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[ci skip] This build is based on 7e4d905. This commit was created by the following CI build and job: https://github.com/AlexsLemonade/ScPCA-manuscript/commit/7e4d9053d8d1d2619806ef331d98138dededdac8/checks https://github.com/AlexsLemonade/ScPCA-manuscript/actions/runs/20959556332
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citations.tsv

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doi:10.1016/j.celrep.2024.114804 doi:10.1016/j.celrep.2024.114804 doi:10.1016/j.celrep.2024.114804 sGIqMPJO
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url:https://github.com/broadinstitute/inferCNV url:https://github.com/broadinstitute/inferCNV url:https://github.com/broadinstitute/inferCNV b9hQbZOR
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doi:10.1038/nrdp.2016.78 doi:10.1038/nrdp.2016.78 doi:10.1038/nrdp.2016.78 11NujsTQW
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doi:10.1158/2159-8290.CD-14-0622 doi:10.1158/2159-8290.CD-14-0622 doi:10.1158/2159-8290.cd-14-0622 1Ar16jCVg
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doi:10.3389/fphar.2024.1355242 doi:10.3389/fphar.2024.1355242 doi:10.3389/fphar.2024.1355242 DR9mGrXH
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url:https://openscpca.readthedocs.io url:https://openscpca.readthedocs.io url:https://openscpca.readthedocs.io xW9vfui5
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url:https://github.com/AlexsLemonade/OpenScPCA-analysis url:https://github.com/AlexsLemonade/OpenScPCA-analysis url:https://github.com/AlexsLemonade/OpenScPCA-analysis ckn7Y4Ch
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url:https://github.com/AlexsLemonade/OpenScPCA-nf url:https://github.com/AlexsLemonade/OpenScPCA-nf url:https://github.com/AlexsLemonade/OpenScPCA-nf tDa8M4VA
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url:https://github.com/broadinstitute/inferCNV url:https://github.com/broadinstitute/inferCNV url:https://github.com/broadinstitute/inferCNV b9hQbZOR
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doi:10.1038/nbt.4091 doi:10.1038/nbt.4091 doi:10.1038/nbt.4091 kILCvszy
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doi:10.1016/j.xgen.2023.100340 doi:10.1016/j.xgen.2023.100340 doi:10.1016/j.xgen.2023.100340 5VXMHJ7N
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doi:10.1101/2024.01.07.574538 doi:10.1101/2024.01.07.574538 doi:10.1101/2024.01.07.574538 TSJacwEL
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doi:10.1186/s13059-024-03309-4 doi:10.1186/s13059-024-03309-4 doi:10.1186/s13059-024-03309-4 1G3mGXjU5
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doi:10.1038/nrdp.2016.78 doi:10.1038/nrdp.2016.78 doi:10.1038/nrdp.2016.78 11NujsTQW
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doi:10.1016/j.celrep.2024.114804 doi:10.1016/j.celrep.2024.114804 doi:10.1016/j.celrep.2024.114804 sGIqMPJO
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doi:10.1158/2159-8290.CD-14-0622 doi:10.1158/2159-8290.CD-14-0622 doi:10.1158/2159-8290.cd-14-0622 1Ar16jCVg

