/false-positive rate of MSC alone, LDCT alone, and their combination; ii) validation of MSC prognostic value in a prospective setting; iii) assessment of MSC risk profile modulation during follow-up according to LDCT findings; iv) analysis of MSC relationship with smoking habits, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis conditions, and C-reactive protein levels in plasma.

In addition we are also interested in the Improvement and implementation of MSC test. In fact, full clinical validation of MSC as a companion diagnostic tool in a prospective clinical trial is ongoing in the present project. However, the clinical value of the MSC test is currently limited by the number of failed tests due to hemolysis (10%-13%) and the number of false-positives (test specificity ~80%) and false-negatives (test sensitivity ~85%). In this project we will bridge the gap to full clinical validation by integrating determination of additional biomarkers in plasma samples from the individuals of the bioMILD trial.

 

 

 

 

 

 

 

 

 

In particular we will:

  • evaluate circulating ECM-related molecules and HGF as biomarkers for risk assessment. The ECM-HGF molecules will be exploited by commercially available ELISA immunoassay kits. Performance of the markers will be evaluated in high-risk individuals of the bioMILD screening population in relation to the benchmarks represented by LDCT algorithm and MSC. Correlation of the markers with lung function parameters (e.g., FEV1) or diseases (emphysema, COPD) will also be analyzed to investigate potential interactions or value of the different markers in specific clinical settings.
  • combine tumor-derived and microenvironment-derived markers. We reasoned that a circulating tumor DNA (ctDNA)-based assay measuring the tumor-related contribution could possibly implement and add clinical value to the MSC test. In this project, ctDNA mutational load in plasma will be analyzed using a targeted mutation NGS approach and will be applied in those LDCT-positive subjects (nodule >260 mm3) where hemolysis precludes MSC profile and in all false-negative cases of the bioMILD trial.
  • evaluate the peripheral blood immune cell profile as biomarkers for risk assessment. The role of immunity in modulating the risk of disease development remains to be elucidated, while it could have enormous impact in terms of prevention and early intervention. The known immunosuppressive and inflammatory background of lung cancer led us to hypothesize that peripheral blood immune cell profile could be source of biomarkers for risk assessment. We plan to test a peripheral blood mononuclear cell immunosuppressive MIS in the bioMILD trial to assess whether this parameter could help identify high-risk subjects. By taking advantage of color flow cytometry, immunologic markers related to myeloid lineage and T-cell anergy/exhaustion will be included to capture early disease-related immunologic alterations. The MIS will be assessed in all subjects with lung cancer detected in the bioMILD trial including the false-negatives (i.e., MSC low-risk profile) and a paired number of matched disease-free volunteers with MSC high- or low-risk profile. The final aim is to determine if an immunosuppressive index score can implement the accuracy of the miRNA-based MSC test.

Staff

 

Gabriella Sozzi

Biologist, PhD, Group Leader

 

Ugo Pastorino

Thoracic Surgeon, Group Leader

 

Stefano Sestini

Thoracic Surgeon

 

Claudio Jacomelli

Data manager

 

Carla Verri

Biologist

 

Massimo Moro

Biologist

 

Orazio Fortunato

Medical Biotechnologist

 

Mattia Boeri

Medical Biotechnologist, Fellow Fondazione Pezcoller

 

Cristina Borzi

Medical Biotechnologist, junior Fellow

 

Davide Conte

Lab Technician

 

Mavis Mensah

Lab Technician

 

Alfonso Marchianò

Radiologist

 

Giovanni Leuzzi

Thoracic Surgeon

 

Paolo Girotti

Thoracic Surgeon

 

Barbara Sartorelli

Research Nurse

 

Elena Bertocchi

Project/Data Manager

 

Cristina Zanini

Administrative Assistant

 

Copyright © 2017 INT

Fondazione IRCCS Istituto Nazionale dei Tumori

All rights reserved

website by Studio Luvié

Recent screening trial results indicate that low-dose computed tomography (LDCT) reduces lung cancer mortality in high-risk patients. However, high false-positive rates, costs, and potential harms highlight the need for complementary biomarkers.

Copyright © 2017 INT

Fondazione IRCCS Istituto Nazionale dei Tumori

Alla rights reserved

website by Studio Luvié

Project funded with

support from Airc 5x1000

Copyright © 2017 INT

Fondazione IRCCS Istituto Nazionale dei Tumori

All rights reserved

website by Studio Luvié