المؤلفون: Camille Peron, Andrea Cavaliere, Chiara Fasano, Angelo Iannielli, Manuela Spagnolo, Andrea Legati, Maria Nicol Colombo, Ambra Rizzo, Francesca L. Sciacca, Valerio Carelli, Vania Broccoli, Costanza Lamperti, Valeria Tiranti

المصدر: Stem Cell Research, Vol 77, Iss , Pp 103406- (2024)

مصطلحات موضوعية: Biology (General), QH301-705.5

الوصف: Leber hereditary optic neuropathy (LHON) is one of the most common mitochondrial illness, causing retinal ganglion cell degeneration and central vision loss. It stems from point mutations in mitochondrial DNA (mtDNA), with key mutations being m.3460G > A, m.11778G > A, and m.14484 T > C. Fibroblasts from identical twins, sharing m.14484 T > C and m.10680G > A variants each with 70 % heteroplasmy, were used to generate iPSC lines. Remarkably, one twin, a LHON patient, displayed symptoms, while the other, a carrier, remained asymptomatic. These iPSCs offer a valuable tool for studying factors influencing disease penetrance and unravelling the role of m.10680G > A, which is still debated.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S1873506124001041Test; https://doaj.org/toc/1873-5061Test

الوصول الحر: https://doaj.org/article/ef6d717e79284050a843ebbd78811d1eTest

المؤلفون: Muyesier Maimaitili, Muwan Chen, Fabia Febbraro, Ekin Ucuncu, Rachel Kelly, Jonathan Christos Niclis, Josefine Rågård Christiansen, Noëmie Mermet-Joret, Dragos Niculescu, Johanne Lauritsen, Angelo Iannielli, Ida H. Klæstrup, Uffe Birk Jensen, Per Qvist, Sadegh Nabavi, Vania Broccoli, Anders Nykjær, Marina Romero-Ramos, Mark Denham

المصدر: Nature Communications, Vol 14, Iss 1, Pp 1-23 (2023)

مصطلحات موضوعية: Science

الوصف: Abstract Current differentiation protocols for generating mesencephalic dopaminergic (mesDA) neurons from human pluripotent stem cells result in grafts containing only a small proportion of mesDA neurons when transplanted in vivo. In this study, we develop lineage-restricted undifferentiated stem cells (LR-USCs) from pluripotent stem cells, which enhances their potential for differentiating into caudal midbrain floor plate progenitors and mesDA neurons. Using a ventral midbrain protocol, 69% of LR-USCs become bona fide caudal midbrain floor plate progenitors, compared to only 25% of human embryonic stem cells (hESCs). Importantly, LR-USCs generate significantly more mesDA neurons under midbrain and hindbrain conditions in vitro and in vivo. We demonstrate that midbrain-patterned LR-USC progenitors transplanted into 6-hydroxydopamine-lesioned rats restore function in a clinically relevant non-pharmacological behavioral test, whereas midbrain-patterned hESC-derived progenitors do not. This strategy demonstrates how lineage restriction can prevent the development of undesirable lineages and enhance the conditions necessary for mesDA neuron generation.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2041-1723Test

الوصول الحر: https://doaj.org/article/cabafdbe91e44ff087afd66e98f70397Test

المؤلفون: Giorgia Maria Ferlazzo, Anna Maria Gambetta, Sonia Amato, Noemi Cannizzaro, Silvia Angiolillo, Mattia Arboit, Linda Diamante, Elena Carbognin, Patrizia Romani, Federico La Torre, Elena Galimberti, Florian Pflug, Mirko Luoni, Serena Giannelli, Giuseppe Pepe, Luca Capocci, Alba Di Pardo, Paola Vanzani, Lucio Zennaro, Vania Broccoli, Martin Leeb, Enrico Moro, Vittorio Maglione, Graziano Martello

المصدر: Nature Communications, Vol 14, Iss 1, Pp 1-24 (2023)

