Abstract 5308: Analysis of extracellular vesicle surface markers as small cell lung cancer biomarkers
Authors: Ozawa Patricia M. , Groves Sarah M. , Quaranta Vito , Weaver Alissa M. 
Published: June 2023

Small Cell Lung Cancer (SCLC) is an aggressive tumor type, usually metastatic at diagnosis, with a median survival of less than a year. SCLC tumors are heterogeneous and are composed of neuroendocrine (A, A2, N) and non-neuroendocrine (Y) subtypes, with the cooperation by heterogeneous cells being key to drive tumor survival. Unfortunately, the current SCLC treatment approach does not take tumor heterogeneity into consideration. Importantly, there are no consensus markers that can identify SCLC subtypes or early tumors. Extracellular vesicles (EVs), small vesicles known to promote cell to cell communication, have been investigated as non-invasive biomarkers for cancer diagnostics, staging, and treatment monitoring. EVs are attractive candidates for biomarker research due to the selective packaging of RNA, DNA, and proteins that are then secreted in the blood. Therefore, our aim was to identify new biomarkers based on EV surface proteins with potential for early detection and tumor characterization. We used differential ultracentrifugation to isolate EVs from four SCLC subtypes A, A2, N, and Y, which were submitted to label-free mass spectrometry (EV-proteomics). We identified a total of 3011 proteins, with the two most distinct subtypes being A2 and N, with 786 differentially expressed EV-proteins. Meanwhile, the 2 most similar subtypes were N and A, with only 75 differentially expressed EV-proteins. Gene set enrichment analysis of EVs derived from the A2 subtype showed an association with pathways in cancer, chemokine signaling, and neuronal systems. In addition, PANTHER analysis identified enrichment in nicotine pharmacodynamic pathways on these EVs, which corroborate the association of A2 cell lines with drug resistance. EVs derived from the N and A subtypes were enriched in spliceosome, RNA processing, and metabolism, while EVs from Y cell lines were enriched in extracellular matrix organization and metabolic pathways, similar to pathways from cell line RNAseq data. 13 proteins were differentially expressed in all group comparisons, including proteins involved in PI3K and nicotinic acetylcholine receptor signaling pathways. Further comparison of EV-proteomics with top genes from CIBERSORT analysis using cell line RNAseq data showed an overlap of 9 surface proteins, with 3 possible SCLC pan markers. In conclusion, EV-proteins isolated from SCLC cell lines presented a unique signature for each subtype, especially EVs derived from A2 subtype, which has a signature associated with drug resistance. We identified surface proteins on SCLC EVs with potential as SCLC pan and tumor characterization, which will be validated on EVs derived from cell lines and patient plasma by Western blotting and single-EV microflow cytometry.

Citation Format: Patricia M. Ozawa, Sarah M. Groves, Vito Quaranta, Alissa M. Weaver. Analysis of extracellular vesicle surface markers as small cell lung cancer biomarkers. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5308.