Identification and quantification of malignant cells
(CSFTCs) in cerebrospinal fluid :
GC Faure, Q Tu (Nancytomique- Nancy...
Plan
• Leptomeningeal Metastases (LM)
• The biological fluid CSF
• Diagnostic challenges Gold Standard
• Method
• Results
...
LM or meningeal carcinomatosis
Epidemiology... Prognosis
 3 to 5% of cancer patients, incidence up to 9.6% (J Clin
Oncol ...
Cerebrospinal fluid: CSF
• Volume 150mL
• Production #500mL per day, (3.7x)
• Choroid plexuses
• Lumbar puncture Berlin
– ...
LM: Diagnostic challenge
Gold standard
Dux et al, J Neurol Sci, 1994; 121; 74-78
 CSF volume
– 3.5mL: 68% positivity
– 10...
Methods
• CellSave® tubes allowing multicentric
studies
• CellSearch® Veridex
– Cell Tracks AutoPrep
– Analyzer II... Mult...
LM and « CSFTCs » in Breast
Cancer (BMC Clinical Pathology 2012)
• Detection
• Quantification: 1 to 103-4
• Characterizati...
Sequential analysis of CSFTCs
confirms repetability of numerations
with two subgroups (high > 700/mL vs low)
BMC Clin Path...
Melanoma CSFMCs
Medical Oncology 2013;
• CMC kit (J&J, VERIDEX)
– CD146 capture, MEL14 , CD45
• Four patients 9 points
• G...
LUNG Cancer
CSFTCs + CSFTMs
Cytomorphology: cell galleries
allow to differentiate SCLC, NSCLC
(adenocarcinoma, squamous ca...
CSFTCs: a new frontier!
• Tumoral (epithelial) cells can be detected
and quantified in CSF with the CellSearch®
technology...
Clinical, Basic, Therapeutic
openings
• Evaluation of systemic and intrathecal therapies in
clinical settings
• Comparison...
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Cytology Paris 2013: presentation selected for a cytopathologist junior prize

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Identification and quantification of malignant cells (CSFTCs) in cerebrospinal fluid by GC Faure, Q Tu, E Le Rhun

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Cytology Paris 2013: presentation selected for a cytopathologist junior prize

  1. 1. Identification and quantification of malignant cells (CSFTCs) in cerebrospinal fluid : GC Faure, Q Tu (Nancytomique- Nancy), E Le Rhun (Lille) We declare not having a conflict of interest Diagnosis of leptomeningeal metastases (LM) in patients with solid tumors (breast, lung, prostate...) and melanomas remains difficult. Usual diagnostic methods of cytomorphological assessment of cerebro-spinal fluid (CSF) and gadolinium enhanced MRI lack both specificity and sensitivity.
  2. 2. Plan • Leptomeningeal Metastases (LM) • The biological fluid CSF • Diagnostic challenges Gold Standard • Method • Results • Openings
  3. 3. LM or meningeal carcinomatosis Epidemiology... Prognosis  3 to 5% of cancer patients, incidence up to 9.6% (J Clin Oncol 2004;22:2865)  Up to 19% of autopsied patients with cancer and neurological symtoms (Glass, 1979) – Breast cancer (5%), lung (11%), melanoma (20%)  Increasing incidence – Better survival of cancer patients + New molecules for systemic disease have bad meningeal diffusion Kodack DP et al. PNAS 2012, 109, E3119  Very Bad prognosis (4 weeks to 6 months) and bad quality of life – But promise of new intrathecal drugs (MTX, trastuzumab...) and trials (Chamberlain)
  4. 4. Cerebrospinal fluid: CSF • Volume 150mL • Production #500mL per day, (3.7x) • Choroid plexuses • Lumbar puncture Berlin – Heinrich Ireneus Quincke – Berl klin Wochenschr 1891;28:929 +965
  5. 5. LM: Diagnostic challenge Gold standard Dux et al, J Neurol Sci, 1994; 121; 74-78  CSF volume – 3.5mL: 68% positivity – 10.5mL: 97% positivity  Time interval between sampling and analysis  Cell viability 30 mns 50%; 60 mns 20%; 90 mns 10%  Good sensitivity requires  First LP 40%  Second LP 80%, Third LP to reach 90-95%  No reliable quantification – Response at 50% threshold
  6. 6. Methods • CellSave® tubes allowing multicentric studies • CellSearch® Veridex – Cell Tracks AutoPrep – Analyzer II... Multispectral imaging • CellTracks® CTCs and CMC kits – EpCAM enrichment, KT label, CD45 • Validation on image galeries CellTracks® AutoPrep® System CellTracks® Analyzer II Cancer type Breast Lung Melanoma Tumorcells/5mLLCR 0,1 1 10 100 1000 10000 100000
  7. 7. LM and « CSFTCs » in Breast Cancer (BMC Clinical Pathology 2012) • Detection • Quantification: 1 to 103-4 • Characterization – Her2-Neu Specificity: no contaminating ependymal cells Sensitivity: detection and quantification in all established LM patients studied compared to cytology High homogeneity (and reproducibility) of images High purity compared to blood samples
  8. 8. Sequential analysis of CSFTCs confirms repetability of numerations with two subgroups (high > 700/mL vs low) BMC Clin Pathol 2012 Sample number 1st 2nd 3rd 4th 5th Tumoralcells/5mLLCR 0,1 1 10 100 1000 10000 100000 DM CTC WA CTC CJ CTC DMB CTC VT CTC PV CTC BE CTC CS CTC HE CMC ST CMC DC CTC
  9. 9. Melanoma CSFMCs Medical Oncology 2013; • CMC kit (J&J, VERIDEX) – CD146 capture, MEL14 , CD45 • Four patients 9 points • Good reproducibility during follow-up • Cell morphology of melanoma cells in CSF far better than in blood
  10. 10. LUNG Cancer CSFTCs + CSFTMs Cytomorphology: cell galleries allow to differentiate SCLC, NSCLC (adenocarcinoma, squamous carcinoma) and detect apoptosis, autophagy... • Numerous CTMs in some NSCLC patients up to 80%, in sequential study
  11. 11. CSFTCs: a new frontier! • Tumoral (epithelial) cells can be detected and quantified in CSF with the CellSearch® technology (CSFTCs) • Their numbers can be sequentially followed- up in breast, lung and other cancers – allowing to evaluate the efficacy of treatments (intrathecal and/or systemic) • Tumoral cell population in CSF might be different from blood CTCs, allowing further studies of metastatic properties • CSFMCs can also be detected and quantified in CSF
  12. 12. Clinical, Basic, Therapeutic openings • Evaluation of systemic and intrathecal therapies in clinical settings • Comparison with other CTCs tecnologies (Wuhan CTC chip, … ScreenCell, ImageStreamX AMNIS flow, EPIC on slide) • Early Clinical detection of brain and meningeal metas! • Characteristics of cells going through the BBB (HER-2, EGF-R, etc..)... understanding metastasis • Enrichment/isolation for further cell and molecular biology studies. ... (Parrot project: lung and prostate) • New therapy targets?

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