Poster Presentation 35th Lorne Cancer Conference 2023

Time course analysis of pseudo-bulk samples from single cell RNA sequencing data (#123)

Jinming Cheng 1 2 , Yunshun Chen 1 2 3 , Gordon Smyth 1 4
  1. Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia
  2. Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia
  3. ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia
  4. School of Mathematics and Statistics, The University of Melbourne, Parkville, VIC , Australia

Single cell RNA sequencing technologies have been rapidly developed in recent years. The 10x droplet-based single cell RNA sequencing technology makes it possible to profile gene expression of tens of thousands of cells per sample. Standard analysis of single cell RNA sequencing data usually includes quality control, normalization, dimension reduction, cell clustering and differential expression analysis. Removing the potential doublets is also recommended in the the standard analysis. Multiple samples at different stages can be integrated together, and the downstream trajectory analysis can be performed to study the cell development process. Here, we further extend the downstream analysis to time course analysis taking advantage of the pseudotime inferred from trajectory analysis. In this workflow, we use single cell RNA sequencing data of mouse mammary gland epithelium at five different stages to demonstrate the standard analysis and integration analysis with Seurat, the doublet prediction with scDblFinder, the ternary plot analysis using signature genes, the trajectory analysis with monocle3, and time course analysis using pseudo-bulk data with edgeR.

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  2. Pal, B. et al. Single cell transcriptome atlas of mouse mammary epithelial cells across development. Breast Cancer Res 23, 69 (2021).
  3. Hao, Y. et al. Integrated analysis of multimodal single-cell data. Cell 184, 3573-3587 e3529 (2021).
  4. Cao, J. et al. The single-cell transcriptional landscape of mammalian organogenesis. Nature 566, 496-502 (2019).