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Erlandsson, E.

Publications and source records attributed to Erlandsson, E..

5 recordsLinked to original sources

Scarf: A toolkit for memory efficient analysis of large-scale single-cell genomics data

The increasing capacity to perform large-scale single-cell genomic experiments continues to outpace the computational requirements to efficiently handle growing datasets. Herein we present Scarf, a modularly designed Python package that seamlessly interoperates with other single-cell toolkits and allows for memory-efficient single-cell analysis of millions of cells on a laptop or low-cost devices like single board computers. We demonstrate Scarfs memory and compute-time efficiency by applying it to the largest existing single-cell RNA-Seq and ATAC-Seq datasets. Scarf wraps memory-efficient implementations of a graph-based t-stochastic neighbour embedding and hierarchical clustering algorithm. Moreover, Scarf performs accurate reference-anchored mapping of datasets while maintaining memory efficiency. By implementing a novel data downsampling algorithm, Scarf additionally can generate representative sampling of cells from a given dataset wherein rare cell populations and lineage differentiation trajectories are conserved. Together, Scarf provides a framework wherein any researcher can perform advanced processing, downsampling, reanalysis, and integration of atlas-scale datasets on standard laptop computers.

bioinformatics↗

Single-Cell Multiomics Reveals Distinct Cell States at the Top of Human Hematopoietic Hierarchy

The advent of single cell (Sc) genomics has challenged the dogma of haematopoiesis as a tree-like structure of stepwise lineage commitment through distinct and increasingly restricted progenitor populations. Instead, analysis of ScRNA-seq has proposed that the earliest events in human hematopoietic stem cell (HSC) differentiation are characterized by only subtle molecular changes, with hematopoietic stem and progenitor cells (HSPCs) existing as a continuum of low-primed cell-states that gradually transition into a specific lineage (CLOUD-HSPCs). Here, we combine ScRNA-seq, ScATAC-seq and cell surface proteomics to dissect the heterogeneity of CLOUD-HSPCs at different stages of human life. Within CLOUD-HSPCs, pseudotime ordering of both mRNA and chromatin data revealed a bifurcation of megakaryocyte/erythroid and lympho/myeloid trajectories immediately downstream a subpopulation with an HSC-specific enhancer signature. Importantly, both HSCs and lineage-restricted progenitor populations could be prospectively isolated based on correlation of their molecular signatures with CD35 and CD11A expression, respectively. Moreover, we describe the changes that occur in this heterogeneity as hematopoiesis develops from neonatal to aged bone marrow, including an increase of HSCs and depletion of lympho-myeloid biased MPPs. Thus, this study dissects the heterogeneity of human CLOUD-HSPCs revealing distinct HSPC-states of relevance in homeostatic settings such as ageing.

cell biology↗

Nabo - a framework to define leukemia-initiating cells and differentiation in single-cell RNA-sequencing data

Single-cell transcriptomics facilitates innovative approaches to define and identify cell types within tissues and cell populations. An emerging interest in the cancer field is to assess the heterogeneity of transformed cells, including the identification of tumor-initiating cells based on similarities to their normal counterparts. However, such cell mapping is often confounded by the large effects on total gene expression programs introduced by strong perturbations such as an oncogenic event. Here, we present Nabo, a novel computational method that allows mapping of cells from one population to the most similar cells in a reference population, independently of confounding changes to gene expression programs initiated by perturbation. We validated this method on multiple datasets from different sources and platforms and show that Nabo achieves higher rates of accuracy than conventional classification methods. Nabo is available as an integrated toolkit for preprocessing, cell mapping, differential gene expression identification, and visualization of single-cell RNA-Seq data. For exploratory studies, Nabo includes methods to help evaluate the reliability of cell mapping results. We applied Nabo on droplet-based single-cell RNA-Seq data of healthy and oncogene-induced (MLL-ENL) hematopoietic progenitor cells (GMLPs) differentiating in vitro. Despite a substantial cellular heterogeneity resulting from differentiation of GMLPs and the large transcriptional effects induced by the fusion oncogene, Nabo could pinpoint the specific cell stage where differentiation arrest occurs, which included an immunophenotypic definition of the tumor-initiating population. Thus, Nabo allows for relevant comparison between target and control cells, without being confounded by differences in population heterogeneity.

bioinformatics↗

Concurrent stem- and lineage-affiliated chromatin programs precede hematopoietic lineage restriction

The emerging notion of hematopoietic stem- and progenitor cells (HSPCs) as a low-primed cloud without sharply demarcated gene expression programs raises the question on how cellular fate options emerge, and at which stem-like stage lineage priming is initiated. Here we investigated single-cell chromatin accessibility of Lineage-, cKit+, Sca1+ (LSK) HSPCs spanning the early differentiation landscape. Application of a signal-processing algorithm to detect transition points corresponding to massive alterations in accessibility of 571 transcription factor-motifs revealed a population of LSK FMS-like tyrosine kinase 3(Flt3)intCD9high cells that concurrently display stem-like and lineage-affiliated chromatin signatures pointing to a simultaneous gain of both Lympho-Myeloid and Megakaryocyte-Erythroid programs. Molecularly and functionally, these cells position between stem cells and committed progenitors, display multi-lineage capacity in vitro and in vivo, but lack self-renewal activity. This integrative molecular analysis resolves the heterogeneity of cells along hematopoietic differentiation and permits investigation of chromatin-mediated transition between multipotency and lineage restriction.

cell biology↗

HSCs contribute actively to native multilineage hematopoiesis but with reduced differentiation capacity upon aging

A hallmark of adult hematopoiesis is the continuous replacement of blood cells with limited lifespans. It is well established that adult hematopoietic stem cells (HSCs) are active contributors to these processes after transplantation, yet their role in native hematopoiesis has recently been called into question. Here, we use inducible lineage tracing from genetically marked adult HSCs to explore their roles in the steady state. We show that adult HSCs contribute robustly to all lineages via intermediate progenitor cells, but with neglible production of hematopoietic cells with a known fetal origin. We further reveal that the timing for regeneration of distinct blood lineages varies substantially. Finally, HSC contribution to multilineage hematopoiesis in aged animals declines with increasing age. Therefore, while HSCs are active contributors to native adult hematopoiesis, it appears that the numerical increase of HSCs is a physiologically relevant compensatory mechanism to account for a reduced differentiation capacity with age.

developmental biology↗