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kendomaniac authored Oct 18, 2023
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Expand Up @@ -85,22 +85,20 @@ We have done a 10XGenomics scRNAseq experiment including the following elements:
- DV90, 2998 sequenced cells (ERBB2 p.V842I, increases kinase activity, PMID: [*Boese et al. (2013)*](https://pubmed.ncbi.nlm.nih.gov/23220880/)
- HCC78, 2748 sequenced cells (SLC34A2-ROS1 Fusion, ROS1 inhibitors have antiproliferative effect PMID: [*Davies et al. (2012)*](https://pubmed.ncbi.nlm.nih.gov/22919003/)
- CCL.185.IG, 6354 sequenced cells [*EML4-ALK Fusion-A549 Isogenic Cell*](https://www.atcc.org/products/ccl-185ig)
- White cells from donor buffy coat (PBMC 478 sequenced cells)

<img border="0" src="../../images/drivers_minimal_relations.jpg" width="500px">

The above figure described the driver genes associated to each cell line. Only a minimal part of the connections has been shown to easy readability of the image. Full list of the interactions depicted by [*IPA*](https://digitalinsights.qiagen.com/products-overview/discovery-insights-portfolio/analysis-and-visualization/qiagen-ipa/?utm_source=QDI_GA_IPA&cmpid=QDI_GA_IPA&gclid=CjwKCAjwpuajBhBpEiwA_Ztfha6eGDCSYCgWJdU_WLaESr4az3OOXllkow2o0Xmfxz8OhFo7Z2cpxhoC4okQAvD_BwE) are available at figshare [10.6084/m9.figshare.23284748 (This doi is not active, yet)]. All driver genes have been observed in resistence occuring upon treatment with [*Isomertinib*](https://en.wikipedia.org/wiki/Osimertinib) of EGFR mutated lung cancers [[*Gomatou et al. (2023)*](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9913144/)].

PBMC will be isolated from healthy donor and snap frozen. Before single cell generation, PBMC will be marked with Biolegend TotalSeq™-B human universal cocktail v1.0.
10XGenomics analysis will be performed using CellPlex 10XGenomics technology, allowing samples labelling.

The experiment will be done using CellPlex technology from 10XGenomics allowing multiplexing samples into a single channel and therefore removing unwanted batch effects.
The experiment was done using CellPlex technology from 10XGenomics allowing multiplexing samples into a single channel and therefore removing unwanted batch effects.

Immunological cell types from PBMC will be annotated thanks to the help of an experienced immunologist.

The count tables from the entire BE1 experiment will be made available through an R Shiny app, allowing users to construct datasets encompassing different cell lines at varying ratios.
The count tables from the entire BE1 experiment are available through an [*R Shiny app*](http://130.192.212.153:3838/), allowing users to construct datasets encompassing different cell lines at varying ratios. An R package providing the same functionalites of the R Shiny app is available at [*github.com/kendomaniac/BE1*](https://github.com/kendomaniac/BE1).

**Actual state of the project:** Fully sequenced (PBMC library few cells, antibody labelling failed), raw data deposited to GEO: GSE243665 (Raw data under embargo till the pubblication of the dataset).
**Actual state of the project:** Fully sequenced, raw data deposited to GEO: GSE243665 (Raw data under embargo till the pubblication of the dataset).

<img border="0" src="../../images/be1.png" width="700px">

The above figure show the sequencing statistics of the 7 cell lines.

**Processed data are available at:** 10.6084/m9.figshare.23939481

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