مصطلحات موضوعية: Science

الوصف: Abstract Huntington’s disease (HD) is a neurodegenerative disorder caused by CAG-repeat expansions in the huntingtin (HTT) gene. The resulting mutant HTT (mHTT) protein induces toxicity and cell death via multiple mechanisms and no effective therapy is available. Here, we employ a genome-wide screening in pluripotent mouse embryonic stem cells (ESCs) to identify suppressors of mHTT toxicity. Among the identified suppressors, linked to HD-associated processes, we focus on Metal response element binding transcription factor 1 (Mtf1). Forced expression of Mtf1 counteracts cell death and oxidative stress caused by mHTT in mouse ESCs and in human neuronal precursor cells. In zebrafish, Mtf1 reduces malformations and apoptosis induced by mHTT. In R6/2 mice, Mtf1 ablates motor defects and reduces mHTT aggregates and oxidative stress. Our screening strategy enables a quick in vitro identification of promising suppressor genes and their validation in vivo, and it can be applied to other monogenic diseases.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2041-1723Test

الوصول الحر: https://doaj.org/article/c1ac1fcec269480f96fa0a3e4fd588a3Test

المؤلفون: Mattia Zaghi, Fabiana Longo, Luca Massimino, Alicia Rubio, Simone Bido, Pietro Giuseppe Mazzara, Edoardo Bellini, Federica Banfi, Paola Podini, Francesca Maltecca, Alessio Zippo, Vania Broccoli, Alessandro Sessa

المصدر: Molecular Autism, Vol 14, Iss 1, Pp 1-16 (2023)

مصطلحات موضوعية: Neurology. Diseases of the nervous system, RC346-429

الوصف: Abstract Background Neurodevelopmental disorders (NDDs) are heterogeneous conditions due to alterations of a variety of molecular mechanisms and cell dysfunctions. SETD5 haploinsufficiency leads to NDDs due to chromatin defects. Epigenetic basis of NDDs has been reported in an increasing number of cases while mitochondrial dysfunctions are more common within NDD patients than in the general population. Methods We investigated in vitro neural stem cells as well as the brain of the Setd5 haploinsufficiency mouse model interrogating its transcriptome, analyzing mitochondrial structure, biochemical composition, and dynamics, as well as mitochondrial functionality. Results Mitochondrial impairment is facilitated by transcriptional aberrations originated by the decrease of the SETD5 enzyme. Low levels of SETD5 resulted in fragmented mitochondria, reduced mitochondrial membrane potential, and ATP production both in neural precursors and neurons. Mitochondria were also mislocalized in mutant neurons, with reduced organelles within neurites and synapses. Limitations We found several defects in the mitochondrial compartment; however, we can only speculate about their position in the hierarchy of the pathological mechanisms at the basis of the disease. Conclusions Our study explores the interplay between chromatin regulation and mitochondria functions as a possible important aspect of SETD5-associated NDD pathophysiology. Our data, if confirmed in patient context, suggest that the mitochondrial activity and dynamics may represent new therapeutic targets for disorders associated with the loss of SETD5.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2040-2392Test

الوصول الحر: https://doaj.org/article/41a9f2cb20ba4eb3b3a5cb5d759d70b4Test

المؤلفون: Mattia Zaghi, Federica Banfi, Luca Massimino, Monica Volpin, Edoardo Bellini, Simone Brusco, Ivan Merelli, Cristiana Barone, Michela Bruni, Linda Bossini, Luigi Antonio Lamparelli, Laura Pintado, Deborah D’Aliberti, Silvia Spinelli, Luca Mologni, Gaia Colasante, Federica Ungaro, Jean-Michel Cioni, Emanuele Azzoni, Rocco Piazza, Eugenio Montini, Vania Broccoli, Alessandro Sessa

المصدر: Nature Communications, Vol 14, Iss 1, Pp 1-21 (2023)

