Cachectic multiome human
  • Interactive overview
  1. Cachectic Multiome Human
  • Cachectic Multiome Human

On this page

  • Study Map
  • Quality Control
  • Annotation
  • Differential Expression
  • DEVIL Cell-Level DE
  • GSEA And Pathways
  • Cell-Cell Communication
  • Figure Gallery
  • Legacy Presentations
  • Repository Map
  • Discussion Questions
  • Meeting Notes

Cachectic Multiome Human

Interactive discussion document

Author

Sophie Grace Parolin

Published

May 15, 2026

Core aim

Use this page as the live presentation surface for the cachexia multiome project: start with the main biological story, then jump into QC, annotation, DEG, GSEA, DEVIL, or CCC only when someone asks.

Status

This is the scaffold. It is ready for discussion notes and exported figures; the analysis scripts remain in the repository as the reproducible backend.

Jump to what they ask

Study map QC Annotation DEG DEVIL GSEA CCC Files Open questions

Study Map

Presentation principle

Keep each section to one answerable question, one primary visual, and one collapsible methods block. During a meeting, the document becomes a map rather than a long linear talk.

Step Module Key_files
0 Input metadata / TBI files 0_tbi_file*.R
1 Integration by sex NuovaPipeNo120/1_Int*_NewData.R
2 First annotation NuovaPipeNo120/2_Ann*_New.R
3 Refined annotation NuovaPipeNo120/3_Ann*_pt2_New.R
4 Gene filtering NuovaPipeNo120/4_FiltGenes_*_NewData.R
5 Pseudo-bulk differential expression NuovaPipeNo120/5_DEG*.R
6 Cell-level DE with devil NuovaPipeNo120/6_DEA_devil_*.R
7-8 GSEA / pathway interpretation NuovaPipeNo120/7_GSEA_F.R, 8_GSEA_*
10 Cell-cell communication NuovaPipeNo120/10_CCC_*.R
Talking path for a 5-minute overview
  1. What biological contrast defines cachexia in this cohort?
  2. Which nuclei/cell states are robust after QC and annotation?
  3. Which changes are shared versus sex-specific?
  4. Which pathways move consistently across pseudo-bulk, DEVIL, and GSEA?
  5. Which cell-cell communication signals are worth validating?

Quality Control

Question to answer live: are the nuclei/cells reliable enough to support the downstream story?

K017 before filtering

K017 after filtering

RNA QC

ATAC QC

Relevant scripts and logs
File
1_QualityControl.R
1_QualityControl_NewData.R
2_RemoveDoublets_Fem.R
2_RemoveDoublets_Fem_NewData.R
2_RemoveDoublets_Man.R
2_RemoveDoublets_Man_NewData.R
3_QC_ribosom.R

Annotation

Question to answer live: what cell types/states are present, and which annotations are stable across RNA, ATAC, and WNN views?

  • Female
  • Male
  • Marker Logic

Annotated UMAP

Marker dotplot

Cell composition by condition

Annotated UMAP

Marker dotplot

Cell composition by condition

Use this tab for the curated marker list and difficult annotation decisions.

Relevant scripts
File

Differential Expression

Question to answer live: which transcriptional changes separate CAC, PRE, and CTRL, and are they cluster-specific or global?

  • Pseudo-bulk
  • Female
  • Male
  • Myonuclei

The current local archive contains DEG results mainly as .xlsx tables; this tab keeps their locations close to the visual pathway summaries below.

Table
F/Cluster/DEA_FEM_CAC_vs_CTRL.xlsx
F/Cluster/DEA_FEM_CAC_vs_PRE.xlsx
F/Cluster/DEA_FEM_PRE_vs_CTRL.xlsx
F/Clusters_NoRibo/DEA_FEM_CAC_vs_CTRL.xlsx
F/Clusters_NoRibo/DEA_FEM_CAC_vs_PRE.xlsx
F/Clusters_NoRibo/DEA_FEM_PRE_vs_CTRL.xlsx
F/Myo/DEA_FEM_Myo_Overall_CAC_vs_CTRL.xlsx
F/Myo/DEA_FEM_Myo_Overall_CAC_vs_PRE.xlsx
F/Myo/DEA_FEM_Myo_Overall_PRE_vs_CTRL.xlsx
F/Myo_NoRibo/DEA_FEM_Myo_Overall_CAC_vs_CTRL.xlsx
F/Myo_NoRibo/DEA_FEM_Myo_Overall_CAC_vs_PRE.xlsx
F/Myo_NoRibo/DEA_FEM_Myo_Overall_PRE_vs_CTRL.xlsx
M/Clusters/DEA_MAN_CAC_vs_CTRL.xlsx
M/Clusters/DEA_MAN_CAC_vs_PRE.xlsx
M/Clusters/DEA_MAN_PRE_vs_CTRL.xlsx
M/Myo/DEA_MAN_Myo_Overall_CAC_vs_CTRL.xlsx
M/Myo/DEA_MAN_Myo_Overall_CAC_vs_PRE.xlsx
M/Myo/DEA_MAN_Myo_Overall_PRE_vs_CTRL.xlsx
M/old/cluster/DEA_MAN_CAC_vs_CTRL.xlsx
M/old/cluster/DEA_MAN_CAC_vs_PRE.xlsx
M/old/cluster/DEA_MAN_PRE_vs_CTRL.xlsx
M/old/cluster_noRibo/DEA_MAN_CAC_vs_CTRL.xlsx
M/old/cluster_noRibo/DEA_MAN_CAC_vs_PRE.xlsx
M/old/cluster_noRibo/DEA_MAN_PRE_vs_CTRL.xlsx
M/old/Myo/DEA_MAN_Myo_Overall_CAC_vs_CTRL.xlsx
M/old/Myo/DEA_MAN_Myo_Overall_CAC_vs_PRE.xlsx
M/old/Myo/DEA_MAN_Myo_Overall_PRE_vs_CTRL.xlsx
M/old/Myo_NoRibo/DEA_MAN_Myo_Overall_CAC_vs_CTRL.xlsx
M/old/Myo_NoRibo/DEA_MAN_Myo_Overall_CAC_vs_PRE.xlsx
M/old/Myo_NoRibo/DEA_MAN_Myo_Overall_PRE_vs_CTRL.xlsx

Use this tab for CAC vs CTRL, CAC vs PRE, and PRE vs CTRL in female samples.

