High-Plex and High-Throughput Digital Spatial Profiling of Non-Small-Cell Lung Cancer (NSCLC)

James Monkman; Touraj Taheri; Majid Ebrahimi Warkiani; Connor O'Leary; Rahul Ladwa; Derek Richard; Ken O'Byrne; Arutha Kulasinghe

doi:10.3390/cancers12123551

High-Plex and High-Throughput Digital Spatial Profiling of Non-Small-Cell Lung Cancer (NSCLC)

Cancers (Basel). 2020 Nov 27;12(12):3551. doi: 10.3390/cancers12123551.

Authors

James Monkman^{1

2

3}, Touraj Taheri^{4

5}, Majid Ebrahimi Warkiani⁶, Connor O'Leary^{1

2

3

7}, Rahul Ladwa^{5

7}, Derek Richard^{1

2

3}, Ken O'Byrne^{1

2

3

7}, Arutha Kulasinghe^{1

2

3

8}

Affiliations

¹ School of Biomedical Sciences, Faculty of Health and Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, QLD 4000, Australia.
² Translational Research Institute, Woolloongabba, QLD 4102, Australia.
³ Cancer and Ageing Research Program, Translational Research Institute, Brisbane, QLD 4000, Australia.
⁴ Queensland Pathology, Herston, QLD 4006, Australia.
⁵ School of Medicine, University of Queensland, Brisbane, QLD 4102, Australia.
⁶ School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia.
⁷ Princess Alexandra Hospital, Woolloongabba, QLD 4102, Australia.
⁸ Institute of Molecular Bioscience, University of Queensland, Brisbane, QLD 4072, Australia.

Abstract

Profiling the tumour microenvironment (TME) has been informative in understanding the underlying tumour-immune interactions. Multiplex immunohistochemistry (mIHC) coupled with molecular barcoding technologies have revealed greater insights into the TME. In this study, we utilised the Nanostring GeoMX Digital Spatial Profiler (DSP) platform to profile a non-small-cell lung cancer (NSCLC) tissue microarray for protein markers across immune cell profiling, immuno-oncology (IO) drug targets, immune activation status, immune cell typing, and pan-tumour protein modules. Regions of interest (ROIs) were selected that described tumour, TME, and normal adjacent tissue (NAT) compartments. Our data revealed that paired analysis (n = 18) of matched patient compartments indicate that the TME was significantly enriched in CD27, CD3, CD4, CD44, CD45, CD45RO, CD68, CD163, and VISTA relative to the tumour. Unmatched analysis indicated that the NAT (n = 19) was significantly enriched in CD34, fibronectin, IDO1, LAG3, ARG1, and PTEN when compared to the TME (n = 32). Univariate Cox proportional hazards indicated that the presence of cells expressing CD3 (hazard ratio (HR): 0.5, p = 0.018), CD34 (HR: 0.53, p = 0.004), and ICOS (HR: 0.6, p = 0.047) in tumour compartments were significantly associated with improved overall survival (OS). We implemented both high-plex and high-throughput methodologies to the discovery of protein biomarkers and molecular phenotypes within biopsy samples, and demonstrate the power of such tools for a new generation of pathology research.

Keywords: NSCLC; differential protein expression; nanostring GeoMx digital spatial profiling; spatial; tissue microarray; tumour microenvironment.