مصطلحات موضوعية: Science

الوصف: Abstract Within the chromatin, distal elements interact with promoters to regulate specific transcriptional programs. Histone acetylation, interfering with the net charges of the nucleosomes, is a key player in this regulation. Here, we report that the oncoprotein SET is a critical determinant for the levels of histone acetylation within enhancers. We disclose that a condition in which SET is accumulated, the severe Schinzel-Giedion Syndrome (SGS), is characterized by a failure in the usage of the distal regulatory regions typically employed during fate commitment. This is accompanied by the usage of alternative enhancers leading to a massive rewiring of the distal control of the gene transcription. This represents a (mal)adaptive mechanism that, on one side, allows to achieve a certain degree of differentiation, while on the other affects the fine and corrected maturation of the cells. Thus, we propose the differential in cis-regulation as a contributing factor to the pathological basis of SGS and possibly other the SET-related disorders in humans.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2041-1723Test

الوصول الحر: https://doaj.org/article/dfd430e45aab49efbf05d9df651f99c5Test

المؤلفون: Alessia Muscò, Davide Martini, Matteo Digregorio, Vania Broccoli, Massimiliano Andreazzoli

المصدر: Genes, Vol 15, Iss 4, p 492 (2024)

مصطلحات موضوعية: dark genes, neurodevelopmental disorders, eye development, ASD, ADHD, Genetics, QH426-470

الوصف: Haploinsufficiency of the PRR12 gene is implicated in a human neuro-ocular syndrome. Although identified as a nuclear protein highly expressed in the embryonic mouse brain, PRR12 molecular function remains elusive. This study explores the spatio-temporal expression of zebrafish PRR12 co-orthologs, prr12a and prr12b, as a first step to elucidate their function. In silico analysis reveals high evolutionary conservation in the DNA-interacting domains for both orthologs, with significant syntenic conservation observed for the prr12b locus. In situ hybridization and RT-qPCR analyses on zebrafish embryos and larvae reveal distinct expression patterns: prr12a is expressed early in zygotic development, mainly in the central nervous system, while prr12b expression initiates during gastrulation, localizing later to dopaminergic telencephalic and diencephalic cell clusters. Both transcripts are enriched in the ganglion cell and inner neural layers of the 72 hpf retina, with prr12b widely distributed in the ciliary marginal zone. In the adult brain, prr12a and prr12b are found in the cerebellum, amygdala and ventral telencephalon, which represent the main areas affected in autistic patients. Overall, this study suggests PRR12’s potential involvement in eye and brain development, laying the groundwork for further investigations into PRR12-related neurobehavioral disorders.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2073-4425/15/4/492Test; https://doaj.org/toc/2073-4425Test

الوصول الحر: https://doaj.org/article/273b19cb3648403aae33b15c5c17fd08Test

المؤلفون: Ivana Barravecchia, Chiara De Cesari, Viviana Guadagni, Giovanni Signore, Edoardo Bertolini, Serena Gea Giannelli, Francesca Scebba, Davide Martini, Mario Enrico Pè, Vania Broccoli, Massimiliano Andreazzoli, Debora Angeloni, Gian Carlo Demontis

المصدر: Scientific Reports, Vol 13, Iss 1, Pp 1-17 (2023)

مصطلحات موضوعية: Medicine, Science

الوصف: Abstract In proliferating multipotent retinal progenitors, transcription factors dynamics set the fate of postmitotic daughter cells, but postmitotic cell fate plasticity driven by extrinsic factors remains controversial. Transcriptome analysis reveals the concurrent expression by postmitotic rod precursors of genes critical for the Müller glia cell fate, which are rarely generated from terminally-dividing progenitors as a pair with rod precursors. By combining gene expression and functional characterisation in single cultured rod precursors, we identified a time-restricted window where increasing cell culture density switches off the expression of genes critical for Müller glial cells. Intriguingly, rod precursors in low cell culture density maintain the expression of genes of rod and glial cell fate and develop a mixed rod/Muller glial cells electrophysiological fingerprint, revealing rods derailment toward a hybrid rod-glial phenotype. The notion of cell culture density as an extrinsic factor critical for preventing rod-fated cells diversion toward a hybrid cell state may explain the occurrence of hybrid rod/MG cells in the adult retina and provide a strategy to improve engraftment yield in regenerative approaches to retinal degenerative disease by stabilising the fate of grafted rod precursors.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2045-2322Test

