San Raffaele Telethon Institute for Gene Therapy
Gene and neural stem cell therapy for lysosomal storage diseases
The overarching goal of the lab is to study and treat rare genetic diseases affecting the CNS, with a special focus on Lysosomal Storage Disorders (LSDs), including leukodystrophies - Metachromatic leukodystrophy (MLD), Globoid-cell Leukodystrophy (GLD) and Alexander's disease (AxD) - and GM2 gangliosidosis - Sandhoff Disease (SD) and Tay-Sachs Disease (TSD). Our projects span from basic to translational research, using murine and human models, and testing different therapeutic platforms (i.e. in vivo GT, ex vivo GT; cell therapy, gene addition, gene editing).
Research activity
Our active projects fall within three research areas:
- Enhance gene therapy strategies for LSDs. Over the past 10 years we have shown the safety and efficacy of intracerebral LV GT in supplying therapeutic levels of functional lysosomal enzymes to ameliorate/rescue the pathology in murine and NHP models of LSDs. The main goals for this area will be: i) to move forward the clinical testing of intracerebral LV GT in MLD patients; ii) to enhance the efficacy of GT strategies, coupling enzyme overexpression achieved in effector cells with enhanced transgene bioavailability and more efficient metabolic correction of enzyme-deficient cells
- Modelling LSDs using iPSC-based systems. The lab has optimized culture conditions, transduction and neural differentiation protocols of human iPSCs derived from LSD patients (and non-affected controls). We have used these cells to test cell/gene therapy approaches and to model diseases, as patient-specific iPSC-derived neural populations recapitulate key biochemical features of the disease and allow for the identification of early defects in human neuronal/glial cell populations. Our goal in this area is to exploit and adapt/optimize these models to generate hiPSC-based platforms that are relevant in the context of LSDs by developing: i) co-cultures obtained by mixing defined neuronal and/or glial cell population; 3D systems (brain organoids generated using different protocols).
- Developing gene editing strategies to treat CNS disease. This area started less than two years ago under the supervision of Dr. V. Meneghini (Project Leader) and benefits from our unique expertise in CNS biology/disease to implement state-of-the art gene and base editing strategies that are being extensively tested in other tissues (i.e. liver, hematopoietic system) at SR-Tiget and worldwide. Our initial focus is Alexander's Disease, a rare autosomal dominant leukodystrophy caused by missense mutations in the gene encoding the glial fibrillary acidic protein (GFAP), the major intermediate filament protein in astrocytes. We aim to provide in vivo proof-of-concept of safety, efficiency and efficacy of a novel approach based on gene editing/base editing strategies targeting GFAP mutational hotspots to correct pathological phenotypes in astrocytes. If successful, these new editing platforms for in vitro and in vivo targeting of the CNS could be prospectively applied for disease modeling and treatment of other glial and neurodegenerative diseases.
Sala D, Ornaghi F, Morena F, Argentati C, Valsecchi M, Alberizzi V, Di Guardo R, Bolino A, Aureli M, Martino S, Gritti A. Therapeutic advantages of combined gene/cell therapy strategies in a murine model of GM2 gangliosidosis. Mol Ther Methods Clin Dev. 2022 Jun 9;25:170-189.
Giordano AMS, Luciani M, Gatto F, Abou Alezz M, Beghè C, Della Volpe L, Migliara A, Valsoni S, Genua M, Dzieciatkowska M, Frati G, Tahraoui-Bories J, Giliani SC, Orcesi S, Fazzi E, Ostuni R, D'Alessandro A, Di Micco R, Merelli I, Lombardo A, Reijns MAM, Gromak N, Gritti A, Kajaste-Rudnitski A. DNA damage contributes to neurotoxic inflammation in Aicardi-Goutières syndrome astrocytes. J Exp Med. 2022 Apr 4;219(4).
Mangiameli E, Cecchele A, Morena F, Sanvito F, Matafora V, Cattaneo A, Della Volpe L, Gnani D, Paulis M, Susani L, Martino S, Di Micco R, Bachi A & Gritti A (2021) Human iPSC-based neurodevelopmental models of globoid cell leukodystrophy uncover patient- and cell type-specific disease phenotypes. Stem Cell Reports 16: 1–18
Meneghini V, Peviani M, Luciani M, Zambonini G & Gritti A (2021) Delivery Platforms for CRISPR/Cas9 Genome Editing of Glial Cells in the Central Nervous System. Front. Genome Ed. 3: 644319
Ricca A, Cascino F, Morena F, Martino S, Gritti A. In vitro Validation of Chimeric β-Galactosylceramidase Enzymes With Improved Enzymatic Activity and Increased Secretion. Front Mol Biosci. 2020 Jul 21;7:167. doi: 10.3389/fmolb.2020.00167. eCollection 2020. PMID: 32850960 Free PMC article.