Use this tab for CAC vs CTRL, CAC vs PRE, and PRE vs CTRL in male samples.

Use this tab for myonuclei-specific contrasts and comparisons with global/cluster-level results.

Relevant scripts and outputs
File

DEVIL Cell-Level DE

Question to answer live: do cell-level models support or refine the pseudo-bulk signals?

Table
F/Clusters/DEA_DEVIL_FEM_CAC_vs_CTRL.xlsx
F/Clusters/DEA_DEVIL_FEM_CAC_vs_PRE.xlsx
F/Clusters/DEA_DEVIL_FEM_PRE_vs_CTRL.xlsx
F/Clusters_NoRibo/DEA_DEVIL_FEM_CAC_vs_CTRL.xlsx
F/Clusters_NoRibo/DEA_DEVIL_FEM_CAC_vs_PRE.xlsx
F/Clusters_NoRibo/DEA_DEVIL_FEM_PRE_vs_CTRL.xlsx
F/Custom_Sample_Comparisons/DEA_DEVIL_FEM_CUSTOM_All_CTRL_vs_K048.xlsx
F/Custom_Sample_Comparisons/DEA_DEVIL_FEM_CUSTOM_All_CTRL_vs_PK120.xlsx
F/Myo/DEA_DEVIL_FEM_Myo_Overall_CAC_vs_CTRL.xlsx
F/Myo/DEA_DEVIL_FEM_Myo_Overall_CAC_vs_PRE.xlsx
F/Myo/DEA_DEVIL_FEM_Myo_Overall_PRE_vs_CTRL.xlsx
F/Myo_NoRibo/DEA_DEVIL_FEM_Myo_Overall_CAC_vs_CTRL.xlsx
F/Myo_NoRibo/DEA_DEVIL_FEM_Myo_Overall_CAC_vs_PRE.xlsx
F/Myo_NoRibo/DEA_DEVIL_FEM_Myo_Overall_PRE_vs_CTRL.xlsx
M/Clusters/DEA_DEVIL_MAN_CAC_vs_CTRL.xlsx
M/Clusters/DEA_DEVIL_MAN_CAC_vs_PRE.xlsx
M/Clusters/DEA_DEVIL_MAN_PRE_vs_CTRL.xlsx
M/Myo/DEA_DEVIL_MAN_Myo_Overall_CAC_vs_CTRL.xlsx
M/Myo/DEA_DEVIL_MAN_Myo_Overall_CAC_vs_PRE.xlsx
M/Myo/DEA_DEVIL_MAN_Myo_Overall_PRE_vs_CTRL.xlsx

The visual DEVIL-ranked pathway summaries are shown under GSEA and pathways, in the DEVIL-ranked GSEA tab.

Relevant scripts and logs
File

GSEA And Pathways

Question to answer live: which pathways make the gene-level changes biologically coherent?

  • Female GSEA
  • Male GSEA
  • DEVIL-ranked GSEA

Myonuclei CAC vs CTRL

Myonuclei CAC vs PRE

Myonuclei PRE vs CTRL

Myonuclei CAC vs CTRL

Myonuclei CAC vs PRE

Myonuclei PRE vs CTRL

Female myonuclei CAC vs CTRL

Male myonuclei CAC vs CTRL

Relevant scripts and logs
File

Cell-Cell Communication

Question to answer live: which ligand-receptor programs connect cachexia-associated states?

CAC Myo IIx to T cells: clusters

Sender clusters

Receiver clusters

Top ligand-receptor markers

Relevant scripts and logs
File

Figure Gallery

### annotation_f_cells_by_condition.png

![](assets/figures/annotation/annotation_f_cells_by_condition.png){fig-alt='annotation_f_cells_by_condition.png'}

### annotation_f_marker_dotplot.png

![](assets/figures/annotation/annotation_f_marker_dotplot.png){fig-alt='annotation_f_marker_dotplot.png'}

### annotation_f_umap_combined.png

![](assets/figures/annotation/annotation_f_umap_combined.png){fig-alt='annotation_f_umap_combined.png'}

### annotation_m_cells_by_condition.png

![](assets/figures/annotation/annotation_m_cells_by_condition.png){fig-alt='annotation_m_cells_by_condition.png'}

### annotation_m_marker_dotplot.png

![](assets/figures/annotation/annotation_m_marker_dotplot.png){fig-alt='annotation_m_marker_dotplot.png'}

### annotation_m_umap_combined.png

![](assets/figures/annotation/annotation_m_umap_combined.png){fig-alt='annotation_m_umap_combined.png'}

### ccc_myoIIx_to_tcells_clusters.png

![](assets/figures/ccc/ccc_myoIIx_to_tcells_clusters.png){fig-alt='ccc_myoIIx_to_tcells_clusters.png'}

### ccc_myoIIx_to_tcells_lr_dotplot.png

![](assets/figures/ccc/ccc_myoIIx_to_tcells_lr_dotplot.png){fig-alt='ccc_myoIIx_to_tcells_lr_dotplot.png'}

### ccc_myoIIx_to_tcells_receiver.png

![](assets/figures/ccc/ccc_myoIIx_to_tcells_receiver.png){fig-alt='ccc_myoIIx_to_tcells_receiver.png'}

### ccc_myoIIx_to_tcells_sender.png

![](assets/figures/ccc/ccc_myoIIx_to_tcells_sender.png){fig-alt='ccc_myoIIx_to_tcells_sender.png'}

### devil_gsea_f_myonuclei_cac_vs_ctrl.png

![](assets/figures/devil/devil_gsea_f_myonuclei_cac_vs_ctrl.png){fig-alt='devil_gsea_f_myonuclei_cac_vs_ctrl.png'}

### devil_gsea_m_myonuclei_cac_vs_ctrl.png

![](assets/figures/devil/devil_gsea_m_myonuclei_cac_vs_ctrl.png){fig-alt='devil_gsea_m_myonuclei_cac_vs_ctrl.png'}

