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06 explore infercnv for Wilms tumor -06 #828

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991ff2a
Start exploring infercnv results
maud-p Oct 16, 2024
f8fff21
update notebook README.md file
maud-p Oct 16, 2024
07973d3
Update README.md
maud-p Oct 16, 2024
ff057bb
changes to PR#828
maud-p Oct 18, 2024
f06b6a9
PR#828 Add corrected html notebooks
maud-p Oct 19, 2024
617d822
corrections PR#828
maud-p Oct 19, 2024
84286eb
improve PR#828 and try inter-patient normal reference
maud-p Oct 21, 2024
52cf2aa
add ggdist dependencie
maud-p Oct 28, 2024
eced971
Merge pull request #9 from maud-p/origin/06_explore_infercnv
maud-p Oct 28, 2024
e4cff23
few changes to PR828
maud-p Oct 28, 2024
2e6fd9a
few changes to PR828
maud-p Oct 28, 2024
d25e463
Merge pull request #10 from maud-p/origin/06_explore_infercnv
maud-p Oct 28, 2024
176d5ff
update notebook
maud-p Oct 28, 2024
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Merge pull request #11 from maud-p/origin/06_explore_infercnv
maud-p Oct 28, 2024
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add comment on 06_infercnv.R on saved output
maud-p Oct 29, 2024
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Merge pull request #12 from maud-p/origin/06_explore_infercnv
maud-p Oct 29, 2024
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Update the main README.md file
maud-p Oct 29, 2024
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Merge branch 'main' into 06_explore_infercnv
sjspielman Oct 29, 2024
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Update analyses/cell-type-wilms-tumor-06/scripts/06_infercnv.R
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Update analyses/cell-type-wilms-tumor-06/scripts/06_infercnv.R
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18 changes: 12 additions & 6 deletions analyses/cell-type-wilms-tumor-06/notebook/README.md
Original file line number Diff line number Diff line change
Expand Up @@ -15,25 +15,25 @@ As part of the `00b_characterize_fetal_kidney_reference_Stewart.Rmd` notebook te
Each of the sample of the Wilms tumor dataset SCPCP000006 as been pre-processed and characterized as the following.
Reports for each of the steps are found in the notebook/{sample_id} directory:

- `01_seurat_processing_{sample-id}.html` is the output of the [`01_seurat-processing.Rmd`](../notebook_template/01_seurat-processing.Rmd) notebook template.
- [x] `01_seurat_processing_{sample-id}.html` is the output of the [`01_seurat-processing.Rmd`](../notebook_template/01_seurat-processing.Rmd) notebook template.
In brief, the `_processed.rds` `sce object` is converted to `Seurat` and normalized using `SCTransform`.
Dimensionality reduction (`RunPCA` and `RunUMAP`) and clustering (`FindNeighbors` and `FindClusters`) are performed before saving the `Seurat` object.

- `02a_fetal_full_label-transfer_{sample-id}.html` is the output of the [`02a_label-transfer_fetal_full_reference_Cao.Rmd`](../notebook_template/02a_label-transfer_fetal_full_reference_Cao.Rmd) notebook template.
- [x] `02a_fetal_full_label-transfer_{sample-id}.html` is the output of the [`02a_label-transfer_fetal_full_reference_Cao.Rmd`](../notebook_template/02a_label-transfer_fetal_full_reference_Cao.Rmd) notebook template.
In brief, we used `Azimuth` to transfer labels from the `Azimuth` fetal full reference (Cao et al.)

- `02b_fetal_kidney_label-transfer_{sample-id}.html` is the output of the [`02b_label-transfer_fetal_kidney_reference_Stewart.Rmd`](../notebook_template/02b_label-transfer_fetal_kidney_reference_Stewart.Rmd) notebook template.
- [x] `02b_fetal_kidney_label-transfer_{sample-id}.html` is the output of the [`02b_label-transfer_fetal_kidney_reference_Stewart.Rmd`](../notebook_template/02b_label-transfer_fetal_kidney_reference_Stewart.Rmd) notebook template.
In brief, we used `Azimuth` to transfer labels from the fetal kidney reference (Stewart et al.)

- `03_clustering_exploration_{sample-id}.html` is the output of the [`03_clustering_exploration.Rmd`](../notebook_template/03_clustering_exploration.Rmd) notebook template.
- [x] `03_clustering_exploration_{sample-id}.html` is the output of the [`03_clustering_exploration.Rmd`](../notebook_template/03_clustering_exploration.Rmd) notebook template.
In brief, we explore the clustering results, we look into some marker genes, pathways enrichment and label transfer.


## Global analysis

The next step in analysis is to identify tumor vs. normal cells.

- `04_annotation_Across_Samples_exploration.html` is the output of the [`04_annotation_Across_Samples_exploration.Rmd`](../notebook/04_annotation_Across_Samples_exploration.Rmd) notebook.
- [x] `04_annotation_Across_Samples_exploration.html` is the output of the [`04_annotation_Across_Samples_exploration.Rmd`](../notebook/04_annotation_Across_Samples_exploration.Rmd) notebook.
In brief, we explored the label transfer results across all samples in the Wilms tumor dataset SCPCP000006 in order to identify a few samples that we can begin next analysis steps with.

## Exploratory analysis
Expand All @@ -45,8 +45,14 @@ We selected in [`04_annotation_Across_Samples_exploration.Rmd`](../notebook/04_a
- sample SCPCS000205 has > 89 % of cells predicted as kidney and 92 + 76 endothelium and immune cells.
- sample SCPCS0000208 has > 95 % of cells predicted as kidney and 18 + 35 endothelium and immune cells.

We wanted to test `copykat` results obtained with or without normal cells as reference, using either an euclidean or statistical (spearman) method for CNV heatmap clustering.
- [x] `05_copykat_exploration_{sample_id}.html` is the output of the [`05_copykat_exploration.Rmd`](../notebook_template/05_copykat_exploration.Rmd) notebook template.

In brief, we wanted to test `copykat` results obtained with or without normal cells as reference, using either an euclidean or statistical (spearman) method for CNV heatmap clustering.
This impact the final decision made by `copykat` for each cell to be either aneuploid or diploid, and it is thus crucial to explore the results using the different methods.
For each of the selected samples, we explore the results in the template `notebook` [`05_copykat_exploration.Rmd`](../notebook_template/05_copykat_exploration.Rmd), which creates a notebook `05_cnv_copykat_exploration_{sample_id}.html` for each sample.
These `notebooks` are inspired by the plots written for the Ewing Sarcoma analysis in [`03-copykat.Rmd`](https://github.com/AlexsLemonade/OpenScPCA-analysis/blob/main/analyses/cell-type-ewings/exploratory_analysis/03-copykat.Rmd).

- [x] `06_infercnv_exploration_{sample_id}.html` is the output of the [`06_infercnv_exploration.Rmd`](../notebook_template/06_infercnv_exploration.Rmd) notebook template.

In brief, we wanted to test `infercnv` results obtained with or without endothelium and/or immune cells as reference.
We also explore the potential of using a HMM model to assign CNV scores for each cells and discriminate normal from cancer cells.

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