Faculty Bibliography

The clinical use of blood tests for the detection of malignant pleural mes-othelioma (MPM) or patient's stratification is currently hampered by the lack of sensitive blood biomarkers. In a previous work, we have identified a targeted proteomics candidate biomarkers signature for MPM from the blood and composed of seven N-linked glycopeptides (N-glycopeptides). Here, we have verified a reduced version of the original proteomics sig-nature (now composed of six N-glycopeptides) within a multicenter co-hort of more than 400 serum samples collected in the USA, Australia and Europe from MPM patients and cancer-free asbestos exposed donors. The AUC of the proteomics signature was 0.738 for discriminating MPM patients and asbestos exposed donors. For early stage MPM (stage I/II) AUC was 0.765. These results were in line with what we observed by using an FDA approved ELISA test for the best investigated MPM blood biomarker SMRP (soluble mesothelin-related peptides). This highlighted the potential of our academically developed MPM proteomics signature once further optimized for routine clinical use. The negative likelihood ratio of the proteomics signature was 0.11 for early stage MPM, reflecting its higher sensitivity also if compared to SMRP. This will be relevant in the context of early disease detection and screenings for thoracic malig-nancies. Further, the proteomics signature demonstrated prognostic ca-pacity, separating high-risk and low-risk groups of MPM patients with a hazard ratio of 1.659 and which may support clinical treatment decisions. In summary, targeted proteomics permitted to establish a blood based biomarkers signature with diagnostic and prognostic ability for MPM

Introduction/UNASSIGNED:Most diseases involve a complex interplay between multiple biological processes at the cellular, tissue, organ, and systemic levels. Clinical tests and biomarkers based on the measurement of a single or few analytes may not be able to capture the complexity of a patient's disease. Novel approaches for comprehensively assessing biological processes from easily obtained samples could help in the monitoring, treatment, and understanding of many conditions. Objectives/UNASSIGNED:We propose a method of creating scores associated with specific biological processes from mass spectral analysis of serum samples. Methods/UNASSIGNED:A score for a process of interest is created by: (i) identifying mass spectral features associated with the process using set enrichment analysis methods, and (ii) combining these features into a score using a principal component analysis-based approach. We investigate the creation of scores using cohorts of patients with non-small cell lung cancer, melanoma, and ovarian cancer. Since the circulating proteome is amenable to the study of immune responses, which play a critical role in cancer development and progression, we focus on functions related to the host response to disease. Results/UNASSIGNED:We demonstrate the feasibility of generating scores, their reproducibility, and their associations with clinical outcomes. Once the scores are constructed, only 3 µL of serum is required for the assessment of multiple biological functions from the circulating proteome. Conclusion/UNASSIGNED:These mass spectrometry-based scores could be useful for future multivariate biomarker or test development studies for informing treatment, disease monitoring and improving understanding of the roles of various biological functions in multiple disease settings.

Asbestos causes malignant transformation of primary human mesothelial cells (HM), leading to mesothelioma. The mechanisms of asbestos carcinogenesis remain enigmatic, as exposure to asbestos induces HM death. However, some asbestos-exposed HM escape cell death, accumulate DNA damage, and may become transformed. We previously demonstrated that, upon asbestos exposure, HM and reactive macrophages releases the high mobility group box 1 (HMGB1) protein that becomes detectable in the tissues near asbestos deposits where HMGB1 triggers chronic inflammation. HMGB1 is also detectable in the sera of asbestos-exposed individuals and mice. Searching for additional biomarkers, we found higher levels of the autophagy marker ATG5 in sera from asbestos-exposed individuals compared to unexposed controls. As we investigated the mechanisms underlying this finding, we discovered that the release of HMGB1 upon asbestos exposure promoted autophagy, allowing a higher fraction of HM to survive asbestos exposure. HMGB1 silencing inhibited autophagy and increased asbestos-induced HM death, thereby decreasing asbestos-induced HM transformation. We demonstrate that autophagy was induced by the cytoplasmic and extracellular fractions of HMGB1 via the engagement of the RAGE receptor and Beclin 1 pathway, while nuclear HMGB1 did not participate in this process. We validated our findings in a novel unique mesothelial conditional HMGB1-knockout (HMGB1-cKO) mouse model. Compared to HMGB1 wild-type mice, mesothelial cells from HMGB1-cKO mice showed significantly reduced autophagy and increased cell death. Autophagy inhibitors chloroquine and desmethylclomipramine increased cell death and reduced asbestos-driven foci formation. In summary, HMGB1 released upon asbestos exposure induces autophagy, promoting HM survival and malignant transformation.

BACKGROUND:We have verified a mass spectrometry (MS)-based targeted proteomics signature for the detection of malignant pleural mesothelioma (MPM) from the blood. METHODS:A seven-peptide biomarker MPM signature by targeted proteomics in serum was identified in a previous independent study. Here, we have verified the predictive accuracy of a reduced version of that signature, now composed of six-peptide biomarkers. We have applied liquid chromatography-selected reaction monitoring (LC-SRM), also known as multiple-reaction monitoring (MRM), for the investigation of 402 serum samples from 213 patients with MPM and 189 cancer-free asbestos-exposed donors from the United States, Australia, and Europe. RESULTS:= 0.021). CONCLUSIONS:Targeted proteomics allowed the development of a multianalyte signature with diagnostic and prognostic potential for MPM from the blood. IMPACT/CONCLUSIONS:The proteomic signature represents an additional diagnostic approach for informing clinical decisions for patients at risk for MPM.

