Poster Presentation 38th Lorne Cancer Conference 2026

50-marker multiplexed imaging transforms the potential of circulating tumor cells to guide precision oncology (#211)

Timothy J Mann 1 , Ye Zheng 1 , Tanzila Khan 2 3 , Tim Huang 4 , Udit Nindra 3 5 , Yafeng Ma 2 3 , Alexander James 2 6 , Daniel P Neumann 1 , Felix V Kohane 1 , Ihuan Gunawan 1 , Fatemeh Vafaee 4 7 8 , Wei Chua 3 5 , Paul de Souza 5 , Tara L Roberts 2 3 , Therese M Becker 2 3 6 , John G Lock 1 2 7
  1. School of Biomedical Sciences, Faculty of Medicine and Health, University of New South Wales, Sydney, NSW, Australia
  2. Ingham Institute for Applied Medical Research, Liverpool, NSW, Australia
  3. School of Medicine, Western Sydney University, Sydney, NSW, Australia
  4. OmniOmics.AI , Sydney, NSW, Australia
  5. Depatment of Medical Oncology, Liverpool Hospital, Liverpool, NSW, Australia
  6. South Westerns Sydney Clinical School, University of New South Wales, Sydney, NSW, Australia
  7. Artificial Intelligence Institute, University of New South Wales, Sydney, NSW, Australia
  8. School of Biotechnology & Biomolecular Sciences, Faculty of Science, University of New South Wales, Sydney, NSW, Australia

Liquid biopsy-derived circulating tumour cells (CTCs) could greatly potentiate precision oncology by yielding actionable biomarkers spanning molecular (oncogenic/resistance) signals to cancer cell-phenotypes; all via minimally invasive sampling that is compatible with longitudinal patient monitoring (unlike solid biopsy). Yet typical CTC profiling via standard 4-5 marker immunofluorescence imaging provides insufficient molecular bandwidth to guide precision oncology given adaptable, diversifying disease-states and an ever-growing therapeutic arsenal. To achieve a step-change in CTC utility as molecular guides for precision oncology, we have increased CTC profiling depth at least ten-fold via an end-to-end pipeline for deep multiplexed imaging, capturing 50 molecular markers per CTC. We quantify the expression, phosphorylation and subcellular localization of known and putative biomarkers that: define CTCs and cancer phenotypes; directly read-out oncogenic/resistance signalling, and/or; are explicit therapeutic targets. Validated via detection of known resistance-biomarkers in a three-stage resistance-progression model of prostate cancer, translational analysis of prostate cancer patient-derived CTCs then confirmed that our deep multiplexed profiling: i) improves CTC classification and; ii) captures inter- and intra-patient CTC heterogeneity corresponding to therapy responses. Machine learning then identified; iii) therapeutically actionable profiles per patient, integrating molecular expression and subcellular localization. This demonstrates proof-of-principle capacity for deep multiplexed CTC image-profiling to derive unprecedented molecular and cellular insights suited to guiding precision oncology.