Accurate Classification of Non-small Cell Lung Cancer (NSCLC) Pathology and Mapping of EGFR Mutation Spatial Distribution by Ambient Mass Spectrometry Imaging

Abstract

Objectives: Tumor pathology examination especially epidermal growth factor receptor ( EGFR ) mutations molecular testing has been integral part of lung cancer clinical practices. However, the EGFR mutations spatial distribution characteristics remains poorly investigated, which is critical to tumor heterogeneity analysis and precision diagnosis. Here, we conducted an exploratory study for label-free lung cancer pathology diagnosis and mapping of EGFR mutation spatial distribution using ambient mass spectrometry imaging (MSI). Materials and Methods: MSI analysis were performed in 55 post-operative non-small cell lung cancer (NSCLC) tumor and paired normal tissues to distinguish tumor from normal and classify pathology. We then compared diagnostic sensitivity of MSI and ADx-amplification refractory mutation system (ARMS) for the detection of EGFR mutation in pathological confirmed lung adenocarcinoma (AC) and explored EGFR mutations associated biomarkers to depict EGFR spatial distribution base on ambient MSI. Results: Of 55 pathological confirmed NSCLC, MSI achieved a diagnostic sensitivity of 85.2% (23/27) and 82.1% (23/28) for AC and squamous cell carcinoma (SCC), respectively. Among 27 AC, there were 17 EGFR- wild-type and 10 EGFR- mutated-positive samples detected by ARMS, and MSI achieved a diagnostic sensitivity of 82.3% (14/17) and 80% (8/10) for these two groups. Several phospholipids were specially enriched in AC compared with SCC tissues, with the higher ions intensity of phospholipids in EGFR- mutated-positive compared with EGFR- wild-type AC tissues. We also found EGFR mutations distribution was heterogeneous in different regions of same tumor by multi-regions ARMS detection, and only the regions with higher ions intensity of phospholipids were EGFR- mutated-positive. Conclusion: MSI method could accurately distinguish tumor pathology and subtypes, and phospholipids were reliable EGFR mutations associated biomarkers, phospholipids imaging could intuitively visualize EGFR mutations spatial distribution, may facilitate our understanding of tumor heterogeneity.

Advances of single-cell genomics and epigenomics in human disease: where are we now?

“…This is especially important when only a specific part of the tumour is targeted and proliferation is still possible even from a minute proportion of cells, often concealed in bulk analyses (Qian et al 2017 ; Tirosh and Suvà 2019 ). For example, in studies investigating lung adenocarcinomas, accurate profiling of tumours for targeted treatment remains challenging due to increased tumour heterogeneity (Zhang et al 2019 ). While mutations of the epidermal growth factor receptor ( EFGR ) in non-small cell lung cancers (NSCLC) have been widely employed as biomarkers for lung carcinogenesis (Harrison et al 2019 ; Zhang et al 2019 ), resistance to the well-established EGFR tyrosine kinase inhibitors has been frequently reported, most likely due to EGFR T790M resistance mutations (Del Re et al 2018 ; Rexer et al 2009 ; Sullivan and Planchard 2016 ).…”

The Co-Expression of Programmed Death-Ligand 1 (PD-L1) in Untreated EGFR-Mutated Metastatic Lung Adenocarcinoma

“…Advanced and surgically unresectable lung cancer is the leading cause of cancer-related mortality in both men and women worldwide [1]. According to the classification of histopathology, non-small-cell lung cancer (NSCLC) accounts for 85% of cases, and adenocarcinoma is the most common histological type of NSCLC (~50%) [2,3]. Active mutation in the epidermal growth factor receptor (EGFR) kinase domain has been well studied and is known to be the main oncogenic-driven mutation in lung adenocarcinoma [4,5].…”

Role of SOX Protein Groups F and H in Lung Cancer Progression

“…Moreover, LCINS is associated with a range of characteristic mutations different from those present in smokers [19]. In the histological classification, two main groups of lung cancer can be found: small cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC) [20][21][22]. This distribution has an important clinical significance since these tumors are not only different in terms of their histogeneses, histological structures and disease courses, but also with regard to their methods of treatment and response to the therapies applied [21,22].…”

“…NSCLCs are much more common than SCLCs and represent approximately 85% of all lung cancers. Over half of these cancers are detected at an advanced clinical stage-i.e., III or IV [20][21][22][23][24][25][26][27]. NSCLCs are more diverse than SCLCs and are less responsive to common chemotherapy [28].…”

“…It has been very well documented that in the case of NSCLC progression, most mutations are associated with pathways related to the activation of the epidermal growth factor receptor protein (EGFR)-i.e., RAS/RAF/MEK/ERK and PI3K, as well as AKT/mTOR. Changes in the EGFR pathways lead to an increase in cancer cell proliferation and their ability to metastasize and initiate the angiogenesis process [20,22,26,36]. If one compares normal lung tissue and small-lung cell lung cancer, the EGFR membrane expression is more frequent in NSCLC cases-it has been observed in 40-80% cases of NSCLC, including 24-89% of LSCCs and 23-46% of ACs [37][38][39][40].…”