### gsea_f_myonuclei_cac_vs_ctrl.png

![](assets/figures/gsea/gsea_f_myonuclei_cac_vs_ctrl.png){fig-alt='gsea_f_myonuclei_cac_vs_ctrl.png'}

### gsea_f_myonuclei_cac_vs_pre.png

![](assets/figures/gsea/gsea_f_myonuclei_cac_vs_pre.png){fig-alt='gsea_f_myonuclei_cac_vs_pre.png'}

### gsea_f_myonuclei_pre_vs_ctrl.png

![](assets/figures/gsea/gsea_f_myonuclei_pre_vs_ctrl.png){fig-alt='gsea_f_myonuclei_pre_vs_ctrl.png'}

### gsea_m_myonuclei_cac_vs_ctrl.png

![](assets/figures/gsea/gsea_m_myonuclei_cac_vs_ctrl.png){fig-alt='gsea_m_myonuclei_cac_vs_ctrl.png'}

### gsea_m_myonuclei_cac_vs_pre.png

![](assets/figures/gsea/gsea_m_myonuclei_cac_vs_pre.png){fig-alt='gsea_m_myonuclei_cac_vs_pre.png'}

### gsea_m_myonuclei_pre_vs_ctrl.png

![](assets/figures/gsea/gsea_m_myonuclei_pre_vs_ctrl.png){fig-alt='gsea_m_myonuclei_pre_vs_ctrl.png'}

### qc_K017_atac.jpg

![](assets/figures/qc/qc_K017_atac.jpg){fig-alt='qc_K017_atac.jpg'}

### qc_K017_filtered.jpg

![](assets/figures/qc/qc_K017_filtered.jpg){fig-alt='qc_K017_filtered.jpg'}

### qc_K017_rna.jpg

![](assets/figures/qc/qc_K017_rna.jpg){fig-alt='qc_K017_rna.jpg'}

### qc_K017_unfiltered.jpg

![](assets/figures/qc/qc_K017_unfiltered.jpg){fig-alt='qc_K017_unfiltered.jpg'}

Legacy Presentations

The old slide decks stay outside the Git clone for now. This table indexes the local presentation archive so the useful figures can be migrated gradually into this Quarto document.

File Type Size_MB Modified
46 14_Presentazione_13_04/Presentazione_02-03-26.pptx PPTX 40.7 2026-04-13
45 13_Presentrazione_23-03/Presentazione_02-03-26.pptx PPTX 9.1 2026-03-22
44 12_Presentazione_02-03-26/Presentazione_02-03-26.pptx PPTX 15.9 2026-03-03
43 12_Presentazione_02-03-26/Presentazione_02-03-26.pdf PDF 7.6 2026-03-02
58 5-Presentazione-02-10-25/5-Presentazione_02-10-2025.pptx PPTX 21.1 2026-02-26
42 11_Presentazione_11-02-26/Presentazione_11-02-26.pptx PPTX 10.9 2026-02-23
41 11_Presentazione_11-02-26/Presentazione_11-02-26.pdf PDF 6.1 2026-02-11
70 Annotazione.pptx PPTX 2.8 2026-02-09
40 10_Presentazione_22-01-26/Presentazione_22-01-26.pptx PPTX 3.5 2026-01-21
66 9_Presentazione_Francesi-08-01-26/08-01.pptx PPTX 11.0 2026-01-08
69 9_Presentazione_Francesi-08-01-26/Summary.pdf PDF 0.1 2026-01-07
68 9_Presentazione_Francesi-08-01-26/Script_Presentazione.pdf PDF 0.1 2026-01-07
67 9_Presentazione_Francesi-08-01-26/QC.pdf PDF 0.1 2026-01-07
65 8_Presentazione_18-12-25/8-Presentazione_18-12-2025.pptx PPTX 5.1 2025-12-17
64 7_Presentazione_04-12-25/7-Presentazione_04-12-2025.pptx PPTX 58.1 2025-12-04
63 7_Presentazione_04-12-25/7-CellChat_04-12-2025.pptx PPTX 32.2 2025-12-01
84 Extra/6-Stabilita_Cluster.pdf PDF 3.0 2025-11-06
61 6-Presentazione-06-11-25/6-Presentazione_OLD_06-11-2025.pptx PPTX 107.1 2025-11-06
60 6-Presentazione-06-11-25/6-Presentazione_06-11-2025.pptx PPTX 63.0 2025-11-06
59 6-Presentazione-06-11-25/6-Presentazione_06-11-2025.pdf PDF 34.9 2025-11-06
85 Extra/6-Stabilita_Cluster.pptx PPTX 4.2 2025-10-27
62 6-Presentazione-06-11-25/6-Scriabin.pptx PPTX 2.1 2025-10-27
57 5-Presentazione-02-10-25/5-Presentazione_02-10-2025.pdf PDF 14.7 2025-10-02
82 Extra/2_01-08/GSEA_Male_ComBat/2_GSEA_Male_31-07.pptx PPTX 3.9 2025-08-02
81 Extra/2_01-08/GSEA_Male_ComBat/2_GSEA_Male.pdf PDF 3.3 2025-08-02
79 Extra/2_01-08/GSEA_Female_RUV/No120/2_GSEA_Female_No120.pdf PDF 4.2 2025-08-02
80 Extra/2_01-08/GSEA_Female_RUV/No120/2_GSEA_No120_31-07.pptx PPTX 5.0 2025-08-01
77 Extra/2_01-08/GSEA_Female_RUV/Best/2_GSEA_Female_No120_No63.pdf PDF 4.0 2025-08-01
74 Extra/2_01-08/1_DEGs_IndagineCombinazioni_Summary.pdf PDF 0.8 2025-08-01
73 Extra/2_01-08/1_DEGs_IndagineCombinazioni.pdf PDF 8.4 2025-08-01
Copied asset manifest
Section Asset Source
QC qc/qc_K017_unfiltered.jpg 0_QC_graph_new/S65235_K017_QC_unfiltered.jpg
QC qc/qc_K017_filtered.jpg 0_QC_graph_new/S65235_K017_QC_filtered.jpg
QC qc/qc_K017_rna.jpg 0_QC_graph_new/S65235_K017_QC_RNA.jpg
QC qc/qc_K017_atac.jpg 0_QC_graph_new/S65235_K017_QC_ATAC.jpg
Annotation annotation/annotation_f_umap_combined.png 3_Ann_pt2/F/UMAP_annot_combined.png
Annotation annotation/annotation_f_marker_dotplot.png 3_Ann_pt2/F/Multiome_DotPlot_Markers.png
Annotation annotation/annotation_f_cells_by_condition.png 3_Ann_pt2/F/BarPlot_PerCellbyCondition.png
Annotation annotation/annotation_m_umap_combined.png 3_Ann_pt2/M/UMAP_annot_combined.png
Annotation annotation/annotation_m_marker_dotplot.png 3_Ann_pt2/M/Multiome_DotPlot_Markers.png
Annotation annotation/annotation_m_cells_by_condition.png 3_Ann_pt2/M/BarPlot_PerCellbyCondition.png
GSEA gsea/gsea_f_myonuclei_cac_vs_ctrl.png 8_GSEA_New/F/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png
GSEA gsea/gsea_f_myonuclei_cac_vs_pre.png 8_GSEA_New/F/MyoOverall_CACvsPRE_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_PRE.png
GSEA gsea/gsea_f_myonuclei_pre_vs_ctrl.png 8_GSEA_New/F/MyoOverall_PREvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_PRE_vs_CTRL.png
GSEA gsea/gsea_m_myonuclei_cac_vs_ctrl.png 8_GSEA/M/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png
GSEA gsea/gsea_m_myonuclei_cac_vs_pre.png 8_GSEA/M/MyoOverall_CACvsPRE_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_PRE.png
GSEA gsea/gsea_m_myonuclei_pre_vs_ctrl.png 8_GSEA/M/MyoOverall_PREvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_PRE_vs_CTRL.png
DEVIL GSEA devil/devil_gsea_f_myonuclei_cac_vs_ctrl.png 8_GSEA_devil_New/F/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png
DEVIL GSEA devil/devil_gsea_m_myonuclei_cac_vs_ctrl.png 8_GSEA_devil_New/M/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png
CCC / Scriabin ccc/ccc_myoIIx_to_tcells_clusters.png EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells/CCIM_UMAP_by_clusters.png
CCC / Scriabin ccc/ccc_myoIIx_to_tcells_sender.png EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells/CCIM_UMAP_by_sender_clusters.png
CCC / Scriabin ccc/ccc_myoIIx_to_tcells_receiver.png EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells/CCIM_UMAP_by_receiver_clusters.png
CCC / Scriabin ccc/ccc_myoIIx_to_tcells_lr_dotplot.png EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells_markers/DotPlot_topLR_cluster0.png

