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Abstract PO-084: Protein marker heterogeneity in breast cancer subtypes measured using immunofluorescence protein multiplexing and quantitative, single cell image analysis

2020· article· en· W3095491069 on OpenAlexaff
Alison Cheung, Dan Wang, Kela Liu, Fiona Ginty, Sharon Nofech‐Mozes, Jane Bayani, John M.S. Bartlett, Anne L. Martel, Martin J. Yaffe

Bibliographic record

VenueCancer Research · 2020
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicAdvanced Biosensing Techniques and Applications
Canadian institutionsOntario Institute for Cancer ResearchHealth Sciences CentreSunnybrook Health Science Centre
Fundersnot available
KeywordsEstrogen receptorTissue microarrayBreast cancerImmunofluorescenceBiologyProgesterone receptorImmunohistochemistryCancer researchTumor microenvironmentPathologyMultiplexCancerAntibodyMedicineImmunologyTumor cellsBioinformatics

Abstract

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Abstract The extent of intra-tumoral heterogeneity - the variation in the composition of cells in a given tumor and those in the local tumor microenvironment - could have potential impact on diagnosis, treatment planning and subsequent response to treatment. To evaluate the extent of cancer cellular heterogeneity, we conducted quantitative protein marker multiplex imaging to study the variations in protein marker expression patterns on individual cells and spatial localizations. A multiplexed immunofluorescence imaging platform (MxIF, Cell DIVE) was used to measure the cellular expression of Estrogen Receptor (ER), Progesterone Receptor (PR), Epidermal Growth Factor Receptor 2 (HER2), Ki67, p53, p21WAF1 and p16INK4A in cancer epithelium. Analysis was conducted on a tissue microarray (TMA) representing subtypes classified as Luminal A-like, Luminal B-like (HER2-negative), Luminal B-like (HER2-positive), HER2-positive (non-luminal) or Triple-negative based on tumor grade and immune activity according to the St. Gallen surrogate classification. Of the 101 cores from 59 cases studied, high levels of heterogeneity were observed in ER and PR expression among the hormonal receptor-positive tumors. As expected Luminal A-like cancers exhibited higher proportions of individual cells co-expressing ER and PR, while cells in Luminal B-like, HER2-negative cancers showed ER expression only. Luminal B-like, HER2-positive cores were composed of cells with strong HER2 staining, and some cells co-expressing PR and HER2. Single cells with strong ER and HER2 labelling were rarely observed. Spatial visualizations illustrated that cells with similar expression signatures tend to be clustered together. Among cases which showed p53 overexpression with immunohistochemistry, the overall MxIF-measured p53 level was highest in TNBC compared to HER2+ and Luminal B-like cases. TNBC exhibited the highest proliferative fraction and most incidence of abnormal p53 and p16. We did not observe an association of p21 expression to P53 or P16 patterns, yet a slightly higher proportion of Luminal B-like cancers showed increased P21 levels compared to the other subtypes. Our study demonstrated the application of protein marker multiplexing and quantitative image analysis in measuring heterogeneity of protein co-expression signatures within breast cancer subtypes. Our next step is to apply the methods developed here to study a cohort where molecular profiling and radiomics were conducted (Bayani et al.) to reveal the extent of heterogeneity of breast cancer with a multi-omics approach. Citation Format: Alison M. Cheung, Dan Wang, Kela Liu, Fiona Ginty, Sharon Nofech-Mozes, Jane Bayani, John M.S. Bartlett, Anne Martel, Martin J. Yaffe. Protein marker heterogeneity in breast cancer subtypes measured using immunofluorescence protein multiplexing and quantitative, single cell image analysis [abstract]. In: Proceedings of the AACR Virtual Special Conference on Tumor Heterogeneity: From Single Cells to Clinical Impact; 2020 Sep 17-18. Philadelphia (PA): AACR; Cancer Res 2020;80(21 Suppl):Abstract nr PO-084.

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How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.010
Threshold uncertainty score0.653

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.001
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.082
GPT teacher head0.388
Teacher spread0.306 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations0
Published2020
Admission routes1
Has abstractyes

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