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- open science
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- reproducibility
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lang: en-US
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date-meta: '2026-01-12'
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date-meta: '2026-01-13'
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author-meta:
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- Allegra G. Hawkins
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- Joshua A. Shapiro
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<meta name="citation_title" content="The Single-cell Pediatric Cancer Atlas: Data portal and open-source tools for single-cell transcriptomics of pediatric tumors" />
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<meta property="og:title" content="The Single-cell Pediatric Cancer Atlas: Data portal and open-source tools for single-cell transcriptomics of pediatric tumors" />
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<meta property="twitter:title" content="The Single-cell Pediatric Cancer Atlas: Data portal and open-source tools for single-cell transcriptomics of pediatric tumors" />
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<meta name="dc.date" content="2026-01-12" />
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<small><em>
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This manuscript
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([permalink](https://AlexsLemonade.github.io/ScPCA-manuscript/v/0d5913b9256ff4a452929c68030d05ab8f71b2ea/))
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([permalink](https://AlexsLemonade.github.io/ScPCA-manuscript/v/7e4d9053d8d1d2619806ef331d98138dededdac8/))
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was automatically generated
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from [AlexsLemonade/ScPCA-manuscript@0d5913b](https://github.com/AlexsLemonade/ScPCA-manuscript/tree/0d5913b9256ff4a452929c68030d05ab8f71b2ea)
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on January 12, 2026.
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from [AlexsLemonade/ScPCA-manuscript@7e4d905](https://github.com/AlexsLemonade/ScPCA-manuscript/tree/7e4d9053d8d1d2619806ef331d98138dededdac8)
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on January 13, 2026.
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</em></small>
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Figure {@fig:fig4}D shows the expression of cell-type specific markers for all immune cell types, validating the assignment of various immune cell types with consensus cell types.
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A summary of all the consensus cell types observed in all other ScPCA samples can be found in Figure {@fig:figS6}.
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### Augmenting cell type annotations for malignant cell identification
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Because the consensus annotations were derived from automated methods that do not specifically consider tumor cell states, they provide limited information for distinguishing malignant from normal cells.
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We therefore sought complementary avenues to increase the value of cell type annotations with information that can be leveraged for this purpose.
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In parallel to developing the ScPCA Portal, we launched the OpenScPCA project [@url: https://openscpca.readthedocs.io], an open-science collaborative initiative to further characterize and analyze Portal data.
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Thus far, we have added manual cell type annotations for two projects, `SCPCP000004` (neuroblastoma) and `SCPCP000015` (Ewing sarcoma), to the Portal based on analyses performed in the OpenScPCA project.
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Figure {@fig:fig5}A displays, for example, a UMAP of all libraries in `SCPCP000004` highlighting this project's OpenScPCA annotations which were derived using the `NBAtlas` dataset as reference [@doi:10.1016/j.celrep.2024.114804].
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Unlike the consensus cell type annotations, the OpenScPCA project annotations distinguish between normal and malignant cells and contain far fewer uncharacterized cells.
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Indeed, for `SCPCP000004`, the consensus cell type procedure labeled only ~43% of cells, but the OpenScPCA project labeled ~91% of cells, thereby adding substantial value to the data.
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Note that the Portal's summary cell type report will include comparisons between annotations made in `scpca-nf` to OpenScPCA annotations for relevant libraries.
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In an effort to identify potential malignant cells across all samples in the Portal, we included a step in the `scpca-nf` pipeline to run `InferCNV` [@url:https://github.com/broadinstitute/inferCNV] to quantify copy-number alterations (Figure {@fig:fig2}A).
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The estimates complement the consensus cell types by providing a proxy for a cell's malignant status, such that cells with high levels of CNV are more likely to be tumor than normal cells.
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Indeed, there is broad correspondence between malignant cells (Figure {@fig:fig5}A) and the total per-cell CNV across libraries in `SCPCP000004` (Figure {@fig:fig5}B); malignant cells tend to have higher levels of CNV, whereas normal cells tend to have lower levels of CNV.
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We probed this relationship further within a single neuroblastoma library, `SCPCL000130`, finding clear signatures of canonical neuroblastoma CNV events such as `1q` loss, `11q` gain, and `17p` loss [@doi:10.1038/nrdp.2016.78; @doi:10.1016/j.celrep.2024.114804; @doi:10.1158/2159-8290.CD-14-0622] within malignant cells (Figure {@fig:fig5}C).
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By contrast, normal cells show very few CNV events, consistent with their annotations.
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Most intriguingly, unknown cells show CNV event signatures more similar to the malignant cells than to the normal cells, suggesting many of these cells may indeed be malignant.
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We also see traces of this relationship even when looking at the consensus cell types in conjunction with CNV events.
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In Figure {@fig:fig5}D, we show the distributions of per-cell total CNV events for the most commonly-observed consensus cell types in the neuroblastoma library `SCPCL000130`.
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Here, Unknown and neuron cells have distinctly higher total CNV values compared to other cell types, suggesting that they are likely to be malignant cells.
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We see similar patterns for the ganglioglioma library `SCPCL000049` (Figure {@fig:figS4}B-C), where consensus T cells have low total CNV values, while other cell types including oligodendrocyte precursor cells, neuron associated cells, and Unknown cells have much higher total CNV values.
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As such, joint information from consensus cell type annotations and `InferCNV` results may be used to identify malignant cells across libraries in the Portal, including those which do not yet have associated OpenScPCA project annotations.
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## Analysis of bulk RNA-seq
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Several projects in the ScPCA Portal contain bulk RNA-seq data in addition to single-cell/nuclei RNA-seq data.

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