Biology subjects

Gospodinova, K. O.

Publications and source records attributed to Gospodinova, K. O..

3 recordsLinked to original sources

bioRxiv2025

SHIP1 regulates TREM2 signalling and macrophage functions in a hiPSC-derived model

Genetic and functional studies strongly implicate microglia in the pathology of Alzheimers disease (AD). The Triggering Receptor Expressed on Myeloid cells 2 (TREM2) pathway is an important functional regulator of microglia in AD, promoting phagocytosis of apoptotic cells, debris and pathogenic proteins including amyloid beta (A{beta}). Additionally, genome-wide association studies have identified risk bearing polymorphisms in several members of the pathway including INPP5D, encoding the inositol phosphatase SHIP1. Recent studies utilising in vitro macrophage and microglia models and preclinical mouse models have identified a role for SHIP1 in both modulating TREM2 signalling and modulating neurotoxic microglial activation. In this study, we characterised the role of SHIP1 in regulating TREM2 signalling and global microglial functions using human induced pluripotent stem cell (hiPSC)-derived macrophages as a physiologically-relevant in vitro model of microglia. Isogenic parent and SHIP1 knockout (SHIP1 KO) lines were generated and macrophage phenotype and functions investigated. Knocking out SHIP1 was associated with a decreased expression of the lipopolysaccharide (LPS) co-receptor CD14, which translated to lower secretion of pro-inflammatory cytokines on stimulation with LPS. Additionally, SHIP1 KO hiPSC-derived macrophages showed decreased signalling upon TREM2 stimulation, which was further reflected in a reduced level of phagocytosis of apoptotic neurons compared to the parental line. The observed attenuated TREM2 signalling did not correspond to decreased TREM2 protein levels nor increased shedding of the receptor. Examination of macrophage scavenger receptor expression revealed reduced cell surface levels of CD163 and CD206 involved in resolution of inflammation but increased levels of the phagocytic receptor MerTK, while morphological analysis revealed a less amoeboid activated phenotype and increased adhesion. Taken together, our data indicates that SHIP1 plays a key role in regulating global macrophage functions in addition to modulating TREM2 signalling and inflammation.

immunology↗

bioRxiv2023

Morphological profiling by cell painting in human neural progenitor cells classifies hit compounds in a pilot drug screen for Alzheimer's disease

1Alzheimers disease (AD) accounts for 60-70% of dementia cases. Current treatments are inadequate and there is a need to develop new approaches to AD drug discovery. We chose to develop a cell phenotype-based drug screen centred on the AD-risk gene, SORL1, which encodes the protein SORLA. Increased AD risk has been repeatedly linked to variants in SORL1, particularly those that confer loss of, or decreased, SORLA. This is consistent with the lower SORL1 levels observed in post-mortem brain samples from individuals with AD. Consistent with its role in the endolysosomal pathway, deletion of SORL1 is associated with enlarged endosomes in neural progenitor cells (NPCs) and neurons. We, therefore, hypothesised that multiparametric, image-based phenotyping would identify features characteristic of SORL1 deletion. An automated morphological profiling assay (known as Cell Painting) was adapted to wild-type and SORL1-/- NPCs. This methodology was used to determine the phenotypic response of SORL1-/- NPCs to treatment with compounds from a small FDA/internationally-approved drug library (TargetMol, 330 compounds). We detected distinct phenotypic signatures for SORL1-/- NPCs compared to isogenic wild-type controls. Furthermore, we identified 16 approved drugs that reversed the mutant morphological signatures in NPCs derived from 3 SORL1-/- subclonal iPSC lines. Network pharmacology analysis revealed the 16 compounds belonged to five mechanistic groups: 20S proteasome, aldehyde dehydrogenase, topoisomerase I and II, and DNA synthesis inhibitors. Enrichment analysis confirmed targeting to gene sets associated with these annotated targets, and to pathways/biological processes associated with DNA synthesis/damage/repair, Proteases/proteasome and metabolism._Prediction of novel targets for some compounds revealed enrichment in pathways associated with neural cell function and AD. The findings suggest that image-based phenotyping by morphological profiling distinguishes SORL1-/- NPCs from isogenic wild-type lines, and predicts treatment responses that rescue SORL1-/--associated cellular signatures that are relevant to both SORLA function and AD. Overall, this work suggests that i) a quantitative phenotypic metric can distinguish iPSC-derived SORL1-/- NPCs from isogenic wild-type control and ii) phenotypic screening combined with multiparametric high-content image analysis is a viable option for drug repurposing and discovery in this human neural cell model of Alzheimers disease.

neuroscience↗

bioRxiv2021

Loss of SORCS2 is associated with neuronal DNA double-strand breaks

SORCS2 is one of five proteins that constitute the Vps10p-domain receptor family. Members of this family play important roles in cellular processes linked to neuronal survival, differentiation and function. Genetic and functional studies implicate SORCS2 in cognitive function, as well as in neurodegenerative and psychiatric disorders. DNA damage and DNA repair deficits are linked to ageing and neurodegeneration, and transient neuronal DNA double-strand breaks (DSBs) also occur as a result of neuronal activity. Here, we report a novel role for SORCS2 in DSB formation. We show that SorCS2 loss is associated with elevated DSB levels in the mouse dentate gyrus and that knocking out SORCS2 in a human neuronal cell line increased Topoisomerase II{beta}-dependent DSB formation and reduced neuronal viability. Neuronal stimulation had no impact on levels of DNA breaks in vitro, suggesting that the observed differences may not be the result of aberrant neuronal activity in these cells. Our findings are consistent with studies linking the VPS10 receptors and DNA damage to neurodegenerative conditions.

neuroscience↗