1. Define context(s)
reveal new biological insights
Primary goal of the model/tool/database
Single-cell RNA-sequencing (scRNA-seq) offers unprecedented resolution for studying cellular decision-making processes. Robust inference of cell state transition paths and probabilities is an important yet challenging step in the analysis of these data. Here we present scEpath, an algorithm that calculates energy landscapes and probabilistic directed graphs in order to reconstruct developmental trajectories. We quantify the energy landscape using ‘single-cell energy’ and distance-based measures, and find that the combination of these enables robust inference of the transition probabilities and lineage relationships between cell states. We also identify marker genes and gene expression patterns associated with cell state transitions. Our approach produces pseudotemporal orderings that are—in combination—more robust and accurate than current methods, and offers higher resolution dynamics of the cell state transitions, leading to new insight into key transition events during differentiation and development. Moreover, scEpath is robust to variation in the size of the input gene set, and is broadly unsupervised, requiring few parameters to be set by the user.
Biological domain of the model
scRNA-seq data of various tissues
Structure(s) of interest in the model
celluar trajectories, cell state landscape
Spatial scales included in the model
cells to tissues
Time scales included in the model
seconds to weeks
2. Data for building and validating the model
Data for building the model |
Published? |
Private? |
How is credibility checked? |
Current Conformance Level / Target Conformance Level |
in vitro (primary cells cell, lines, etc.) |
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ex vivo (excised tissues) |
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in vivo pre-clinical (lower-level organism or small animal) |
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in vivo pre-clinical (large animal) |
Yes |
No |
The model was built in an unsupervised way on unbiased single-cell RNA sequencing data. |
Extensive |
Human subjects/clinical |
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Other: ________________________ |
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Data for validating the model |
Published? |
Private? |
How is credibility checked? |
Current Conformance Level / Target Conformance Level |
in vitro (primary cells cell, lines, etc.) |
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ex vivo (excised tissues) |
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in vivo pre-clinical (lower-level organism or small animal) |
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in vivo pre-clinical (large animal) |
Yes |
No |
By 1) comparing the pseudotime derived by the model to known temporal sequence and 2) by comparing the developmental events identified by the model to known developmental branches. |
Adequate |
Human subjects/clinical |
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Other: ________________________ |
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3. Validate within context(s)
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Who does it? |
When does it happen? |
How is it done? |
Current Conformance Level / Target Conformance Level |
Verification |
Students/postdocs/investigators |
Throughout the project |
1) The generated cell state landscape agrees with known overall structure. 2) The pseudotime ordering agrees well with known temporal sequence of cell states. |
Extensive |
Validation |
Students/postdocs/investigators |
As the unsupervised model was established |
1) The inferred cell state landscape and the cell state transition are validated by known cell types and their developmental relationships. 2) The pseudotime ordering is validated by known temporal ordering of cell types and known key regulatory events are recovered |
Extensive |
Uncertainty quantification |
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Sensitivity analysis |
Students/postdocs/investigators |
As the unsupervised model was established |
By tuning key parameters and comparing to annotated data. |
Adequate |
Other:__________ |
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Additional Comments |
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4. Limitations
Disclaimer statement (explain key limitations) |
Who needs to know about this disclaimer? |
How is this disclaimer shared with that audience? |
Current Conformance Level / Target Conformance Level |
The technical noise in single-cell RNA sequencing data might cause inaccuracy and sufficient number of cells might by needed for accurate energy landscape estimation. |
Scientific community who intends to apply this method to raw scRNA-seq data. |
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Adequate |
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5. Version control
Current Conformance Level / Target Conformance Level |
Extensive |
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Naming Conventions? |
Repository? |
Code Review? |
individual modeler |
Yes |
Yes |
peers |
within the lab |
Yes |
Yes |
peers |
collaborators |
Yes |
Yes |
via regular meetings |
6. Documentation
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Current Conformance Level / Target Conformance Level |
Code commented? |
Adequate |
Scope and intended use described? |
Extensive |
User’s guide? |
Extensive |
Developer’s guide? |
Partial |
7. Dissemination
Current Conformance Level / Target Conformance Level |
Extensive |
Target Audience(s): |
“Inner circle” |
Scientific community |
Public |
Simulations |
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Models |
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Software |
package: https://github.com/sqjin/scEpath |
package: https://github.com/sqjin/scEpath |
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Results |
Shared folders |
Paper and tutorials |
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Implications of results |
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8. Independent reviews
Current Conformance Level / Target Conformance Level |
To be done |
Reviewer(s) name & affiliation: |
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When was review performed? |
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How was review performed and outcomes of the review? |
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9. Test competing implementations
Current Conformance Level / Target Conformance Level |
Adequate |
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Yes or No (briefly summarize) |
Were competing implementations tested? |
Yes. The method has been compared to several other commonly used methods on benchmark datasets. |
Did this lead to model refinement or improvement? |
Yes. |