Biology subjects

Cenini, G.

Publications and source records attributed to Cenini, G..

2 recordsLinked to original sources

bioRxiv2026

Glia-enriched human brain organoids for studying developmental alterations in leukodystrophies

Leukodystrophies (LKD) comprise mostly inherited white matter disorders whose developmental origins and underlying pathomechanisms remain poorly understood. Progress has been limited by restricted access to human primary tissue and by the inability of animal models to fully recapitulate human neurodevelopment. Human induced pluripotent stem cell-derived neural organoids provide an alternative approach, but most culture systems do not capture later, glia-rich developmental stages relevant to these diseases. Recent studies have shown that appropriate patterning cues and long in vitro differentiation can generate neuroglial spheroids comprising multiple neuronal and macroglial populations, including mature oligodendrocytes. Here, we adapt the human oligodendrocyte (hOLS) spheroid platform to establish 3D models of globoid cell leukodystrophy (GLD), 4H leukodystrophy (4H), and Canavan disease (CD). Here we adapt this organoid platform to establish 3D models of globoid cell leukodystrophy (GLD), 4H leukodystrophy (4H), and Canavan disease (CD). Across 150 days of differentiation, hOLS from different genetic backgrounds displayed largely comparable growth kinetics and developmental trajectories, comprising multiple neuroglial cell types with predominantly ventral forebrain and midbrain identities. Comparison of LKD and control hOLS revealed both shared and disease-specific alterations, with prominent transcriptional changes in lysosomal, metabolic, and stress-response pathways. Lineage-resolved pathway analysis further identified signatures across individual LKDs, including mitochondrial and translational stress in GLD glial lineages, disruption of RNA metabolic and cell-cycle pathways in 4H, and prominent astrocyte-centered metabolic dysregulation in CD. Our results establish hOLS as a human 3D in vitro platform for studying the impact of LKD-associated genetic variants on early neural development and for dissecting lineage-specific disease vulnerability.

neuroscience↗

bioRxiv2022

Hsp78-interaction proteome highlights chaperone role during protein aggregation in mitochondria under heat stress

Proteins of the Hsp100 chaperone family support protein homeostasis, the maintenance of protein activity under stress, by refolding aggregated proteins or targeting them for degradation. Hsp78, the ClpB-type mitochondrial member of the Hsp100 family, can be found in lower eukaryotes like yeast. Although Hsp78 has been shown to contribute to protection against elevated temperatures in yeast, the biochemical mechanisms underlying this mitochondria-specific thermotolerance are still largely unclear. To identify endogenous chaperone substrate proteins, we generated an Hsp78-ATPase mutant with a stabilized substrate binding behaviour. We used two SILAC-based quantitative mass spectrometry approaches to analyze the role of Hsp78 during heat stress-induced mitochondrial protein aggregation and disaggregation processes on a proteomic level. In the first setup, Hsp78-interacting polypeptides were identified to reveal the endogenous substrate spectrum of the chaperone. Our analysis revealed that Hsp78 is interacting with a wide variety of proteins related to metabolic functions including energy production and protein synthesis, as well as other chaperones, thus maintaining crucial functions for mitochondrial stress resistance. We compared these interaction data with a second experimental setup that focussed on the on overall aggregation and disaggregation processes in mitochondria under heat stress on a proteomic level. This revealed specific aggregation-prone protein populations and demonstrated the direct quantitative impact of Hsp78 on stress-dependent protein solubility under different conditions. We conclude that Hsp78 together with its cofactors represents a recovery system that protects major mitochondrial metabolic functions during heat stress as well restores protein biogenesis capacity after return to normal conditions.

cell biology↗