A5091158204- Mitochondrial Function and Pathology
- Genetics and Neurodevelopmental Disorders
- Genetic Neurodegenerative Diseases
- Pluripotent Stem Cells Research
- Metabolism and Genetic Disorders
- Neurogenesis and neuroplasticity mechanisms
- CRISPR and Genetic Engineering
- Autophagy in Disease and Therapy
- RNA regulation and disease
- Virus-based gene therapy research
- Machine Learning in Bioinformatics
- Endoplasmic Reticulum Stress and Disease
- Parkinson's Disease Mechanisms and Treatments
- RNA Interference and Gene Delivery
- Peptidase Inhibition and Analysis
- Retinal Development and Disorders
- Cell death mechanisms and regulation
- Neurological disorders and treatments
- Herpesvirus Infections and Treatments
- RNA Research and Splicing
- Cellular transport and secretion
- Nerve injury and regeneration
- Developmental Biology and Gene Regulation
- Genetics, Aging, and Longevity in Model Organisms
- Neuroinflammation and Neurodegeneration Mechanisms
Vita-Salute San Raffaele University
2019-2024
Neuroscience Institute
2013-2024
National Research Council
2016-2024
Istituti di Ricovero e Cura a Carattere Scientifico
2022-2024
Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele
2023-2024
San Raffaele University of Rome
2016-2022
Accademia Albertina delle Belle Arti
2013
University of Turin
2013
Dysfunctions in mitochondrial dynamics and metabolism are common pathological processes associated with Parkinson's disease (PD). It was recently shown that an inherited form of PD dementia is caused by mutations the OPA1 gene, which encodes for a key player fusion structure. iPSC-derived neural cells from these patients exhibited severe fragmentation, respiration impairment, ATP deficits, heightened oxidative stress. Reconstitution normal levels PD-derived normalized mitochondria morphology...
Mutations in PARK2, encoding the E3 ubiquitin protein ligase Parkin, are a common cause of autosomal recessive Parkinson's disease (PD). Loss PARK2 function compromises mitochondrial quality by affecting biogenesis, bioenergetics, dynamics, transport and turnover. We investigated impact dysfunction on endoplasmic reticulum (ER)-mitochondria interface, which mediates calcium (Ca2+) exchange between two compartments is essential for Parkin-dependent mitophagy. Confocal electron microscopy...
Leber's hereditary optic neuropathy (LHON), a disease associated with mitochondrial DNA mutation, is characterized by blindness due to degeneration of retinal ganglion cells (RGCs) and their axons, which form the nerve.We show that sustained pathological autophagy compartment-specific mitophagy activity affects LHON patient-derived cybrids, as well induced pluripotent-stem-cell-derived neurons.This variably counterbalanced compensatory mitobiogenesis.The aberrant quality control disrupts...
Abstract Friedreich’s ataxia (FRDA) is an autosomal-recessive neurodegenerative and cardiac disorder which occurs when transcription of the FXN gene silenced due to excessive expansion GAA repeats into its first intron. Herein, we generate dorsal root ganglia organoids (DRG organoids) by in vitro differentiation human iPSCs. Bulk single-cell RNA sequencing show that DRG present a transcriptional signature similar native DRGs display main peripheral sensory neuronal glial cell subtypes....
Rett syndrome is an incurable neurodevelopmental disorder caused by mutations in the gene encoding for methyl-CpG binding-protein 2 (MeCP2). Gene therapy this disease presents inherent hurdles since MECP2 expressed throughout brain and its duplication leads to severe neurological conditions as well. Herein, we use AAV-PHP.eB deliver instability-prone Mecp2 (iMecp2) transgene cassette which, increasing RNA destabilization inefficient protein translation of viral transgene, limits...
Stem cell-derived neurons are generally obtained in mass cultures that lack both spatial organization and any meaningful connectivity. We implement a microfluidic system for long-term culture of human with patterned projections synaptic terminals. Co-culture midbrain dopaminergic striatal medium spiny on the microchip establishes an orchestrated nigro-striatal circuitry functional synapses. use this platform to dissect mitochondrial dysfunctions associated genetic form Parkinson's disease...
The CRISPR/Cas9 system is a rapid and customizable tool for gene editing in mammalian cells. In particular, this approach has widely opened new opportunities genetic studies neurological disease. Human neurons can be differentiated vitro from hPSC (human Pluripotent Stem Cells), hNPCs Neural Precursor Cells) or even directly reprogrammed fibroblasts. Here, we described platform which enables, efficient CRISPR/Cas9-mediated genome targeting simultaneously with three different paradigms...
Wolfram syndrome 1 (WS1) is a rare genetic disorder caused by mutations in the WFS1 gene leading to wide spectrum of clinical dysfunctions, among which blindness, diabetes, and neurological deficits are most prominent. encodes for endoplasmic reticulum (ER) resident transmembrane protein wolframin with multiple functions ER processes. However, -dependent etiopathology retinal cells unknown. Herein, we showed that Wfs1 mutant mice developed early electrophysiological impairments followed...
Although adeno-associated virus 9 (AAV9) has been highly exploited as delivery platform for gene-based therapies, its efficacy is hampered by low efficiency in crossing the adult blood-brain barrier (BBB) and pronounced targeting to liver upon intravenous delivery. We generated a new galactose binding-deficient AAV9 peptide display library selected two engineered capsids with enhanced mouse marmoset brains after Interestingly, loss of binding greatly reduced undesired peripheral organs,...
COUP-TFI is an orphan nuclear receptor acting as a strong transcriptional regulator in different aspects of forebrain embryonic development. In this study, we investigated expression and function the mouse olfactory bulb (OB), highly plastic telencephalic region which continuous integration newly generated inhibitory interneurons occurs throughout life. OB belong to populations that originate from distinct progenitor lineages. Here, show expressed tyrosine hydroxylase (TH)-positive...
