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Edited training material "Automated Cell Annotation" upto slide 10 #5604
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### What is Cell Annotation? | ||||||
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- Categorising cells into cell type categories based on transcriptomic data | ||||||
- Classify cells in your data into different cell types based on gene expression data | ||||||
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- Can be done manually or automatically | ||||||
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- Is possible due to single-cell sequencing technology | ||||||
- Single-cell sequencing technology provides higher resolution than bulk RNA-seq | ||||||
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![Basic pipeline for automated cell annotation](../../images/scrna-cell-annotation/cell-annotation.png) | ||||||
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--- | ||||||
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### Why is it Important? | ||||||
### Why is Cell Annotation Important? | ||||||
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- Able to process and analyse single cell data much faster than manual analysis | ||||||
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- To understand the composition of cell types in samples (cellular heterogeneity) | ||||||
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- To compare changes in cell populations or states across different condition and phenotypes | ||||||
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Suggested change
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- To perform differential expression within each cell type to avoid signal dilution from mixed cell type population | ||||||
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- To identify novel cell states and study some cell population further | ||||||
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??? | ||||||
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The power of single-cell RNA-seq lies in its ability to capture the transcriptome at single-cell resolution. | ||||||
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However, a significant challenge is accurately classifying cells into distinct types before beginning downstream analysis. | ||||||
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Once cells are annotated, we can examine the composition of cell types within each sample and compare them across conditions. | ||||||
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This precise classification enables differential expression analysis within specific cell types, a key objective of single-cell experiments. | ||||||
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--- | ||||||
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### Why Automate Cell Annotation? | ||||||
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- Each single-cell experiment can generate data for thousands of cells | ||||||
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- Manual annotation is time consuming and requires domain expertise | ||||||
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- Can produce results more consistently allowing for reproducibility of results | ||||||
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- Cell annotation uses sc-RNA seq data | ||||||
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- Gene expressions are stored in a gene expression matrix (X) | ||||||
- Gene expressions are stored in a gene expression matrix (X) <br><br> | ||||||
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Common data types: | ||||||
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--- | ||||||
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### Challenges | ||||||
### Challenges in Automated Cell Type Annotation | ||||||
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- Noise due to amplification techniques, varying sequencing depths, and sequencing errors | ||||||
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- Data is often noisy due to amplification techniques, varying sequencing depths, and errors in the reads | ||||||
- Difference in QC steps for reference and query dataset may introduce a bias | ||||||
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- Cell type definitions are inherently subjective and may be suboptimal | ||||||
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- Lack of a suitable reference panel for the query dataset can result in inaccurate classification, especially when dealing with unknown cell states | ||||||
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- Dealing with unknown cell types due to undiscovered classifications | ||||||
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??? | ||||||
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There are some challenges that need to be faced in order to perform automated cell annotation: | ||||||
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### Manual Cell Annotation | ||||||
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- Generate a cluster map and manually annotate each clusters | ||||||
- Requires known marker genes for cell types of interest | ||||||
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- Generate the UMAP/tSNE (see below) to visualize the expression values | ||||||
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- Requires the researchers to find known marker genes in the data | ||||||
- If you have clustered the cells, you can use dotplots or violin plots to measure the average expression of these genes | ||||||
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![Various cell cluster diagrams showing the expression values of various marker genes](../../images/scrna-cell-annotation/manual-annotation.png) | ||||||
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.pull-left[ | ||||||
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- The process of identifying unique gene expressions that can be used for identifying cell types | ||||||
- The process of identifying genes that are uniquely expressed in certain cell types | ||||||
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- Genes are selected based on how differently they are expressed across different cell types | ||||||
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I feel like this point is missing something to explicitly link it to the cell annotation - maybe combine it with the start of the old version like 'Is possible because single-cell seq tech provides higher res...'?