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Standardizing workflows in imaging transcriptomics with the abagen toolbox

  1. Ross D Markello  Is a corresponding author
  2. Aurina Arnatkeviciute
  3. Jean-Baptiste Poline
  4. Ben D Fulcher
  5. Alex Fornito
  6. Bratislav Misic  Is a corresponding author
  1. McConnell Brain Imaging Centre, Montreal Neurological Institute, McGill University, Canada
  2. School of Psychological Sciences & Monash Biomedical Imaging, Monash University, Australia
  3. School of Physics, University of Sydney, Australia
Tools and Resources
Cite this article as: eLife 2021;10:e72129 doi: 10.7554/eLife.72129
5 figures, 1 table and 2 additional files

Figures

Processing choices influence transcriptomic analyses.

(a) Examples of the three analyses used to assess differences in gene expression matrices generated by transcriptomic pipelines. First row: a depiction of the region-by-gene expression matrix generated from one of the 746,496 tested processing pipelines. Second row, left: we compute the correlation between rows of each matrix to generate a symmetric region × region CGE matrix. We then compute the correlation between the upper triangle of this CGE matrix and the upper triangle of a regional distance matrix to examine the degree to which CGE decays with increasing distance between regions (Arnatkeviciute et al., 2019). Second row, middle: we compute the Euclidean distance between columns of each matrix to generate a gene × gene GCE matrix. We use previously defined functional gene communities (Oldham et al., 2008) to compute a silhouette score for this GCE matrix to investigate whether genes within a module have more similar patterns of spatial expression than genes between modules. Second row, right: the first principal component is extracted from the RGE matrix. We compute the correlation between this principal component and the whole-brain T1w/T2w ratio (Burt et al., 2018) to understand how closely these maps covary across the brain. (b) The full statistical distributions from each of the three analyses for all 746,496 pipelines. Left panel: Spearman correlation values, ρ, from the CGE analyses. Middle panel: silhouette scores from the GCE analyses. Right panel: Spearman correlation coefficients, ρ, from the RGE analyses. CGE: correlated gene expression; GCE: gene co-expression; RGE: regional gene expression.

Parameter choice differentially impacts statistical estimates.

(a) Rank of the relative importance for each parameter (y-axis) across all three analyses (x-axis). Warmer colors indicate parameters that have a greater influence on statistical estimates. (b) Statistical distributions from the three analyses, shown as kernel density plots, separated by choice of gene normalization method (the most impactful parameter as shown in panel a). (c) Density plots of the statistical estimates for all 746,496 pipelines shown along the first two principal components, derived from the 746,496 (pipeline) x 3 (statistical estimates) matrix, representing how different the statistical estimates from each of the three analyses are relative to other pipelines. Left panel: pipelines are colored based on choice of gene normalization method, where each color represents 1/3 of the pipelines. Here, the pipelines in which no normalization was applied (purple) are distinguished from those in which some form of normalization was applied (blue and brown). Right panel: pipelines are colored based on whether gene normalization was performed within (True, red) or across (False, purple) structural classes (i.e. cortex, subcortex/brainstem, cerebellum; see Materials and methods: Gene expression pipelines for more information).

Reproducing published pipelines.

(a) Parameter choices used in the reproduction of published pipelines. Processing steps with categorical choices (e.g., gene normalization) were converted to numerical choices for display purposes only. These choices reflect the range of choices enumerated in Table 1. (b) Relative expression values of cortical somatostatin (SST) generated by each of the reproduced pipelines. Value ranges vary based on pipeline processing options. (c) The Pearson correlation between the cortical somatostatin (SST) maps generated by the nine pipelines shown in panel (b). (d) Statistical estimates from the three analyses described in Materials and methods: Analytic approaches applied to expression data from each of the published pipelines.

Workflows and features in the abagen toolbox.

(a) The primary workflow of abagen, used in the reported analyses, accepts a brain atlas and returns a parcellated brain-region-by-gene expression matrix. (b) An alternative abagen workflow accepts a regional mask and returns a processed tissue-sample-by-gene expression matrix, for all tissue samples from the six AHBA donors that fall within boundaries of the mask. (c) Examples of selected features from the abagen workflows and additional toolbox functionality. Top left: examples of some commonly-used atlases that can be employed with the parcellation workflow shown in panel (a). Bottom left: abagen can accept either standard atlases (i.e. in MNI space) or atlases defined in the space of the six individual donors from the AHBA. Top right: an additional workflow available in abagen can be used to generate densely-interpolated expression maps from AHBA data using a k-nearest neighbors interpolation algorithm. Bottom right: using high-resolution atlases in the parcellation workflow (panel a) may result in some parcels being assigned no expression data; abagen supports two methods for assigning values to such regions.

Annotated example abagen report.

Example of an automatically generated methods section report from the abagen toolbox. Processing steps are shown on the left and the relevant methods text—which is updated when these steps are modified—is shown in the same font color on the right. Reports also include a formatted reference section and relevant equations; these are not shown here for conciseness. Note that some processing steps (e.g. normalizing within structures, missing data handling) are omitted here because they are not run by default (see Supplementary file 1).

Tables

Table 1
Abagen pipeline options.

Overview of 17 options to be considered when processing the AHBA data. The Choices column indicates the number of parameters explored in the current report (numerator) and the total number of parameters possible for the given option (denominator). A denominator of n indicates a hypothetically near-infinite parameter space. The Description column gives a brief overview of the processing choice; for more detail refer to the relevant section in Materials and methods: Gene expression pipelines.

OptionChoicesDescription
Volumetric or surface atlas2/2Whether to use a volumetric or surface representation of the atlas
Individualized or group atlas1/2Whether to use individualized donor-specific atlases or a group-level atlas
Use non-linear MNI coordinates2/2Whether to use updated MNI coordinates provided by alleninf package
Mirror samples across L/R hemisphere3/4Whether to mirror (i.e., duplicate) samples across hemisphere boundary
Update probe-to-gene annotations2/2Whether to update probe annotations
Intensity-based filtering threshold3/nThreshold for intensity-based filtering of probes
Inter-areal similarity threshold1/nThreshold for removing samples with low inter-areal correspondence
Probe selection method6/8Method by which to select which probe(s) should represent a given gene
Donor-specific probe selection3/3How specified probe selection should integrate data from different donors
Missing data method2/3How to handle when brain regions are not assigned expression data
Sample-to-region matching tolerance3/nDistance tolerance for matching tissue samples to atlas brain regions
Sample normalization method3/10Method for normalizing tissue samples (across genes)
Gene normalization method3/10Method for normalizing genes (across tissue samples)
Normalize only matched samples2/2Whether to perform gene normalization for all versus matched samples
Normalizing discrete structures2/2Whether to perform gene normalization within structural classes
Sample-to-region combination method2/2Whether to aggregate tissue samples in regions within or across donors
Sample-to-region combination metric2/2Metric for aggregating tissue samples into atlas brain regions

Additional files

Transparent reporting form
https://cdn.elifesciences.org/articles/72129/elife-72129-transrepform1-v2.pdf
Supplementary file 1

Default abagen pipeline options.

The default settings for the 17 processing steps considered when processing the AHBA data with abagen. An entry of ‘—' indicates that this is a required, user-supplied parameter. A blank entry indicates that the processing step is not implemented by default. Refer to Table 1 and Methods: Gene expression pipelines for further details.

https://cdn.elifesciences.org/articles/72129/elife-72129-supp1-v2.pdf

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