Isolation of rare circulating tumor cells in cancer patients by microchip technology Nagrath & Sequist et al. (2007) Journal Presentation Brigitte Morales.

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Presentation transcript:

Isolation of rare circulating tumor cells in cancer patients by microchip technology Nagrath & Sequist et al. (2007) Journal Presentation Brigitte Morales & Leanna Morinishi December 8 th, 2011 Nagrath, S., Sequist, L. et al. Nature (2007): Print.

Circulating Tumor Cells Viable cancer cells detached from the primary tumor and circulating through bloodstream May “seed” additional tumors (metastasis) Very rare (1 in 10 9 ) b.examiner.com/sites/default/files/styles/large/hash/df/20/df20e5 2fb60caa56c75fbf5463efb422.jpg

Previous research CTC studies potential in cancer research o Circumvent invasive biopsies o Further cancer marker studies o Study metastasis Previous methods of CTC isolation o Requires pre-processing of blood o Identification after fixed cells o Poor yield and purity

Microchip for high-quality isolation

Design considerations Micro-post placement maximizes probability of contact Optimize flow velocity Minimize shear force

CTC capture Cancer cells of interest overexpress EpCAM Haematologic cells do not Coated microposts with anti-EpCAM antibodies capture CTCs Non-specifically bound cells are washed out after initial flow es/Nov08SeriesImage.jpg

Controls Non-small-cell lung cancer cells spiked into buffer o Capture efficiency: 65% 3 cell lines with varying antigen expression o Capture efficiency: >65% Density of antigens/cell doesn’t affect the number of cells captured NSCLC cells spiked into whole and lysed blood from healthy donors o CTC-chip doesn’t require blood sample pre-processing Examined blood samples from 20 healthy individuals

Visual confirmation CTCs - rhodamine-conjugated anti-cytokeratin Haematologic cells – fluorescein-conjugated anti- CD45 antibodies Both – DAPI staining for DNA

Longitudinal tumor study

Major Conclusions Efficient - CTC isolation, high purity and yield Simple - Analyzes whole blood, in one-step Gentle - Isolates viable cells Powerful - Useful for early diagnosis and longitudinal monitoring of cancer Future Potential for expansion into other disease studies

Advances and Significance Improvements o Recovering the live cells o Using this device for capture of other rare cells Post-2007, cited in >400 articles o Span topics from metastasis to lab-on-a-chip As of Oct., antibody-based capture still the purest yield technique

Questions?

Nagrath, S., Sequist, L., Maheswaran, S., Bell, D., Irimia, D., Ulkus, L., Smith, M., Kwak, E., Digumarthy, S., Muzikansky, A., Ryan, P., Balis, U., Tompkins, R., Haber, D., Toner, M. “Isolation of Rare Circulating Tumour Cells in Cancer Patients by Microchip Technology.” Nature (2007): Print.