Repository Map

Top_level
0_tbi_file.R
0_tbi_file_NewData.R
1_QualityControl.R
1_QualityControl_NewData.R
2_RemoveDoublets_Fem.R
2_RemoveDoublets_Fem_NewData.R
2_RemoveDoublets_Man.R
2_RemoveDoublets_Man_NewData.R
3_QC_ribosom.R
hdf5-1.14.3.tar.gz
NuovaPipeNo120
Output
Poster
quarto_interactive
All R scripts currently tracked in the analysis folders
File
0_tbi_file.R
0_tbi_file_NewData.R
1_QualityControl.R
1_QualityControl_NewData.R
2_RemoveDoublets_Fem.R
2_RemoveDoublets_Fem_NewData.R
2_RemoveDoublets_Man.R
2_RemoveDoublets_Man_NewData.R
3_QC_ribosom.R
NuovaPipeNo120/0_PseudoBulkxClusterF.R
NuovaPipeNo120/0_PseudoBulkxClusterM.R
NuovaPipeNo120/1_IntFem_NewData.R
NuovaPipeNo120/1_IntMan_NewData.R
NuovaPipeNo120/10_CCC_120_pt1.R
NuovaPipeNo120/10_CCC_120_pt2.R
NuovaPipeNo120/10_CCC_F.R
NuovaPipeNo120/10_CCC_F_perPaz.R
NuovaPipeNo120/10_CCC_M.R
NuovaPipeNo120/10_CCC_M_perPaz.R
NuovaPipeNo120/2_AnnFem_New.R
NuovaPipeNo120/2_AnnMan_New.R
NuovaPipeNo120/3_AnnFem_pt2_New.R
NuovaPipeNo120/3_AnnMan_pt2_New.R
NuovaPipeNo120/4_FiltGenes_F_NewData.R
NuovaPipeNo120/4_FiltGenes_M_NewData.R
NuovaPipeNo120/5_DEG_F_New.R
NuovaPipeNo120/5_DEG_M_New.R
NuovaPipeNo120/5_DEG_New.R
NuovaPipeNo120/6_DEA_devil_female.R
NuovaPipeNo120/6_DEA_devil_male.R
NuovaPipeNo120/7_GSEA_F.R
NuovaPipeNo120/7_GSEA_M_New.R
NuovaPipeNo120/8_GSEA_F_Dev.R
NuovaPipeNo120/8_GSEA_M_Dev.R
NuovaPipeNo120/8_Ribosomal_PCA_Heatmaps_MF.R
NuovaPipeNo120/ConfrontoLabelMan.R
NuovaPipeNo120/Dotplot.R
NuovaPipeNo120/Dotplot_F.R
NuovaPipeNo120/Dotplot_M.R
NuovaPipeNo120/Heatmap_F.R
NuovaPipeNo120/Heatmap_M.R
NuovaPipeNo120/HeatRibo.R
NuovaPipeNo120/old/0_bulkPCA_All.R
NuovaPipeNo120/old/0_bulkPCA_All_No120.R
NuovaPipeNo120/old/0_bulkPCA_F.R
NuovaPipeNo120/old/0_bulkPCA_F_NewData_NoC063.R
NuovaPipeNo120/old/0_bulkPCA_F_No120.R
NuovaPipeNo120/old/0_bulkPCA_M.R
NuovaPipeNo120/old/1_IntFem.R
NuovaPipeNo120/old/1_IntMan.R
NuovaPipeNo120/old/2_AnnFem.R
NuovaPipeNo120/old/2_AnnMan.R
NuovaPipeNo120/old/3_AnnFem_pt2.R
NuovaPipeNo120/old/3_AnnMan_pt2.R
NuovaPipeNo120/old/4_FiltGenes_F.R
NuovaPipeNo120/old/4_FiltGenes_M.R
NuovaPipeNo120/old/5_DEG_F.R
NuovaPipeNo120/old/5_DEG_M.R
NuovaPipeNo120/old/5_DEGs_M_skeletal.R
NuovaPipeNo120/old/5_DEGs_M_Undet.R
NuovaPipeNo120/old/9_GSEA_M.R
NuovaPipeNo120/ribo/EXTRA_Ribo_Dotplot_F.R
NuovaPipeNo120/ribo/EXTRA_Ribo_Dotplot_M.R
NuovaPipeNo120/ribo/EXTRA_Ribo_PCA_F.R
NuovaPipeNo120/ribo/EXTRA_Ribo_PCA_M.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt1.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt2.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt2_Global.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt2_NoGrafo.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt3.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt3_Global.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt4.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CAC_vs_CTRL_pt4_Global.R
