As a response to the Call for projects 2018 : Innovative technologies:


Multi-parametric analysis of cellular states at the single-cell level is enabled by cytometry-based technologies, single-cell sequencing and microscopy-based technologies. Mass cytometry enables analysis of millions of cells, but is limited by the number of parameters that can be simultaneously analyzed, usually not more than 50. Single-cell sequencing enables genome-wide analysis, although the number of cells is limited to a few hundreds, and although it is costly with data analysis that can introduce biases. Microscopy provides important spatial information but is limited by the number of parameters that can be simultaneously visualized in single cells (3 to 8, routinely).

Here, through a unique single-cell barcoding technology, hundreds of parameters in millions of single cells will be analyzed, enabling a sequencer to provide cytometry-like data. The same technology can be applied for imaging readouts, allowing medium-to-high throughput analysis of proteins and RNAs. With such technology, we aim at getting deep insights in T-lymphocytes “exhaustion”, which is a hallmark of ineffective cancer immunotherapy. Understanding the molecular mechanisms that drive such dysfunctional states is pivotal to the effectiveness of clinical immune modulation. Publicly- available data and our preliminary studies suggest that, in the way of becoming dysfunctional, T- lymphocytes accumulate DNA-damage. Within the pipeline, we will also develop a method for DNA- damage detection in single-cells.

Overall, the goal of this collaborative project is to melt the technological knowledge of Dr. Bava with the deep cancer-immunology background of Dr. Amigorena, and to advance our knowledge in the field of cancer immunology, towards more efficient immunotherapies.

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