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Bosotti, R.

Publications and source records attributed to Bosotti, R..

3 recordsLinked to original sources

COG5-mediated Golgi organization coordinates neural progenitor fate during human brain development

Apical radial glial cells (aRGs) are highly polarized epithelial cells that drive human cortical development through tightly regulated lineage progression. A distinctive feature of aRGs is their non-canonical Golgi organization, raising the possibility that specialized intracellular trafficking and secretory processes contribute to progenitor behaviour. However, how Golgi architecture contributes to human corticogenesis remains poorly understood. Here, we investigate this question in the context of COG5 deficiency, a rare congenital disorder of glycosylation associated with neurodevelopmental defects, using patient-derived fibroblasts and brain organoids, N-glycomics, glycolipidomics, and single-cell transcriptomics. We show that COG5 deficiency disrupts Golgi organization and impairs Golgi-to-ER retrograde trafficking. At the neurodevelopmental level, COG5-deficient organoids display altered ventricular organization and reduced basal progenitor abundance. Single-cell transcriptomic analysis reveals rewired developmental trajectories characterized by premature differentiation from apical progenitors with activation of neuronal pathways. Despite the absence of major abnormalities in plasma glycoproteins glycosylation, glycolipidomic analysis identified marked defects in glycosphingolipid maturation within patient-derived brain organoids, indicating selective impairment of neural lipid glycosylation. Transcriptomic and biochemical analyses further revealed dysregulation of developmental pathways, including reduced Notch signalling and altered localization of both cleaved Notch1 and the Golgi-associated protein ACBD3. Consistent with impaired progenitor maintenance, COG5-deficient organoids displayed altered cleavage plane orientation in apical progenitors. Together, our findings suggest a link between Golgi organization, intracellular trafficking, membrane glycosylation, and lineage progression in human neural progenitors, identifying Golgi architecture as an important regulator of apical radial glial cell biology and cortical development.

cell biology↗

Human basal radial glia morphotypes are transcriptionally distinct and exhibit different cell fate determination

Basal radial glia (bRG) are key neural progenitors driving human neocortical expansion. They exhibit remarkable morphological heterogeneity, yet the stability and functional significance of their distinct morphotypes remains unclear. Using human cortical brain organoids combined with long-term live imaging and morphology-resolved spatial transcriptomics (CellShape-seq), we show that bRG morphotypes display distinct morphodynamic behaviors, proliferative capacities and transcriptional profiles. While bifurcated bRG remodel extensively during mitosis to produce morphologically diverse progeny, multipolar cells are most morphologically flexible during interphase. Multipolar bRG further show the greatest proliferative capacity and the transcriptional signature related to progenitor state. Bifurcated bRG are least proliferative and are enriched for the multifunctional gene expression regulator YBX1. Pharmacological inhibition of YBX1 depletes bifurcated bRG, reduces neurogenesis, and promotes glial commitment. Our findings link progenitor morphology, gene expression and fate, providing a framework for understanding the cellular logic of human cortical development.

neuroscience↗

Molecular phenotyping of breast cancer cell lines at the single-cell level for automated cancer diagnosis and prediction of drug sensitivity

Breast cancer patient stratification is mainly driven by tumour receptor status and histological grading and subtyping, with about twenty percent of patients for which absence of any actionable biomarkers results in no clear therapeutic intervention. Cancer cells within the same tumour have heterogeneous phenotypes and exhibit dynamic plasticity. However, how to evaluate such heterogeneity and its impact on outcome and drug response is still unclear. Here, we transcriptionally profiled 35,276 individual cells from 32 breast cancer cell lines covering all main breast cancer subtypes to yield a breast cancer cell line atlas. We found high degree of heterogeneity in the expression of clinically relevant biomarkers across individual cells within the same cell line; such heterogeneity is non-genetic and dynamic. We computationally mapped single cell transcriptional profiles of patients tumour biopsies to the atlas to determine their composition in terms of cell lines. Each tumour was found to be heterogenous and composed of multiple cell lines mostly, but not exclusively, of the same subtype. We then trained an algorithm on the atlas to determine cell line composition from bulk gene expression profiles of tumour biopsies, thus providing a novel approach to patient stratification. Finally, we linked results from large-scale in vitro drug screening1,2 to the single cell data to computationally predict responses to more than 450 anticancer agents starting from single-cell transcriptional profiles. We thus found that transcriptional heterogeneity enables cells with differential drug sensitivity to co-exist in the same population. Our work provides a unique resource and a novel framework to determine tumour heterogeneity and drug response in breast cancer patients.

cancer biology↗