NuovaPipeNo120/scriabin/11_Scriabin_M_CTRL_vs_CAC_pt2.R
NuovaPipeNo120/scriabin/11_Scriabin_M_FiltGenes_pt2.R
NuovaPipeNo120/scriabin/13_CCIM_CellChatStyle_MyoIIx_to_TCells.R
NuovaPipeNo120/Umap_F.R
NuovaPipeNo120/Umap_M.R
NuovaPipeNo120/Upset_M.R
Poster/Annotazioni/clust2.R
Poster/Annotazioni/clust3.R
Poster/Annotazioni/CreateClusterExp.R
Poster/Annotazioni/provaClust.R
Poster/Annotazioni/provaClust2.R
Poster/Annotazioni/Seurat.R
Poster/Annotazioni/sgd.R
Poster/confronto_Fem.R
Poster/Seurat/1_IntFem_seurat_20k-1k-10.R
Poster/Seurat/1_IntFem_seurat_20k-1k-20.R
Poster/Seurat/1_IntFem_seurat_20k-1k-30.R
Poster/Seurat/1_IntFem_seurat_20k-2k-10.R
Poster/Seurat/1_IntFem_seurat_20k-2k-20.R
Poster/Seurat/1_IntFem_seurat_20k-2k-30.R
Poster/Seurat/1_IntFem_seurat_20k-3k-10.R
Poster/Seurat/1_IntFem_seurat_20k-3k-20.R
Poster/Seurat/1_IntFem_seurat_20k-3k-30.R
Poster/Seurat/1_IntFem_seurat_50k-1k-10.R
Poster/Seurat/1_IntFem_seurat_50k-1k-20.R
Poster/Seurat/1_IntFem_seurat_50k-1k-30.R
Poster/Seurat/1_IntFem_seurat_50k-2k-10.R
Poster/Seurat/1_IntFem_seurat_50k-2k-20.R
Poster/Seurat/1_IntFem_seurat_50k-2k-30.R
Poster/Seurat/1_IntFem_seurat_50k-3k-10.R
Poster/Seurat/1_IntFem_seurat_50k-3k-20.R
Poster/Seurat/1_IntFem_seurat_50k-3k-30.R
Poster/Seurat/1_IntFem_seurat_Max-1k-10.R
Poster/Seurat/1_IntFem_seurat_Max-1k-20.R
Poster/Seurat/1_IntFem_seurat_Max-1k-30.R
Poster/Seurat/1_IntFem_seurat_Max-2k-10.R
Poster/Seurat/1_IntFem_seurat_Max-2k-20.R
Poster/Seurat/1_IntFem_seurat_Max-2k-30.R
Poster/Seurat/1_IntFem_seurat_Max-3k-10.R
Poster/Seurat/1_IntFem_seurat_Max-3k-20.R
Poster/Seurat/1_IntFem_seurat_Max-3k-30.R
Poster/sgdGMF/annotazione/0_Preprocessing_Integration_Fem.R
Poster/sgdGMF/annotazione/0_Preprocessing_Integration_M.R
Poster/sgdGMF/annotazione/2_IntFem_30.R
Poster/sgdGMF/annotazione/2_IntM_30.R
Poster/sgdGMF/annotazione/3_Ann_M_30.R
Poster/sgdGMF/annotazione/3_AnnFem30.R
Poster/sgdGMF/annotazione/4_AnnFem_pt2.R
Poster/sgdGMF/sgd_20k-1k-10.R
Poster/sgdGMF/sgd_20k-1k-20.R
Poster/sgdGMF/sgd_20k-1k-30.R
Poster/sgdGMF/sgd_20k-2k-10.R
Poster/sgdGMF/sgd_20k-2k-20.R
Poster/sgdGMF/sgd_20k-2k-30.R
Poster/sgdGMF/sgd_20k-3k-10.R
Poster/sgdGMF/sgd_20k-3k-20.R
Poster/sgdGMF/sgd_20k-3k-30.R
Poster/sgdGMF/sgd_50k-1k-10.R
Poster/sgdGMF/sgd_50k-1k-20.R
Poster/sgdGMF/sgd_50k-1k-30.R
Poster/sgdGMF/sgd_50k-2k-10.R
Poster/sgdGMF/sgd_50k-2k-20.R
Poster/sgdGMF/sgd_50k-2k-30.R
Poster/sgdGMF/sgd_50k-3k-10.R
Poster/sgdGMF/sgd_50k-3k-20.R
Poster/sgdGMF/sgd_50k-3k-30.R
Poster/sgdGMF/sgd_Max-1k-10.R
Poster/sgdGMF/sgd_Max-1k-20.R
Poster/sgdGMF/sgd_Max-1k-30.R
Poster/sgdGMF/sgd_Max-2k-10.R
Poster/sgdGMF/sgd_Max-2k-20.R
Poster/sgdGMF/sgd_Max-2k-30.R
Poster/sgdGMF/sgd_Max-3k-10.R
Poster/sgdGMF/sgd_Max-3k-20.R
Poster/sgdGMF/sgd_Max-3k-30.R

Discussion Questions

QC

Which samples or clusters are most sensitive to the filtering choices?