الوصول الحر: https://doaj.org/article/dab0c647c8294922ac3b468facc4ad7eTest

المؤلفون: Angelo Iannielli, Mirko Luoni, Serena Gea Giannelli, Rosangela Ferese, Gabriele Ordazzo, Matteo Fossati, Andrea Raimondi, Felipe Opazo, Olga Corti, Jochen H. M. Prehn, Stefano Gambardella, Ronald Melki, Vania Broccoli

المصدر: Cell Death and Disease, Vol 13, Iss 10, Pp 1-16 (2022)

مصطلحات موضوعية: Cytology, QH573-671

الوصف: Abstract Triplication of the SNCA gene, encoding the protein alpha-Synuclein (αSyn), is a rare cause of aggressive and early-onset parkinsonism. Herein, we generated iPSCs from two siblings with a recently described compact SNCA gene triplication and suffering from severe motor impairments, psychiatric symptoms, and cognitive deterioration. Using CRISPR/Cas9 gene editing, each SNCA copy was inactivated by targeted indel mutations generating a panel of isogenic iPSCs with a decremental number from 4 down to none of functional SNCA gene alleles. We differentiated these iPSC lines in midbrain dopaminergic (DA) neuronal cultures to characterize αSyn aggregation in native and seeded conditions and evaluate its associated cellular dysfunctions. Utilizing a new nanobody-based biosensor combined with super-resolved imaging, we were able to visualize and measure αSyn aggregates in early DA neurons in unstimulated conditions. Calcium dysregulation and mitochondrial alterations were the first pathological signs detectable in early differentiated DA neuronal cultures. Accelerated αSyn aggregation was induced by exposing neurons to structurally well-characterized synthetic αSyn fibrils. 4xSNCA DA neurons showed the highest vulnerability, which was associated with high levels of oxidized DA and amplified by TAX1BP1 gene disruption. Seeded DA neurons developed large αSyn deposits whose morphology and internal constituents resembled Lewy bodies commonly observed in Parkinson’s disease (PD) patient brain tissues. These findings provide strong evidence that this isogenic panel of iPSCs with SNCA multiplications offers a remarkable cellular platform to investigate mechanisms of PD and validate candidate inhibitors of native and seeded αSyn aggregation.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2041-4889Test

الوصول الحر: https://doaj.org/article/00822ab787424b9bba7317ec699b92daTest

المؤلفون: Greta Rossi, Gabriele Ordazzo, Niccolò N Vanni, Valerio Castoldi, Angelo Iannielli, Dario Di Silvestre, Edoardo Bellini, Letizia Bernardo, Serena G Giannelli, Mirko Luoni, Sharon Muggeo, Letizia Leocani, PierLuigi Mauri, Vania Broccoli

المصدر: eLife, Vol 12 (2023)

مصطلحات موضوعية: eye, retinal development, glial cell, astrocytes, Medicine, Science, Biology (General), QH301-705.5

الوصف: Wolfram syndrome 1 (WS1) is a rare genetic disorder caused by mutations in the WFS1 gene leading to a wide spectrum of clinical dysfunctions, among which blindness, diabetes, and neurological deficits are the most prominent. WFS1 encodes for the endoplasmic reticulum (ER) resident transmembrane protein wolframin with multiple functions in ER processes. However, the WFS1-dependent etiopathology in retinal cells is unknown. Herein, we showed that Wfs1 mutant mice developed early retinal electrophysiological impairments followed by marked visual loss. Interestingly, axons and myelin disruption in the optic nerve preceded the degeneration of the retinal ganglion cell bodies in the retina. Transcriptomics at pre-degenerative stage revealed the STAT3-dependent activation of proinflammatory glial markers with reduction of the homeostatic and pro-survival factors glutamine synthetase and BDNF. Furthermore, label-free comparative proteomics identified a significant reduction of the monocarboxylate transport isoform 1 (MCT1) and its partner basigin that are highly enriched on retinal glia and myelin-forming oligodendrocytes in optic nerve together with wolframin. Loss of MCT1 caused a failure in lactate transfer from glial to neuronal cell bodies and axons leading to a chronic hypometabolic state. Thus, this bioenergetic impairment is occurring concurrently both within the axonal regions and cell bodies of the retinal ganglion cells, selectively endangering their survival while impacting less on other retinal cells. This metabolic dysfunction occurs months before the frank RGC degeneration suggesting an extended time-window for intervening with new therapeutic strategies focused on boosting retinal and optic nerve bioenergetics in WS1.