Ricca A, Rufo N, Ungari S, Morena F, Martino S, Kulik W, Alberizzi V, Bolino A, Bianchi F, Del Carro U, Bi A & Gritti A, Combined gene/cell therapies provide long-term and pervasive rescue of multiple pathological symptoms in a murine model of globoid cell leukodystrophy. Hum. Mol. Genet., 2015, 24(12): 3372-89.
Meneghini V, Lattanzi A, Tiradani L, Bravo G, Morena F, Sanvito F, Calabria A, Bringas J, Fisher-Perkins JM, Dufour JP, Baker KC, Doglioni C, Montini E, Bunnell BA, Bankiewicz K, Martino S, Naldini L & Gritti A, Pervasive supply of therapeutic lysosomal enzymes in the CNS of normal and Krabbe-affected non-human primates by intracerebral lentiviral gene therapy. EMBO Mol. Med. 2016, 8: 489–510.
Meneghini V, Frati G, Sala D, De Cicco S, Luciani M, Cavazzin C, Paulis M, Mentzen W, Morena F, Giannelli S, Sanvito F, Villa A, Bulfone A, Broccoli V, Martino S & Gritti A, Generation of Human Induced Pluripotent Stem Cell-Derived Bona Fide Neural Stem Cells for Ex Vivo Gene Therapy of Metachromatic Leukodystrophy. Stem Cells Transl. Med., 2016, 6: 352–368.
Mazzara PG, Massimino L, Pellegatta M, Ronchi G, Ricca A, Iannielli A, Giannelli SG, Cursi M, Cancellieri C, Sessa A, Del Carro U, Quattrini A, Geuna S, Gritti A, Taveggia C & Broccoli V, Two factor-based reprogramming of rodent and human broblasts into Schwann cells. Nat. Commun. 2017, 8:14088
Frati G, Luciani M, Meneghini V, De Cicco S, Ståhlman M, Blomqvist M, Grossi S, Filocamo M, Morena F, Menegon A, Martino S & Gritti A, Human iPSC-based models highlight defective glial and neuronal differentiation from neural progenitor cells in metachromatic leukodystrophy. Cell Death Dis. 2018, 9: 698.
Ornaghi F, Sala D, Tedeschi F, Ma a MC, Bazzucchi M, Morena F, Valsecchi M, Aureli M, Martino S, Gritti A. Novel bicistronic lentiviral vectors correct β-Hexosaminidase de ciency in neural and hematopoietic stem cells and progeny: implications for in vivo and ex vivo gene therapy of GM2 gangliosidosis. Neurobiol Dis. 2019 Nov 1;134:104667.
Frati G, Luciani M, Meneghini V, De Cicco S, Ståhlman M, Blomqvist M, Grossi S, Filocamo M, Morena F, Menegon A, Martino S, Gritti A. Human iPSC-based models highlight defective glial and neuronal differentiation from neural progenitor cells in metachromatic leukodystrophy. Cell Death Dis. 2018 Jun 13;9(6):698
Meneghini V, Frati G, Sala D, De Cicco S, Luciani M, Cavazzin C, Paulis M, Mentzen W, Morena F, Giannelli S, Sanvito F, Villa A, Bulfone A, Broccoli V, Martino S, Gritti A. Generation of human induced pluripotent stem cell-derived bona fide neural stem cells for ex vivo gene therapy of metachromatic leukodystrophy. Stem Cells Trasl Med. 2017 Feb;6(2):352-368.
Meneghini V, Lattanzi A, Tiradani L, Bravo G, Morena F, Sanvito F, Calabria A, Bringas J, Fisher-Perkins JM, Dufour JP, Baker KC, Doglioni C, Montini E, Bunnell BA, Bankiewicz K, Martino S, Naldini L, Gritti A. Pervasive supply of therapeutic lysosomal enzymes in the CNS of normal and Krabbe-affected non-human primates by intracerebral lentiviral gene therapy. EMBO Mol Med. 2016 May 2;8(5):489-510.
Ricca A, Rufo N, Ungari S, Morena F, Martino S, Kulik W, Alberizzi V, Bolino A, Bianchi F, Del Carro U, Biffi A, Gritti A. Combined gene/cell therapies provide long-term and pervasive rescue of multiple pathological symptoms in a murine model of globoid cell leukodystrophy. Hum Mol Genet. 2015 Jun 15;24(12):3372-89.