Annotation

Which cell-state labels are strong enough for the main story, and which should remain provisional?

Sex-specificity

Which signals replicate in female and male analyses, and which are biologically plausible as sex-specific?

Mechanism

Which pathway or communication signal deserves validation first?

Meeting Notes

Use this section during live discussion.

  • Main concern raised:
  • Figure requested:
  • Follow-up analysis:
  • Decision:
Source Code
---
title: "Cachectic Multiome Human"
subtitle: "Interactive discussion document"
author: "Sophie Grace Parolin"
date: last-modified
format:
  html:
    self-contained: false
---

```{r setup}
library(knitr)

repo_root <- normalizePath("..", winslash = "/", mustWork = TRUE)
archive_root <- normalizePath("../..", winslash = "/", mustWork = FALSE)

list_repo_files <- function(pattern, root = repo_root) {
  files <- list.files(root, pattern = pattern, recursive = TRUE, full.names = FALSE)
  sort(files)
}

legacy_presentations <- function() {
  pdir <- file.path(archive_root, "Presentazioni")
  if (!dir.exists(pdir)) {
    return(data.frame())
  }

  files <- list.files(
    pdir,
    pattern = "\\.(pptx|ppt|pdf)$",
    recursive = TRUE,
    full.names = TRUE
  )

  info <- file.info(files)
  out <- data.frame(
    File = gsub(paste0(normalizePath(pdir, winslash = "/", mustWork = TRUE), "/"), "", normalizePath(files, winslash = "/", mustWork = FALSE), fixed = TRUE),
    Type = toupper(sub(".*\\.", "", files)),
    Size_MB = round(info$size / 1024^2, 1),
    Modified = as.Date(info$mtime),
    stringsAsFactors = FALSE
  )
  out[order(out$Modified, out$File, decreasing = TRUE), ]
}

script_table <- data.frame(
  Step = c(
    "0",
    "1",
    "2",
    "3",
    "4",
    "5",
    "6",
    "7-8",
    "10"
  ),
  Module = c(
    "Input metadata / TBI files",
    "Integration by sex",
    "First annotation",
    "Refined annotation",
    "Gene filtering",
    "Pseudo-bulk differential expression",
    "Cell-level DE with devil",
    "GSEA / pathway interpretation",
    "Cell-cell communication"
  ),
  Key_files = c(
    "`0_tbi_file*.R`",
    "`NuovaPipeNo120/1_Int*_NewData.R`",
    "`NuovaPipeNo120/2_Ann*_New.R`",
    "`NuovaPipeNo120/3_Ann*_pt2_New.R`",
    "`NuovaPipeNo120/4_FiltGenes_*_NewData.R`",
    "`NuovaPipeNo120/5_DEG*.R`",
    "`NuovaPipeNo120/6_DEA_devil_*.R`",
    "`NuovaPipeNo120/7_GSEA_F.R`, `8_GSEA_*`",
    "`NuovaPipeNo120/10_CCC_*.R`"
  )
)

figures <- list.files("assets/figures", pattern = "\\.(png|jpg|jpeg|svg)$", recursive = TRUE, full.names = TRUE)
copied_assets <- data.frame(
  Section = c(
    rep("QC", 4),
    rep("Annotation", 6),
    rep("GSEA", 6),
    rep("DEVIL GSEA", 2),
    rep("CCC / Scriabin", 4)
  ),
  Asset = c(
    "qc/qc_K017_unfiltered.jpg",
    "qc/qc_K017_filtered.jpg",
    "qc/qc_K017_rna.jpg",
    "qc/qc_K017_atac.jpg",
    "annotation/annotation_f_umap_combined.png",
    "annotation/annotation_f_marker_dotplot.png",
    "annotation/annotation_f_cells_by_condition.png",
    "annotation/annotation_m_umap_combined.png",
    "annotation/annotation_m_marker_dotplot.png",
    "annotation/annotation_m_cells_by_condition.png",
    "gsea/gsea_f_myonuclei_cac_vs_ctrl.png",
    "gsea/gsea_f_myonuclei_cac_vs_pre.png",
    "gsea/gsea_f_myonuclei_pre_vs_ctrl.png",
    "gsea/gsea_m_myonuclei_cac_vs_ctrl.png",
    "gsea/gsea_m_myonuclei_cac_vs_pre.png",
    "gsea/gsea_m_myonuclei_pre_vs_ctrl.png",
    "devil/devil_gsea_f_myonuclei_cac_vs_ctrl.png",
    "devil/devil_gsea_m_myonuclei_cac_vs_ctrl.png",
    "ccc/ccc_myoIIx_to_tcells_clusters.png",
    "ccc/ccc_myoIIx_to_tcells_sender.png",
    "ccc/ccc_myoIIx_to_tcells_receiver.png",
    "ccc/ccc_myoIIx_to_tcells_lr_dotplot.png"
  ),
  Source = c(
    "0_QC_graph_new/S65235_K017_QC_unfiltered.jpg",
    "0_QC_graph_new/S65235_K017_QC_filtered.jpg",
    "0_QC_graph_new/S65235_K017_QC_RNA.jpg",
    "0_QC_graph_new/S65235_K017_QC_ATAC.jpg",
    "3_Ann_pt2/F/UMAP_annot_combined.png",
    "3_Ann_pt2/F/Multiome_DotPlot_Markers.png",
    "3_Ann_pt2/F/BarPlot_PerCellbyCondition.png",
    "3_Ann_pt2/M/UMAP_annot_combined.png",
    "3_Ann_pt2/M/Multiome_DotPlot_Markers.png",
    "3_Ann_pt2/M/BarPlot_PerCellbyCondition.png",
    "8_GSEA_New/F/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png",
    "8_GSEA_New/F/MyoOverall_CACvsPRE_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_PRE.png",
    "8_GSEA_New/F/MyoOverall_PREvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_PRE_vs_CTRL.png",
    "8_GSEA/M/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png",
    "8_GSEA/M/MyoOverall_CACvsPRE_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_PRE.png",
    "8_GSEA/M/MyoOverall_PREvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_PRE_vs_CTRL.png",
    "8_GSEA_devil_New/F/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png",
    "8_GSEA_devil_New/M/MyoOverall_CACvsCTRL_WithRibo/dotplots/GSEA_dotplot_Myonuclei_CAC_vs_CTRL.png",
    "EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells/CCIM_UMAP_by_clusters.png",
    "EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells/CCIM_UMAP_by_sender_clusters.png",
    "EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells/CCIM_UMAP_by_receiver_clusters.png",
    "EXTRA/11_CCC_Scriabin/CCIM_CAC_Myo_IIx_to_TCells_markers/DotPlot_topLR_cluster0.png"
  )
)
```

