Anatomy and Neuroscience - Research Publications

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    Sincast: a computational framework to predict cell identities in single-cell transcriptomes using bulk atlases as references
    Deng, Y ; Choi, J ; Cao, K-AL (OXFORD UNIV PRESS, 2022-03-31)
    Characterizing the molecular identity of a cell is an essential step in single-cell RNA sequencing (scRNA-seq) data analysis. Numerous tools exist for predicting cell identity using single-cell reference atlases. However, many challenges remain, including correcting for inherent batch effects between reference and query data andinsufficient phenotype data from the reference. One solution is to project single-cell data onto established bulk reference atlases to leverage their rich phenotype information. Sincast is a computational framework to query scRNA-seq data by projection onto bulk reference atlases. Prior to projection, single-cell data are transformed to be directly comparable to bulk data, either with pseudo-bulk aggregation or graph-based imputation to address sparse single-cell expression profiles. Sincast avoids batch effect correction, and cell identity is predicted along a continuum to highlight new cell states not found in the reference atlas. In several case study scenarios, we show that Sincast projects single cells into the correct biological niches in the expression space of the bulk reference atlas. We demonstrate the effectiveness of our imputation approach that was specifically developed for querying scRNA-seq data based on bulk reference atlases. We show that Sincast is an efficient and powerful tool for single-cell profiling that will facilitate downstream analysis of scRNA-seq data.
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    An integrated analysis of human myeloid cells identifies gaps in in vitro models of in vivo biology
    Rajab, N ; Angel, PW ; Deng, Y ; Gu, J ; Jameson, V ; Kurowska-Stolarska, M ; Milling, S ; Pacheco, CM ; Rutar, M ; Laslett, AL ; Cao, K-AL ; Choi, J ; Wells, CA (CELL PRESS, 2021-06-08)
    The Stemformatics myeloid atlas is an integrated transcriptome atlas of human macrophages and dendritic cells that systematically compares freshly isolated tissue-resident, cultured, and pluripotent stem cell-derived myeloid cells. Three classes of tissue-resident macrophage were identified: Kupffer cells and microglia; monocyte-associated; and tumor-associated macrophages. Culture had a major impact on all primary cell phenotypes. Pluripotent stem cell-derived macrophages were characterized by atypical expression of collagen and a highly efferocytotic phenotype. Myeloid subsets, and phenotypes associated with derivation, were reproducible across experimental series including data projected from single-cell studies, demonstrating that the atlas provides a robust reference for myeloid phenotypes. Implementation in Stemformatics.org allows users to visualize patterns of sample grouping or gene expression for user-selected conditions and supports temporary upload of your own microarray or RNA sequencing samples, including single-cell data, to benchmark against the atlas.
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    HBO1 is required for the maintenance of leukaemia stem cells
    MacPherson, L ; Anokye, J ; Yeung, MM ; Lam, EYN ; Chan, Y-C ; Weng, C-F ; Yeh, P ; Knezevic, K ; Butler, MS ; Hoegl, A ; Chan, K-L ; Burr, ML ; Gearing, LJ ; Willson, T ; Liu, J ; Choi, J ; Yang, Y ; Bilardi, RA ; Falk, H ; Nghi, N ; Stupple, PA ; Peat, TS ; Zhang, M ; de Silva, M ; Carrasco-Pozo, C ; Avery, VM ; Khoo, PS ; Dolezal, O ; Dennis, ML ; Nuttall, S ; Surjadi, R ; Newman, J ; Ren, B ; Leaver, DJ ; Sun, Y ; Baell, JB ; Dovey, O ; Vassiliou, GS ; Grebien, F ; Dawson, S-J ; Street, IP ; Monahan, BJ ; Burns, CJ ; Choudhary, C ; Blewitt, ME ; Voss, AK ; Thomas, T ; Dawson, MA (NATURE PORTFOLIO, 2020-01-09)
    Acute myeloid leukaemia (AML) is a heterogeneous disease characterized by transcriptional dysregulation that results in a block in differentiation and increased malignant self-renewal. Various epigenetic therapies aimed at reversing these hallmarks of AML have progressed into clinical trials, but most show only modest efficacy owing to an inability to effectively eradicate leukaemia stem cells (LSCs)1. Here, to specifically identify novel dependencies in LSCs, we screened a bespoke library of small hairpin RNAs that target chromatin regulators in a unique ex vivo mouse model of LSCs. We identify the MYST acetyltransferase HBO1 (also known as KAT7 or MYST2) and several known members of the HBO1 protein complex as critical regulators of LSC maintenance. Using CRISPR domain screening and quantitative mass spectrometry, we identified the histone acetyltransferase domain of HBO1 as being essential in the acetylation of histone H3 at K14. H3 acetylated at K14 (H3K14ac) facilitates the processivity of RNA polymerase II to maintain the high expression of key genes (including Hoxa9 and Hoxa10) that help to sustain the functional properties of LSCs. To leverage this dependency therapeutically, we developed a highly potent small-molecule inhibitor of HBO1 and demonstrate its mode of activity as a competitive analogue of acetyl-CoA. Inhibition of HBO1 phenocopied our genetic data and showed efficacy in a broad range of human cell lines and primary AML cells from patients. These biological, structural and chemical insights into a therapeutic target in AML will enable the clinical translation of these findings.
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    A simple, scalable approach to building a cross-platform transcriptome atlas
    Angel, PW ; Rajab, N ; Deng, Y ; Pacheco, CM ; Chen, T ; Le Cao, K-A ; Choi, J ; Wells, CA ; Fertig, EJ (PUBLIC LIBRARY SCIENCE, 2020-09-01)
    Gene expression atlases have transformed our understanding of the development, composition and function of human tissues. New technologies promise improved cellular or molecular resolution, and have led to the identification of new cell types, or better defined cell states. But as new technologies emerge, information derived on old platforms becomes obsolete. We demonstrate that it is possible to combine a large number of different profiling experiments summarised from dozens of laboratories and representing hundreds of donors, to create an integrated molecular map of human tissue. As an example, we combine 850 samples from 38 platforms to build an integrated atlas of human blood cells. We achieve robust and unbiased cell type clustering using a variance partitioning method, selecting genes with low platform bias relative to biological variation. Other than an initial rescaling, no other transformation to the primary data is applied through batch correction or renormalisation. Additional data, including single-cell datasets, can be projected for comparison, classification and annotation. The resulting atlas provides a multi-scaled approach to visualise and analyse the relationships between sets of genes and blood cell lineages, including the maturation and activation of leukocytes in vivo and in vitro. In allowing for data integration across hundreds of studies, we address a key reproduciblity challenge which is faced by any new technology. This allows us to draw on the deep phenotypes and functional annotations that accompany traditional profiling methods, and provide important context to the high cellular resolution of single cell profiling. Here, we have implemented the blood atlas in the open access Stemformatics.org platform, drawing on its extensive collection of curated transcriptome data. The method is simple, scalable and amenable for rapid deployment in other biological systems or computational workflows.