Mutter-de Wit, Sanne

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Single tube liquid biopsy for NSCLC, in Advances in Circulating Tumour Cells

University of Twente Research Information

EpCAM+DAPI+CK+CD45–Round> 4µm in sizeDAPI-CK overlay >50%one tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnostic biomarkers epcam negativecirculating tumor cells liquid biopsypredictive biomarkers tumor derivedextracellular vesicles liquid biopsycirculating tumor dna prognosticbiomarker liquid biopsy liquid biopsyone tube liquid biopsy cancer patientscirculating tumor cells mutationsliquid biopsy ctc treatment epcampositive circulating tumor cells dnadiagnosticLiquid biopsy Presence in patientsGenetic analysis Overall survivalSingle tube liquid biopsy for NSCLCSanne de Wit1, Elisabetta Rossi2,3, Ellen Heitzer4, Menno Tamminga5, Mariangela Manicone3, Joost Swennenhuis1, Riccardo Vidotto3, Antonella Facchinetti2,3, Leonie Zeune1,6, Ed Schuuring5, Rita Zamarchi3, Jeroen Hiltermann3, Michael Speicher4, Leon Terstappen1, Harry Groen5CONCLUSION Presence of EpCAM+ CTC, tdEV and ctDNA are associated with poor overall survival in NSCLC and are significant predictors for OS, whereas EpCAM–/low CTC are not. What treatment relevant information can be extracted from these predictive biomarkers?1 Department of Medical Cell BioPhysics, University of Twente, The Netherlands  2  DISCOG, University of Padova, Italy  3 IOV-IRCCS, Italy  4 Institute of Human Genetics, Medical University of Graz, Austria  5 Department of Pulmonology, University of  Groningen, University Medical Centre Groningen, The Netherlands  6 Department of Applied Mathematics, University of Twente, the NetherlandsBiomarker present in 97 patients≥ 2 EpCAM+ CTC 21% All 4 biomarkers positive 02%≥ 2 EpCAM– CTC 15% 3 biomarkers positive 09%≥ 19 tdEV 27% 2 biomarkers positive 13%≥ 10% ctDNA 19% 1 biomarker positive 19%STUDY 97 Non-small cell lung cancer (NSCLC) patients before treatment were analyzed with:MedianMeanMinMax040186010692803812.86.60.260.4All from a single tube of bloodctDNA In CellSave or EDTA plasma samples the presence of >10% mutant alleles in ctDNA was determined with the mFAST-SeqS method. Isolation and sequencing of plasma for circulating tumor DNA (ctDNA)MUTATIONS IN PLASMA 14 patients were sequenced for mutations detected in primary tumor: All patients had <10% mutant alleles1 patient with EpCAM+ CTC 5 patients with EpCAM– CTC 1 patient with EpCAM+ and EpCAM– CTC 7 patients without any CTC The mutation was found in all patients with CTC In 3 patients without CTC the mutation was found as wellGenetic confirmation of tumorous origin of EpCAM–CTC on the microsieve are ongoing by detecting mutations also found in the primary tumor.contact: s.dewit@utwente.nl          www.utwente.nl/tnw/mcbp/          ACTC 2017, Rhodes, GreeceCOUNTING The presence of biomarkers above the threshold (green bar) was determined for each patient. 43% of the patients have one or more biomarkers present in their blood.EpCAM–/lowDAPI+CK+CD45–DAPI-CK overlay EpCAM+DAPI–CK+CD45–Slightly roundSurface < 150 µm2Perimeter > 4 µmDefinition CellSearch for EpCAM positive CTCMicrosieve filtration for EpCAM negative or low CTCDefinitionDefinition ACCEPT software on CellSearch cartridges for tumor derived extra-cellular vesicles (tdEV)MUTATION DETECTION IN EpCAM– CTC

Single tube liquid biopsy for NSCLC, in Advances in Circulating Tumour Cells

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