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43 Highly multiplexed digital spatial profiling of the tumor microenvironment of non-small-cell lung cancer (NSCLC)
  1. Arutha Kulasinghe1,
  2. Connor O’Leary2,
  3. Rahul Ladwa2,
  4. Majid Warkiani3 and
  5. Kenneth O’byrne1
  1. 1Queensland University of Technology, Kelvin Grove, Australia
  2. 2Princess Alexandra Hospital, Woolloongabba, Australia
  3. 3University of Technology Sydney, Sydney, Australia


Background 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.

Methods In this study, we utilised the Nanostring GeoMX Digital Spatial Profiler (DSP) platform to profile a NSCLC tissue microarray for protein markers across immune cell profiling, immuno-oncology(IO) drug target, 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.

Results Our data revealed that paired analysis (n=18) of patient matched compartments indicated that the TME was significantly enriched in CD27, CD3, CD4, CD44, CD45, CD45RO, CD68, CD163, and VISTA relative to tumour. Unmatched analysis indicated that the NAT(n=19) was significantly enriched in CD34, fibronectin, IDO1, LAG3, ARG1 and PTEN when compared to the TM E(n=32). Univariate Cox proportional hazards indicated that the presence of cells expressing CD3 (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).

Conclusions 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.

Acknowledgements This study was funded by the Princess Alexandra Hospital Foundation grant for KOB. AK is supported by an NHMRC ECF Fellowship (APP1157741) and Cure Cancer (APP1182179).

Ethics Approval The study was approved by the QUT Human Research Ethics Board

This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See:

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