Abstract Schwann cells (SCs) generate the myelin wrapping of peripheral nerve axons and are promising candidates for cell therapy. However, to date a renewable source SCs is lacking. In this study, we show conversion skin fibroblasts into induced (iSCs) by driving expression two transcription factors, Sox10 Egr2. iSCs resembled primary in global gene profiling PNS identity. vitro , wrapped generating compact sheaths with regular nodal structures. Conversely, from Twitcher mice showed severe...
Abstract Triplication of the SNCA gene, encoding protein alpha-Synuclein (αSyn), is a rare cause aggressive and early-onset parkinsonism. Herein, we generated iPSCs from two siblings with recently described compact gene triplication suffering severe motor impairments, psychiatric symptoms, cognitive deterioration. Using CRISPR/Cas9 editing, each copy was inactivated by targeted indel mutations generating panel isogenic decremental number 4 down to none functional alleles. We differentiated...
The search for disease-modifying agents targeted against Parkinson's disease led us to rationally design a small array of six Anle138b-centered PROTACs, 7a,b, 8a,b and 9a,b, targeting αSynuclein (αSyn) aggregates binding, polyubiquitination by the E3 ligase Cereblon (CRBN), proteasomal degradation. Lenalidomide thalidomide were used as CRBN ligands coupled with amino- azido Anle138b derivatives through flexible linkers coupling reactions (amidation, 'click' chemistry). Four Anle138b-PROTACs,...
Current differentiation protocols for generating mesencephalic dopaminergic (mesDA) neurons from human pluripotent stem cells result in grafts containing only a small proportion of mesDA when transplanted vivo. In this study, we develop lineage-restricted undifferentiated (LR-USCs) cells, which enhances their potential differentiating into caudal midbrain floor plate progenitors and neurons. Using ventral protocol, 69% LR-USCs become bona fide progenitors, compared to 25% embryonic (hESCs)....
Leber's Hereditary Optic Neuropathy (LHON) is a maternally inherited disorder caused by homoplasmic mutations of mitochondrial DNA (mtDNA). LHON characterized the selective degeneration retinal ganglion cells (RGC). Almost all maternal lineages are mutant (100% mtDNA copies mutant) for one three frequent now found in over 90% patients worldwide (m.11778G > A/MT-ND4, m.3460G A/MT-ND1, m.14484 T C/MT-ND6). Human induced pluripotent stem (hiPSCs) were generated from patient carrying A/MT-ND1...
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 DNA (mtDNA), with key being m.3460G > A, m.11778G m.14484 T C. Fibroblasts identical twins, sharing C m.10680G A variants each 70 % heteroplasmy, were used to generate iPSC lines. Remarkably, twin, a LHON patient, displayed symptoms, while other, carrier, remained asymptomatic. These iPSCs offer valuable...
The generation of inducible pluripotent stem cells (iPSCs) is a revolutionary technique allowing production patient-specific cell lines used for disease modeling, drug screening, and therapy. Integrity nuclear DNA (nDNA) mandatory to allow iPSCs utilization, while quality control mitochondrial (mtDNA) rarely included in the validation process. In this study, we performed mtDNA deep sequencing during transition from parental fibroblasts reprogrammed iPSC differentiated neuronal precursor...
More than 30 years after discovering Leber's hereditary optic neuropathy (LHON) as the first maternally inherited disease associated with homoplasmic mtDNA mutations, we still struggle to achieve effective therapies. LHON is characterized by selective degeneration of retinal ganglion cells (RGCs) and most frequent mitochondrial disease, which leads young people blindness, in particular males. Despite that causative mutations are present all tissues, only a specific cell type affected. Our...
Pearson marrow pancreas syndrome (PMPS) is a sporadic mitochondrial disease, resulting from the clonal expansion of mutated DNA (mtDNA) molecule bearing macro-deletion, and therefore missing essential genetic information. PMPS characterized by presence deleted (Δ) mtDNA that co-exist with variable amount wild-type mtDNA, condition termed heteroplasmy. All tissues affected individual, including haemopoietic system post-mitotic, highly specialized (brain, skeletal muscle, heart) contain...
Summary Adeno-Associated Virus 9 (AAV9) is a delivery platform highly exploited to develop gene-based treatments for neurological disorders given its low pathogenicity and brain tissue tropism. However, the efficacy of this vector dampened by relatively efficiency cross adult blood-brain barrier (BBB) inherent targeting liver upon intravenous delivery. We generated new peptide display library starting from galactose binding-deficient AAV9 capsid selected two engineered capsids, named AAV-Se1...
Abstract Rett syndrome (RTT) is an incurable neurodevelopmental disorder caused by mutations in the gene encoding for methyl-CpG binding-protein 2 (MeCP2). Gene therapy this disease presents inherent hurdles since MECP2 expressed throughout brain and its duplication leads to severe neurological conditions as well. However, recent introduction of AAV-PHP.eB, engineered capsid with unprecedented efficiency crossing blood-brain barrier upon intravenous injection, has provided invaluable vehicle...
Summary Wolfram syndrome 1 (WS1) is a rare genetic disorder caused by mutations in the WFS1 gene leading to wide spectrum of clinical dysfunctions, among which blindness, diabetes and neurological deficits are most prominent. encodes for endoplasmic reticulum (ER) resident transmembrane protein Wolframin with multiple functions ER processes. However, -dependent etiopathology retinal cells unknown. Herein, we showed that Wfs1 mutant mice developed early electrophysiological impairments...