::: {.hero-strip}
::: {.talk-card}
**Core aim**

Use this page as the live presentation surface for the cachexia multiome project: start with the main biological story, then jump into QC, annotation, DEG, GSEA, DEVIL, or CCC only when someone asks.

**Status**

This is the scaffold. It is ready for discussion notes and exported figures; the analysis scripts remain in the repository as the reproducible backend.
:::

::: {.route-map}
**Jump to what they ask**

[Study map](#study-map)
[QC](#quality-control)
[Annotation](#annotation)
[DEG](#differential-expression)
[DEVIL](#devil-cell-level-de)
[GSEA](#gsea-and-pathways)
[CCC](#cell-cell-communication)
[Files](#repository-map)
[Open questions](#discussion-questions)
:::
:::

## Study Map

::: {.callout-note}
## Presentation principle

Keep each section to one answerable question, one primary visual, and one collapsible methods block. During a meeting, the document becomes a map rather than a long linear talk.
:::

```{r}
kable(script_table, escape = FALSE)
```

<details>
<summary>Talking path for a 5-minute overview</summary>

1. What biological contrast defines cachexia in this cohort?
2. Which nuclei/cell states are robust after QC and annotation?
3. Which changes are shared versus sex-specific?
4. Which pathways move consistently across pseudo-bulk, DEVIL, and GSEA?
5. Which cell-cell communication signals are worth validating?

</details>

## Quality Control

**Question to answer live:** are the nuclei/cells reliable enough to support the downstream story?

::: {.figure-grid}
::: {.figure-card}
**K017 before filtering**

![](assets/figures/qc/qc_K017_unfiltered.jpg)
:::

::: {.figure-card}
**K017 after filtering**

![](assets/figures/qc/qc_K017_filtered.jpg)
:::

::: {.figure-card}
**RNA QC**

![](assets/figures/qc/qc_K017_rna.jpg)
:::

::: {.figure-card}
**ATAC QC**

![](assets/figures/qc/qc_K017_atac.jpg)
:::
:::

<details>
<summary>Relevant scripts and logs</summary>

```{r}
qc_files <- c(
  list_repo_files("^1_QualityControl.*\\.R$"),
  list_repo_files("^2_RemoveDoublets_.*\\.R$"),
  list_repo_files("^3_QC_ribosom.*\\.R$"),
  list_repo_files("^Output/.*\\.Rout$")
)
kable(data.frame(File = qc_files))
```

</details>

## Annotation

**Question to answer live:** what cell types/states are present, and which annotations are stable across RNA, ATAC, and WNN views?

::: {.panel-tabset}

### Female

::: {.figure-grid}
::: {.figure-card}
**Annotated UMAP**

![](assets/figures/annotation/annotation_f_umap_combined.png)
:::

::: {.figure-card}
**Marker dotplot**

![](assets/figures/annotation/annotation_f_marker_dotplot.png)
:::

::: {.figure-card}
**Cell composition by condition**

![](assets/figures/annotation/annotation_f_cells_by_condition.png)
:::
:::

### Male

::: {.figure-grid}
::: {.figure-card}
**Annotated UMAP**

![](assets/figures/annotation/annotation_m_umap_combined.png)
:::

::: {.figure-card}
**Marker dotplot**

![](assets/figures/annotation/annotation_m_marker_dotplot.png)
:::

::: {.figure-card}
**Cell composition by condition**

![](assets/figures/annotation/annotation_m_cells_by_condition.png)
:::
:::

### Marker Logic

Use this tab for the curated marker list and difficult annotation decisions.

:::

<details>
<summary>Relevant scripts</summary>

```{r}
ann_files <- list_repo_files("NuovaPipeNo120/(2_Ann|3_Ann|Dotplot|Heatmap|Umap).*\\.R$")
kable(data.frame(File = ann_files))
```

</details>

## Differential Expression

**Question to answer live:** which transcriptional changes separate CAC, PRE, and CTRL, and are they cluster-specific or global?

::: {.panel-tabset}

### Pseudo-bulk

The current local archive contains DEG results mainly as `.xlsx` tables; this tab keeps their locations close to the visual pathway summaries below.

```{r}
deg_tables <- list.files(
  file.path(archive_root, "5_DEG_10"),
  pattern = "\\.xlsx$",
  recursive = TRUE,
  full.names = FALSE
)
kable(data.frame(Table = sort(deg_tables)))
```

### Female

Use this tab for CAC vs CTRL, CAC vs PRE, and PRE vs CTRL in female samples.

### Male

Use this tab for CAC vs CTRL, CAC vs PRE, and PRE vs CTRL in male samples.

### Myonuclei

Use this tab for myonuclei-specific contrasts and comparisons with global/cluster-level results.

