Biology subjects

Morabito, G.

Publications and source records attributed to Morabito, G..

4 recordsLinked to original sources

bioRxiv2023

Assessing Age-Related Decline in Anti-Bacterial Immune Responses Using an Ex-vivo Assay in African Turquoise Killifish (Nothobranchius furzeri)

Ex-vivo immune functional assays serve as essential tools for uncovering the intricate molecular mechanisms behind immune responses, both intrinsic and extrinsic, across a diverse spectrum of conditions and interventions. In this study, we devised an innovative assay aimed at quantifying anti-bacterial responses in immune cells from the primary immune organ of small teleosts. Our model of choice is the the African turquoise killifish (Nothobranchius furzeri), which has emerged as a prominent model system in the field of biology of aging, because of its natural short lifespan and the broad range of aging-related dysfunctions. Using our novel ex vivo assay, we tested age-associated differences in immune cell functionality, measured as anti-bacterial responses in young and aged turquoise killifish. Our results show that the ability to inhibit bacterial growth declines in cells extracted from aged killifish, compared to young killifish. This ex-vivo assay, designed for rapid measures of immune effector responses, holds the potential to be a scalable tool for assessing the cellular and molecular basis for anti-microbial immune responses under a range of interventions.

immunology↗

bioRxiv2023

Spontaneous onset of cellular markers of inflammation and genome instability during aging in the immune niche of the naturally short-lived turquoise killifish (Nothobranchius furzeri)

Turquoise killifish (Nothobranchius furzeri) are naturally short-lived vertebrates, recapitulating several aspects of human aging, including protein aggregation, telomere shortening, cellular senescence, and declined antibody diversity. The mechanistic causes of systemic aging in killifish are still poorly understood. Here we ask whether killifish undergo significant age-dependent changes in the main hematopoietic organ, which together could contribute to systemic aging. To characterize immune aging in killifish, we employed single-cell RNA sequencing, proteomics, cytometry and a functional in vitro assay. Our data indicate how old killifish display increased inflammatory markers, and while immune cells from adult killifish display increased markers of proliferation and replication-independent DNA repair in progenitor-like cell clusters, progenitors from old killifish display extensive markers of DNA double-strand breaks. In less than 10 weeks, killifish undergo several dramatic spontaneous aging-related changes in the immune niche, which could be functionally linked with its extensive systemic aging and serve as targets for anti-aging interventions.

immunology↗

bioRxiv2019

Whole brain delivery of an instability-prone Mecp2 transgene improves behavioral and molecular pathological defects in mouse models of Rett syndrome

Rett syndrome (RTT) is an incurable neurodevelopmental disorder caused by mutations in the gene encoding for methyl-CpG binding-protein 2 (MeCP2). Gene therapy for this disease presents inherent hurdles since MECP2 is expressed throughout the brain and its duplication leads to severe neurological conditions as well. However, the recent introduction of AAV-PHP.eB, an engineered capsid with an unprecedented efficiency in crossing the blood-brain barrier upon intravenous injection, has provided an invaluable vehicle for gene transfer in the mouse nervous system. Herein, we use AAV-PHP.eB to deliver an instability-prone Mecp2 (iMecp2) transgene cassette which, increasing RNA destabilization and inefficient protein translation of the viral Mecp2 transgene, limits supraphysiological Mecp2 protein levels in transduced neural tissues. Intravenous injections of the PHP.eB-iMecp2 virus in symptomatic male and female Mecp2 mutant mice significantly ameliorated the disease progression with improved locomotor activity, coordination, lifespan and normalization of altered gene expression and mTOR signaling. Remarkably, PHP.eB-iMecp2 administration did not result in severe toxicity effects either in female Mecp2 mutant or in wild-type animals. In contrast, we observed a strong immune response to the transgene in treated male Mecp2 mutant mice that was overcome by immunosuppression. Overall, PHP.eB-mediated delivery of the iMecp2 cassette provided widespread and efficient gene transfer maintaining physiological Mecp2 protein levels in the brain. This combination defines a novel viral system with significant therapeutic efficacy and increased safety which can contribute to overcome the hurdles that are delaying clinical applications of gene therapy for RTT.\n\nOne Sentence SummaryGlobal brain transduction of the instability-prone Mecp2 transgene by systemic AAV-PHP.eB administration is both safe and effective in protecting male and female Mecp2 mutant mice from the RTT disease phenotype.

neuroscience↗

bioRxiv2019

Easy Kinetics: a software that simplify enzymes kinetics characterization

Here will be presented the software Easy Kinetics, a publicly available graphical interface that allows rapid evaluation of the main kinetics parameters in an enzyme catalyzed reaction. In contrast to other similar commercial software using algorithms based on non-linear regression models to reach these results, Easy Kinetics is based on a completely different original algorithm, requiring in input the spectrophotometric measurements of {Delta}Abs/min taken twice at only two different substrate concentrations. The results generated show however a significant concordance with those ones obtained with the most common commercial software used for enzyme kinetics characterization, GraphPad Prism 8(C), suggesting that Easy Kinetics can be used for routine tests in enzyme kinetics as an alternative valid software.

biochemistry↗