Biology subjects

Cerisoli, M.

Publications and source records attributed to Cerisoli, M..

3 recordsLinked to original sources

bioRxiv2025

aDISCO: A Broadly Applicable Method for 3D Microscopy of Archival Paraffin-Embedded Human Tissues

Human surgery and autopsy specimens are routinely stored as formalin-fixed paraffin-embedded (FFPE) tissue blocks. Diagnoses are based on microscopic examination of two-dimensional sections. Good clinical practice requires that samples be retained for decades, thus giving rise to vast archives of healthy and diseased tissues. While tissue clearing and whole-mount microscopy enable 3D analysis, FFPE human tissue blocks are often unsuitable for clearing and immunolabeling due to their large size, extensive cross-linking, and wax embedding. Here, we introduce archival DISCO (aDISCO), a versatile and robust clearing method designed to overcome these challenges. aDISCO achieves complete clearing and immunolabeling of large samples stored for 15 years or more. We applied aDISCO to human brain, spinal cord, peripheral nerve, skin, muscle, heart, kidney, liver, spleen, colon, and lung, using a broad range of antibodies. To investigate the clinical usefulness of quantitative 3D histology by aDISCO, we performed a case study of focal cortical dysplasia, a neurodevelopmental disease causing epilepsy. Using deep-learning-based analysis, we found disrupted cortical layering and both focal and global neuronal density variations in FCD, features that are likely to be overlooked by conventional histology. In summary, aDISCO delivers large datasets suitable for deep-learning-based processing, enabling the detection of subtle and sparse pathologies in large archival human tissue specimens.

bioengineering↗

bioRxiv2025

Gpnmb Defines a Phagocytic State of Microglia Linked to Neuronal Loss in Prion Disease

Neurodegenerative conditions can induce the region-specific emergence of cell states relevant to their pathogenesis. To identify such phenomena, we generated a spatiotemporal transcriptomic atlas of mice infected with the RML prion strain. Thalamus and cerebellum experienced severe neuronal loss, developed intense microgliosis and, starting from 30 weeks post-inoculation, accumulated a novel microglial subpopulation characterized by strong expression of Glycoprotein non-metastatic melanoma protein B (Gpnmb). Elevated GPNMB levels were detected in the cerebrospinal fluid of sCJD patients, suggesting its possible usefulness as a biomarker of disease progression. The transcriptional profile of Gpnmb+ microglia reflected a state of enhanced phagocytic activity with upregulation of genes associated with lysosomal function, including vacuolar ATPase V0 domain subunit d2 (Atp6v0d2) and Galectin-3 (Lgals3). In microglia-like murine BV2 cells, Gpnmb upregulation was induced by soluble find-me signals released during apoptosis, but not by apoptotic bodies or prion accumulation. Likewise, human iPSC-derived microglia showed marked upregulation of GPNMB when co-cultured with apoptotic human neurons. Gpnmb ablation impaired the ability of BV2 cells to clear apoptotic cells, underscoring its role in maintaining microglial phagocytosis. Our findings define Gpnmb microglia as a distinct, apoptosis-driven phagocytic state, linking neuronal loss to microglial activation and positioning it as a key regulator of microglial responses to prion propagation.

neuroscience↗

bioRxiv2024

TFAP2C and HNRNPK control mTOR cell metabolism and prion propagation

Heterogeneous Nuclear Ribonucleoprotein K (hnRNP K) is a limiting factor for prion propagation. However, little is known about the function of hnRNP K except that it is essential to cell survival. Here, we performed a synthetic-viability CRISPR ablation screen to identify epistatic interactors of HNRNPK. We found that deletion of Transcription Factor AP-2{gamma} (TFAP2C) suppressed the death of hnRNP K-depleted LN-229 and U-251 MG cells, whereas its overexpression hypersensitized cells to hnRNP K loss. HNRNPK ablation decreased cellular ATP, downregulated genes related to lipid and glucose metabolism, and enhanced autophagy. Co-occurrent deletion of TFAP2C reversed these effects, restoring transcriptional balance and alleviating energy deficiency. We linked HNRNPK and TFAP2C interaction to mTOR signaling, observing that HNRNPK ablation inhibited mTORC1 activity through downregulation of mTOR and Rptor, while TFAP2C overexpression enhanced mTORC1 downstream functions. In prion-infected cells, TFAP2C activation reduced prion levels and countered the increased prion propagation caused by HNRNPK suppression. Short-term pharmacological inhibition of mTOR also elevated prion levels and partially mimicked the effects of HNRNPK silencing. Our study identifies TFAP2C as a genetic interactor of HNRNPK, implicates their roles in mTOR metabolic regulation, and establishes a causative link between these activities and prion propagation.

cell biology↗