Biology subjects

Bano, D.

Publications and source records attributed to Bano, D..

9 recordsLinked to original sources

bioRxiv2026

Reverse engineering Lewy bodies suggests alpha-synuclein pathology as condensatopathy

Understanding how soluble monomeric alpha-synuclein transitions into pathological alpha-synuclein (Lewy bodies) has remained a central challenge in neurodegeneration. Existing models, largely based on treating cells and animals with pre-formed alpha-synuclein fibrils cannot account for the emergence of the first inclusion without pre-existing seeds. Here, we combine in vitro reconstitutions, computational modeling and a minimal cellular system to show that phase separation of alpha-synuclein with an interacting scaffold can recapitulate Lewy body-like structures. An unbiased proteomics screen, live-cell imaging, correlative light-electron microscopy, and soft X-ray tomography show that Lewy body-like structures are associated with redistribution of actin and membrane-bound organelles, with mitochondria accumulating at their periphery in an actin-dependent manner. Structural analyses further indicate that alpha-synuclein condensates provide local environments in which the insoluble beta-sheets can form. Together, these findings support a seed-independent trajectory from soluble alpha-synuclein through condensation and progressive immobilization to Lewy body-like assemblies and, subsequently, insoluble beta-sheet fibrils. This framework offers a new mechanistic route for understanding the disease progression and promises to guide the design of innovative therapeutic interventions.

neuroscience↗

bioRxiv2026

Long-term rapamycin treatment suppresses IL-17-producing gamma delta T cells and blunts neuroinflammation in aging

Aging is the gradual accumulation of structural and functional changes in an organism over time, including immune remodeling and a progressive increase in basal inflammation, or inflammaging. The mTOR pathway is a central driver of aging-related diseases, such as cancer, chronic inflammation and neurodegeneration; pharmacological inhibition with rapamycin is associated with reduced aged-related morbidity and increased lifespan across species. Nonetheless, concerns remain about the use of rapamycin, a well-established immunosuppressant in transplant medicine, as an anti-aging intervention. Here, we evaluated the impact of prolonged low-dose dietary rapamycin on the aging immune system. Treatment did not significantly alter innate or adaptive immune cell populations, including brain resident microglia; however, it attenuated the age-associated accumulation of IL-17-producing {gamma}{delta} T cells, particularly in the peritoneal cavity. After a peripheral inflammatory LPS challenge, circulating IL-17 levels were significantly reduced and correlated with an attenuation of microglia inflammatory phenotype. These findings suggest that prolonged low-dose rapamycin exposure exerts minor systemic immune changes, while selectively limiting age-related {gamma}{delta} T cell expansion and neuroinflammation associated with systemic inflammation.

immunology↗

bioRxiv2025

Integrating simulated and experimental data to identify mitochondrial bioenergetic defects in Parkinson's Disease models

Mitochondrial bioenergetics are vital for ATP production and are associated with several diseases, including Parkinsons Disease. Here, we simulated a computational model of mitochondrial ATP production to interrogate mitochondrial bioenergetics under physiological and pathophysiological conditions, and provide a data resource that can be used to interpret mitochondrial bioenergetics experiments. We first characterised the impact of several common respiratory chain impairments on experimentally-observable bioenergetic parameters. We then established an analysis pipeline to integrate simulations with experimental data and predict the molecular defects underlying experimental bioenergetic phenotypes. We applied the pipeline to data from Parkinsons Disease models. We verified that the impaired bioenergetic profile previously measured in Parkin knockout neurons can be explained by increased mitochondrial uncoupling. We then generated primary cortical neurons from a Pink1 KO mouse model of Parkinsons, and measured reduced OCR capacity and increased resistance to Complex III inhibition. Here, our pipeline predicted that multiple respiratory chain impairments are required to explain this bioenergetic phenotype. Finally, we provide all simulated data as a user-friendly resource that can be used to interpret mitochondrial bioenergetics experiments, predict underlying molecular defects, and inform experimental design. HighlightsO_LIThe complexity of mitochondrial bioenergetics can make experimental data difficult to interpret. C_LIO_LIWe simulated a computational model of mitochondrial bioenergetics in healthy and pathological conditions, and established an analysis pipeline to integrate model simulations with experimental data. C_LIO_LIWe applied the pipeline to data from Parkinsons Disease models to predict the molecular defects underlying Parkinsons-related pathology. C_LIO_LIWe provide all outputs in a user-friendly Excel file, which serves as a valuable resource to the community for insight into the effects of pathology on mitochondrial bioenergetics and for interpretation of experimental results. C_LI

neuroscience↗

bioRxiv2025

Manipulation of the Extracellular Vesicle Biogenesis Pathway in Paternal Somatic Cells Contributes to the Phenotypic Fate of the Next Generation