:::

<details>
<summary>Relevant scripts and outputs</summary>

```{r}
deg_files <- c(
  list_repo_files("NuovaPipeNo120/5_DEG.*\\.R$"),
  list_repo_files("NuovaPipeNo120/Output/.*DEG.*\\.Rout$")
)
kable(data.frame(File = deg_files))
```

</details>

## DEVIL Cell-Level DE

**Question to answer live:** do cell-level models support or refine the pseudo-bulk signals?

```{r}
devil_tables <- list.files(
  file.path(archive_root, "5_DEG_devil"),
  pattern = "\\.xlsx$",
  recursive = TRUE,
  full.names = FALSE
)
kable(data.frame(Table = sort(devil_tables)))
```

::: {.small-note}
The visual DEVIL-ranked pathway summaries are shown under [GSEA and pathways](#gsea-and-pathways), in the DEVIL-ranked GSEA tab.
:::

<details>
<summary>Relevant scripts and logs</summary>

```{r}
devil_files <- c(
  list_repo_files("NuovaPipeNo120/6_DEA_devil.*\\.R$"),
  list_repo_files("NuovaPipeNo120/6_DEA_devil.*\\.Rout$")
)
kable(data.frame(File = devil_files))
```

</details>

## GSEA And Pathways

**Question to answer live:** which pathways make the gene-level changes biologically coherent?

::: {.panel-tabset}

### Female GSEA

::: {.figure-grid}
::: {.figure-card}
**Myonuclei CAC vs CTRL**

![](assets/figures/gsea/gsea_f_myonuclei_cac_vs_ctrl.png)
:::

::: {.figure-card}
**Myonuclei CAC vs PRE**

![](assets/figures/gsea/gsea_f_myonuclei_cac_vs_pre.png)
:::

::: {.figure-card}
**Myonuclei PRE vs CTRL**

![](assets/figures/gsea/gsea_f_myonuclei_pre_vs_ctrl.png)
:::
:::

### Male GSEA

::: {.figure-grid}
::: {.figure-card}
**Myonuclei CAC vs CTRL**

![](assets/figures/gsea/gsea_m_myonuclei_cac_vs_ctrl.png)
:::

::: {.figure-card}
**Myonuclei CAC vs PRE**

![](assets/figures/gsea/gsea_m_myonuclei_cac_vs_pre.png)
:::

::: {.figure-card}
**Myonuclei PRE vs CTRL**

![](assets/figures/gsea/gsea_m_myonuclei_pre_vs_ctrl.png)
:::
:::

### DEVIL-ranked GSEA

::: {.figure-grid}
::: {.figure-card}
**Female myonuclei CAC vs CTRL**

![](assets/figures/devil/devil_gsea_f_myonuclei_cac_vs_ctrl.png)
:::

::: {.figure-card}
**Male myonuclei CAC vs CTRL**

![](assets/figures/devil/devil_gsea_m_myonuclei_cac_vs_ctrl.png)
:::
:::

:::

<details>
<summary>Relevant scripts and logs</summary>

```{r}
gsea_files <- c(
  list_repo_files("NuovaPipeNo120/[78]_GSEA.*\\.R$"),
  list_repo_files("NuovaPipeNo120/Output/8_GSEA.*\\.Rout$")
)
kable(data.frame(File = gsea_files))
```

</details>

## Cell-Cell Communication

**Question to answer live:** which ligand-receptor programs connect cachexia-associated states?

::: {.figure-grid}
::: {.figure-card}
**CAC Myo IIx to T cells: clusters**

![](assets/figures/ccc/ccc_myoIIx_to_tcells_clusters.png)
:::

::: {.figure-card}
**Sender clusters**

![](assets/figures/ccc/ccc_myoIIx_to_tcells_sender.png)
:::

::: {.figure-card}
**Receiver clusters**

![](assets/figures/ccc/ccc_myoIIx_to_tcells_receiver.png)
:::

::: {.figure-card}
**Top ligand-receptor markers**

![](assets/figures/ccc/ccc_myoIIx_to_tcells_lr_dotplot.png)
:::
:::

<details>
<summary>Relevant scripts and logs</summary>

```{r}
ccc_files <- c(
  list_repo_files("NuovaPipeNo120/10_CCC.*\\.R$"),
  list_repo_files("NuovaPipeNo120/scriabin/.*\\.R$"),
  list_repo_files("NuovaPipeNo120/Output/9_CCC.*\\.Rout$")
)
kable(data.frame(File = ccc_files))
```

</details>

## Figure Gallery

```{r}
if (length(figures) == 0) {
  cat('<div class="figure-slot">No figures found yet in <code>quarto_interactive/assets/figures/</code>. Export selected PNG/JPG/SVG files there and re-render.</div>')
} else {
  for (fig in figures) {
    cat(sprintf("### %s\n\n![](%s){fig-alt='%s'}\n\n", basename(fig), fig, basename(fig)))
  }
}
```

## Legacy Presentations

The old slide decks stay outside the Git clone for now. This table indexes the local presentation archive so the useful figures can be migrated gradually into this Quarto document.

```{r}
legacy <- legacy_presentations()
if (nrow(legacy) == 0) {
  cat('<div class="figure-slot">No local <code>Presentazioni/</code> folder found next to the cloned repository.</div>')
} else {
  kable(head(legacy, 30))
}
```

<details>
<summary>Copied asset manifest</summary>

```{r}
kable(copied_assets)
```

</details>

## Repository Map

```{r}
top_level <- list.files(repo_root, all.files = FALSE, recursive = FALSE)
kable(data.frame(Top_level = sort(top_level)))
```

<details>
<summary>All R scripts currently tracked in the analysis folders</summary>

```{r}
all_r <- list_repo_files("\\.R$")
kable(data.frame(File = all_r))
```

</details>

## Discussion Questions

::: {.question-bank}
<div>
**QC**

Which samples or clusters are most sensitive to the filtering choices?
</div>

<div>
**Annotation**

Which cell-state labels are strong enough for the main story, and which should remain provisional?
</div>

<div>
**Sex-specificity**

Which signals replicate in female and male analyses, and which are biologically plausible as sex-specific?
</div>

<div>
**Mechanism**

Which pathway or communication signal deserves validation first?
</div>
:::

## Meeting Notes

Use this section during live discussion.

- Main concern raised:
- Figure requested:
- Follow-up analysis:
- Decision: