Blanc Inter SIMI 9 - Blanc international - Sciences de l'information, de la matière et de l'ingénierie : Sciences de l'ingénierie, matériaux, procédés, énergie

Developing new microfluidic chambers for cancer cell screening – ONCO-SCREEN

Submission summary

The research program presented here is jointly conducted by Singaporean and French partners with strong expertise in biophysics, cancer biology, ultra-sensitive analytical chemistry and microfluidics who wish to implement an integrated microsystem designed to simultaneously quantitate motility and gene expression of live cancer cells challenged by different chemical environments.
Cancer cell migration will be induced and directed by a concentration gradient of chemoattractant generated by a microfluidic system and cells will be separated and collected in individual chambers according to their velocity. We have demonstrated that cells migrate as expected in the microfluidic device and we now wish trough this franco-singaporean partnership to integrate in the microfluidic device an ultrasensitive biosensor arrays for on site and on chip molecular analyses of gene expression. Our goal is to provide the community with a scientific tool that will allow thorough characterisation of cancer cells based on the quantitative characterisation of their activity (migration, velocity) and molecular signature (gene expression) and to examine how these characteristics change with the nature of their environment.
A microfluidic device that diffuses nanoliter volumes of a unidirectional chemoattractant molecule (Epidermal Growth Factor or stromal-derived factor-1) into a microchamber will be loaded with three human cancer cell lines (MCF-7, HBL-100, MDA-MB-231) known for their strikingly different migratory properties. Quantitative analysis of cell migration, velocity and polarisation will be carried out by real-time microscopy and computer-assisted. Cancer cells will be collected (either by using trypsin or a hydrodynamic protocol) and processed for routine molecular analysis using quantitative reverse-transcriptase-polymerase chain reaction (qRT-PCR) of selected markers including epithelial differentiation markers (E-cadherin, beta-catenin) and mesenchymal cell markers (Vimentin, twist). Next, a protocol will be developed to ensure on-chip detection of specific messengers ribonucleic acids (mRNA) that are only expressed in MCF-7 (oestrogen receptor), HBL-100 (activin ßa) and MDA-MB-231 (vimentin) cells. Following their migration in the chemotaxis chamber, each cancer cell population will be collected in individual chambers and exposed to an electrical field to induce total cell membrane rupture. Released cytoplasmic mRNA will be pushed towards the detection area to carry out molecular analyses using a nanostructured electrical sensor array. Following hybridization between the target mRNA and the immobilized complementary probe, a bridging mechanism will enable the production of an electrical conductivity. Measured current will be proportionally correlated to the amount of stable hybrids formed and will permit to correlate a gene signature with the phenotype of each cancer cell line and to examine the effect of changing chemical environments on the phenotype and genotypic expression.
This project will allow the production of an ultrasensitive electrical sensor array-based system in the shortest possible time, providing a powerful tool for vast molecular analyses in biological research. The knowledge derived from this project will have tremendous potential benefits through the development of a new lab-on-chip screening device for cancer cells and will further accelerate the transfer of microfluidic and nano-gap sensor technologies to human cancer diagnosis and prediction

Project coordination

Rosaria FERRIGNO (UNIVERSITE CLAUDE BERNARD - LYON I) – rosaria.ferrigno@univ-lyon1.fr

The author of this summary is the project coordinator, who is responsible for the content of this summary. The ANR declines any responsibility as for its contents.

Partner

INL - UCBL UNIVERSITE CLAUDE BERNARD - LYON I
CGMC - UCBL UNIVERSITE CLAUDE BERNARD - LYON I

Help of the ANR 236,087 euros
Beginning and duration of the scientific project: - 36 Months

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