Human and animal studies have shown that risk factors for certain diseases are linked to our fathers physiological and psychological condition. So far, mainly in vitro evidence indicates extracellular vesicles (EVs) as one of the potential mediators of these paternal effects. In a full in vivo model system, we demonstrate that targeting the EV biogenesis pathway within somatic cells of the male Drosophila melanogaster reproductive tract induces sex-specific phenotypic changes in the progeny. These findings highlight the critical role of the EV pathway in paternal somatic cells in shaping the phenotypic destiny of the next generation and underscore the necessity for further investigations into the mechanisms of somatic to germline information transfer.

physiology↗

bioRxiv2024

Dynamics of microglia-glioblastoma crosstalk at the far infiltration zone

The interaction of glioblastoma (GB) and microglia is critical due to its implications for tumor progression, immune response modulation, and potential therapeutic strategies. However, the role of microglia in GB pathogenesis remains unclear, especially regarding the in vivo dynamics of their interplay. Performing three-photon imaging in an autochthonous, immunocompetent mouse GB model, we examined tumor/microglia dynamics within previously inaccessible regions at the GB far infiltration zone in the corpus callosum. Initially, microglia increased tissue surveillance upon encountering GB-cells in sparsely infiltrated areas. In contrast, when GB-cell density increased, microglia reduced surveillance, suggesting a biphasic response to tumor invasion. Additionally, microglia were not uniformly attracted to infiltrating GB-cells; only a subset moved directionally toward them within a defined spatial range. This study provides insight into the heterogeneity of the immune response to tumor invasion and the dynamics of microglia-GB interactions in vivo. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=124 SRC="FIGDIR/small/614016v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@24b1d8org.highwire.dtl.DTLVardef@1180609org.highwire.dtl.DTLVardef@346dbforg.highwire.dtl.DTLVardef@1144dd_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2024

Interaction with AK2A links AIFM1 to cellular energy metabolism

Apoptosis inducing factor 1 (AIFM1) is a flavoprotein essential for mitochondrial function and biogenesis. Its interaction with MIA40, the central component of the mitochondrial disulfide relay, accounts for some, but not all effects of AIFM1 loss. Our high-confidence AIFM1 interactome revealed novel interaction partners of AIFM1. For one of these interactors, adenylate kinase 2 (AK2), an essential enzyme maintaining cellular adenine nucleotide pools, AIFM1 binding specifically stabilized the isoform AK2A via interaction with its C-terminus. High resolution cryo-EM and biochemical analyses showed that both, MIA40 and AK2A bind AIFM1s C-terminal {beta}-strand, enhancing NADH oxidoreductase activity by locking an active, dimer conformation and, in the case of MIA40, affecting the cofactor binding site. The AIFM1-AK2A interaction is crucial during respiratory conditions. We further identified ADP/ATP translocases and the ATP synthase as AIFM1 interactors, emphasizing its important regulatory role as a central, organizing platform in energy metabolism.

biochemistry↗

bioRxiv2023

TMEM65-dependent Ca2+ extrusion safeguards mitochondrial homeostasis

The bidirectional transport of Ca2+ into and out of mitochondria is a conserved biological process controlling multiple events, including metabolism, signaling, and cell fate. In the presence of membrane potential driving mitochondrial Ca2+ accumulation, transient changes of [Ca2+]mt in response to cytosolic [Ca2+] variations are ensured by a molecular machinery for Ca2+ influx and efflux embedded in the inner mitochondrial membrane (IMM). While it is well established that influx relies on the Mitochondrial Calcium Uniporter (MCU), efflux was expected to be molecularly diversified, given the occurrence of functionally different exchange pathways with either Na+ or H+1. Accordingly, dedicated transporters ensure proper Ca2+ homeostasis and tightly regulated mitochondrial bioenergetics, but the process is not yet fully elucidated. We here demonstrate that TMEM65, a protein with an unknown biological function, is a fundamental component of the Ca2+ efflux machinery of mitochondria. As the MCU, TMEM65 has a broad tissue expression and localizes to the IMM. Its overexpression dramatically enhances Na+- and Li+-dependent mitochondrial Ca2+ extrusion, which is abrogated by the pharmacological inhibitor CGP-37157. Conversely, its downregulation chronically elevates resting mitochondrial Ca2+ levels and inhibits efficient Ca2+ efflux upon cellular activation, culminating in mitochondrial calcium overload and causing organelle dysfunction. Since TMEM65 has been associated with a severe human mitochondrial disease2, we deleted the TMEM65 homologues in Caenorhabditis elegans (CeTMEM65). While the two CeTMEM65 orthologs are dispensable for the survival at permissive growing conditions, their loss undermines embryonic developments when eggs are exposed to mild temperature-stress. In this regard, we find that CeTMEM65 (null) alleles cause necrotic lesions that are suppressed by inhibiting the mitochondrial calcium uniporter MCU-1. Overall, these results unambiguously assign a primary role in mitochondrial Ca2+ homeostasis to the orphan protein TMEM65. More importantly, our findings describe a novel molecular component that may be relevant in pathological settings in which excessive mitochondrial Ca2+ accumulation critically contribute to degenerative pathways.