وصف الملف: electronic resource

العلاقة: https://elifesciences.org/articles/81779Test; https://doaj.org/toc/2050-084XTest

الوصول الحر: https://doaj.org/article/587df71ad8444471a703dfcbc9ffb880Test

المؤلفون: Marta Biondi, Sarah Tettamanti, Stefania Galimberti, Beatrice Cerina, Chiara Tomasoni, Rocco Piazza, Samantha Donsante, Simone Bido, Vincenzo Maria Perriello, Vania Broccoli, Andrea Doni, Francesco Dazzi, Alberto Mantovani, Gianpietro Dotti, Andrea Biondi, Alice Pievani, Marta Serafini

المساهمون: Biondi, M, Tettamanti, S, Galimberti, S, Cerina, B, Tomasoni, C, Piazza, R, Donsante, S, Bido, S, Maria Perriello, V, Broccoli, V, Doni, A, Dazzi, F, Mantovani, A, Dotti, G, Biondi, A, Pievani, A, Serafini, M

مصطلحات موضوعية: CAR-CIK, Acute Myeloid Leukemia

الوصف: Acute myeloid leukemia (AML) is a hematological malignancy derived from neoplastic myeloid progenitor cells characterized by abnormal clonal proliferation and differentiation. Although novel therapeutic strategies have recently been introduced, the prognosis of AML remains unsatisfactory. So far, the efficacy of chimeric antigen receptor (CAR)-T therapy in AML has been hampered by several factors including the poor accumulation of the blood-injected cells in the leukemia bone marrow (BM) niche, where chemotherapy-resistant leukemic stem cells reside. Thus, we hypothesized that overexpression of CXCR4, whose ligand CXCL12 is highly expressed by BM stromal cells within the niche, could improve T cell homing to the BM and consequently enhance their intimate contact with BM-resident AML cells facilitating disease eradication. Specifically, we engineered conventional CD33.CAR-cytokine induced killer cells (CIKs) with the wild-type CXCR4 and the variant CXCR4R334X, responsible for leukocyte sequestration in the BM of WHIM syndrome patients. Overexpression of both CXCR4wt and CXCR4mut in CD33.CAR-CIKs resulted in significant improvement of chemotaxis toward recombinant CXCL12 or BM stromal cell conditioned medium with no observed impairment of cytotoxic potential in vitro. Moreover, CXCR4-overexpressing CD33.CAR-CIKs showed enhanced in vivo BM homing, associated with a prolonged retention for the CXCR4R334X variant. However, only CD33.CAR-CIKs co-expressing CXCR4wt but not CXCR4mut exerted a more sustained in vivo antileukemic activity and extended animal survival, suggesting a non-canonical role for CXCR4 in modulating CAR-CIK functions independent of BM homing. Taken together, these data suggest that arming CAR-CIKs with CXCR4 may represent a promising strategy for increasing their therapeutic potential for AML.

وصف الملف: STAMPA

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/36787509; info:eu-repo/semantics/altIdentifier/wos/WOS:001053602500001; volume:141; issue:21; firstpage:2587; lastpage:2598; numberofpages:12; journal:BLOOD; https://hdl.handle.net/10281/414788Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85152748326

الإتاحة: https://doi.org/10.1182/blood.2022018330Test
https://hdl.handle.net/10281/414788Test