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Biology subjects

de Almeida, G. P.

Publications and source records attributed to de Almeida, G. P..

3 recordsLinked to original sources

Concurrent de novo inference of cell types and gene regulatory networks with NetMap

Existing methods for GRN inference based on single-cell RNA-sequencing (scRNA-seq) data typically require prior cell type calling and then infer GRNs at cell type resolution. This risks propagation of errors from cell annotation to GRN inference. Moreover, it makes existing methods blind to regulatory programs that are shared between different cell types, as well as to subtle differences in regulatory activity between cells of the same type. To address these limitations, we present NetMap, a pipeline that jointly infers cell-level GRNs and clusters cells based on shared regulatory programs, combining state-of-the-art single-cell autoencoders and explainable artificial intelligence methods. NetMap outperforms existing methods on synthetic data with a known ground-truth GRN and is able to reliably detect biologically plausible regulons on real data. Moreover, requiring only a small set of user-provided marker genes, NetMap can be used for GRN structure-informed probabilistic cell type assignment. We demonstrate the utility of this approach using PBMC data, where we achieve near perfect clustering and cell type recovery performance compared with manually created cell type labels. Finally, we applied NetMap to a data set with naive, activated and exhausted T-cells, and were able to verify that NetMap retrieves relevant T-cell state signatures from the data, with two approaches. NetMap can be installed as a python package from https://github.com/bionetslab/netmap and is compatible with the scverse system. Scripts to reproduce our results can be found at https://github.com/bionetslab/netmap-evaluation.

bioinformatics↗

Morc3 silences endogenous retroviruses by enabling Daxx-mediated H3.3 incorporation

Endogenous retroviruses (ERVs) comprise a significant portion of mammalian genomes. Although, specific ERV loci feature regulatory roles for host gene expression, most ERV integrations are transcriptionally repressed by Setdb1 mediated H3K9me3 and DNA methylation. However, the protein network which regulates deposition of these chromatin modifications is still incompletely understood. Here, we performed a genome-wide sgRNA screen for genes involved in ERV silencing and identified the GHKL ATPase protein Morc3 as top scoring hit. Morc3 knock-out cells display de-repression, reduced H3K9me3 and increased chromatin accessibility of distinct ERV classes. We found that the GHKL ATPase domain of Morc3 is critical for ERV silencing, since mutants which cannot bind ATP, or which are defective in ATP hydrolysis cannot rescue the Morc3 ko phenotype. Proteomic analysis revealed that Morc3 mutant protein which cannot bind ATP fails to interact with the H3.3 chaperone Daxx. This interaction depends on Morc3 SUMOylation, as Daxx lacking the SUMO interaction domain shows reduced association with Morc3. Notably, in Morc3 ko cells, we observed strongly reduced H3.3 on Morc3 binding sites. Thus, our data demonstrate Morc3 as critical regulator of Daxx-mediated H3.3 incorporation to ERV regions.

molecular biology↗

Transcriptional profiling and single-cell chimerism analysis identifies human tissue resident T cells in the human skin after allogeneic stem cell transplantation

Tissue resident memory T cells (TRM) have recently emerged as crucial cellular players for host defense in a wide variety of tissues and barrier sites. Mouse models revealed that they are maintained long-term in loco unlike recirculating effector memory T cells (TEM). Insights into the maintenance and regulatory checkpoints of human tissue resident T cells (TRM) remain scarce, especially due to the obstacles in tracking T cells over time and system-wide in humans. We present a clinical model that allowed us to overcome these limitations. We demonstrate that allogeneic stem cell transplantation resulted in compartmentalization of host T cells in the human skin despite complete donor T cell chimerism in the blood, thus unmasking long-term persistence of tissue resident T cell subsets of host origin within the diverse skin T cell community. Single-cell transcriptional profiling paired with single-cell chimerism analysis provided an in-depth characterization of these bona fide skin resident T cells. Their phenotype, functions and regulatory checkpoints may serve therapeutic strategies for the treatment of autoimmune diseases and chronic infections, where their specific depletion versus maintenance, respectively, will have to be harnessed pharmaceutically.

immunology↗