biochemistry↗

bioRxiv2022

Deep Phenotyping and Lifetime Trajectories Reveal Limited Effects of Longevity Regulators on the Aging Process in C57BL/6J Mice

Current concepts regarding the biology of aging are based on studies aimed at identifying factors regulating natural lifespan. However, lifespan as a sole proxy measure for aging can be of limited value because it may be restricted by specific sets of pathologies, rather than by general physiological decline. Here, we employed large-scale phenotyping to analyze hundreds of phenotypes and thousands of molecular markers across tissues and organ systems in a single study of aging male C57BL/6J mice. For each phenotype, we established lifetime profiles to determine when age-dependent phenotypic change is first detectable relative to the young adult baseline. We examined central genetic and environmental lifespan regulators (putative anti-aging interventions, PAAIs; the following PAAIs were examined: mTOR loss-of-function, loss-of-function in growth hormone signaling, dietary restriction) for a possible countering of the signs and symptoms of aging. Importantly, in our study design, we included young treated groups of animals, subjected to PAAIs prior to the onset of detectable age-dependent phenotypic change. In parallel to our studies in mice, we assessed genetic variants for their effects on age-sensitive phenotypes in humans. We observed that, surprisingly, many PAAI effects influenced phenotypes long before the onset of detectable age-dependent changes, rather than altering the rate at which these phenotypes developed with age. Accordingly, this subset of PAAI effects does not reflect a targeting of age-dependent phenotypic change. Overall, our findings suggest that comprehensive phenotyping, including the controls built in our study, is critical for the investigation of PAAIs as it facilitates the proper interpretation of the mechanistic mode by which PAAIs influence biological aging. HighlightsO_LIPhenotyping at scale defines lifetime trajectories of age-dependent changes in C57BL/6J mice C_LIO_LICentral genetic and environmental lifespan regulators (putative anti-aging interventions; PAAIs) influence age-sensitive phenotypes (ASPs) often long before the appearance of age-dependent changes in these ASPs C_LIO_LICorresponding genetic variants in humans also have age-independent effects C_LIO_LIMany PAAI effects shift the baseline of ASPs rather than slowing their rate of change C_LI

physiology↗

bioRxiv2020

Comparative profiling of N-respirasomes predicts aberrant mitochondrial bioenergetics at single-cell resolution

AO_SCPLOWBSTRACTC_SCPLOWMitochondria sustain the energy demand of the cell. The composition and functional state of the mitochondrial oxidative phosphorylation system are informative indicators of organelle homeostasis and bioenergetic capacity. Here we describe a highly sensitive and reproducible method for single-cell visualization and quantification of mitochondrial respiratory supercomplexes as a novel means of measuring mitochondrial respiratory chain integrity. We apply a proximity ligation assay (PLA) and perform comparative studies of mitochondrial CI, CIII and CIV-containing supercomplexes (or N-respirasomes) in fixed human and mouse brain tissues, tumorigenic cells, iPSCs and iPSC-derived NPCs and neurons. Our optimized approach enables a quantitative in-situ assessment of even subtle mitochondrial lesions associated with aberrant respiration. By combining quantitative proteomics with single cell imaging analysis, we also report the mechanistic contribution of the MICOS complex subunit CHCHD3 in regulating N-respirasomes. Overall, our PLA-based profiling of N-respirasomes establishes a sensitive and complementary technique for detecting cell-type specific mitochondrial perturbations in fixed materials.

cell biology↗