INTRODUCTION/BACKGROUND:A grading system for pulmonary adenocarcinoma has not been established. The IASLC pathology panel evaluated a set of histological criteria associated with prognosis aimed at establishing a grading system for invasive pulmonary adenocarcinoma. DESIGN/METHODS:A multi-institutional study involving multiple cohorts of invasive pulmonary adenocarcinomas was conducted. A cohort of 284 stage I pulmonary adenocarcinomas was used as a training set to identify histological features associated with patient outcomes (recurrence-free survival [RFS] and overall survival [OS]). ROC curve analysis was used to select the best model, which was validated (n=212) and tested (n=300, including Stage I-III) in independent cohorts. Reproducibility of the model was assessed using kappa statistics. RESULTS:The best model (AUC= 0.749 for RFS and 0.787 for OS) was composed of a combination of predominant plus high-grade histological pattern with a cut off of 20% for the latter. The model consists of: Grade 1: lepidic predominant tumor; Grade 2: acinar or papillary predominant tumor, both with no or less than 20% of high-grade patterns; and Grade 3: any tumor with 20% or more of high-grade patterns (solid, micropapillary and or complex gland). Similar results were seen in the validation (AUC = 0.732 for RFS and 0.787 for OS) and test cohorts (AUC of 0.690 for RFS and 0.743 for OS), confirming the predictive value of the model. Inter-observer reproducibility showed good agreement (k=0.617). CONCLUSION/CONCLUSIONS:A grading system based on the predominant and high-grade patterns is practical and prognostic for invasive pulmonary adenocarcinoma. ACKNOWLEDGEMENT/UNASSIGNED:This project was supported by grants from the International Association for the study of Lung Cancer (IASLC), and NIH/NCI Early Detection Research Network 1U01CA214195-01 to HP used for biospecimen collection and salary support (DFL). We would like to thank the staff of the Center of Biospecimen Research and Development (CBRD) from the New York University for their help in digital pathology and histology for this project. CBRD is partially supported by the Cancer Center Support Grant P30CA016087 at the Laura and Isaac Perlmutter Cancer Center.

Cell division and organismal development are exquisitely orchestrated and regulated processes. The dysregulation of the molecular mechanisms underlying these processes may cause cancer, a consequence of cell-intrinsic and/or cell-extrinsic events. Cellular DNA can be damaged by spontaneous hydrolysis, reactive oxygen species, aberrant cellular metabolism or other perturbations that cause DNA damage. Moreover, several environmental factors may damage the DNA, alter cellular metabolism or affect the ability of cells to interact with their microenvironment. While some environmental factors are well established as carcinogens, there remains a large knowledge gap of others owing to the difficulty in identifying them because of the typically long interval between carcinogen exposure and cancer diagnosis. DNA damage increases in cells harbouring mutations that impair their ability to correctly repair the DNA. Tumour predisposition syndromes in which cancers arise at an accelerated rate and in different organs - the equivalent of a sensitized background - provide a unique opportunity to examine how gene-environment interactions influence cancer risk when the initiating genetic defect responsible for malignancy is known. Understanding the molecular processes that are altered by specific germline mutations, environmental exposures and related mechanisms that promote cancer will allow the design of novel and effective preventive and therapeutic strategies.

Among more than 200 BAP1-mutant families affected by the "BAP1 cancer syndrome," nearly all individuals inheriting a BAP1 mutant allele developed one or more malignancies during their lifetime, mostly uveal and cutaneous melanoma, mesothelioma, and clear-cell renal cell carcinoma. These cancer types are also those that, when they occur sporadically, are more likely to carry somatic biallelic BAP1 mutations. Mechanistic studies revealed that the tumor suppressor function of BAP1 is linked to its dual activity in the nucleus, where it is implicated in a variety of processes including DNA repair and transcription, and in the cytoplasm, where it regulates cell death and mitochondrial metabolism. BAP1 activity in tumor suppression is cell type- and context-dependent. BAP1 has emerged as a critical tumor suppressor across multiple cancer types, predisposing to tumor development when mutated in the germline as well as somatically. Moreover, BAP1 has emerged as a key regulator of gene-environment interaction.This article is highlighted in the In This Issue feature, p. 1079.

BACKGROUND:Our objective is to report our outcomes and demonstrate our evolving technique for robotic sleeve resection of the airway, with or without lobectomy, using video vignettes. METHODS:We retrospectively reviewed a single surgeon prospective database from October 2010 to October 2019. RESULTS:Over 9 years, there were 5,573 operations of which 1951 were planned for a robotic approach. There were 755 robotic lobectomies, 306 robotic segmentectomies, and 23 consecutive patients were scheduled for elective completely portal, robotic sleeve resection. Sleeve lobectomy was performed in 18 patients: 10 right upper lobe, 6 left upper lobe, and 2 right lower lobe. Two patients had mainstem bronchus resections and two underwent right bronchus intermedius resections that preserved all of the lung. One patient had a robotic pneumonectomy. There was one conversion to open thoracotomy due to concern for anastomotic tension in a patient who received neoadjuvant therapy. All patients had an R0 resection. In the last 10 operations, we modified our airway anastomosis, using a running self-locking absorbable suture. The median length of stay was 3 days (range 1-11). There were no 30- or 90-day mortalities. Within a median follow-up of 18 months, there were no anastomotic strictures and no recurrent cancers. CONCLUSIONS:Our early and midterm results show that a completely portal robotic sleeve resection is safe and oncologically effective. The technical aspects of a robotic sleeve resection of the airway are demonstrated using video vignettes.