Research lines

Unravelling the role of mitochondrial (dys)function in AD pathogenesis

We use state-of-the-art technologies and tools for a wide array of analyses, both in vitro and in vivo. Our research covers mitochondrial Ca²⁺ handling, organelle coupling with other intracellular structures, and organelle metabolism/bioenergetics at different disease stages in PS2-based AD mouse models. We are also interested in verifying possible links between mitochondrial defects and behavioural impairment.

Relevant publications

1. Rossi A, Rigotto G, Valente G, Giorgio V, Basso E, Filadi R, Pizzo P. (2020) Defective mitochondrial pyruvate flux affects cell bioenergetics in Alzheimer's Disease-related models. Cell Reports 30:2332‑2348.e10
2. Theurey P, Connolly NMC, Fortunati I, Basso E, Lauwen S, Ferrante C, Moreira Pinho C, Joselin A, Gioran A, Bano D, Park DS, Ankarcrona M, Pizzo P, Prehn JHM. (2019) Systems biology identifies preserved integrity but impaired metabolism of mitochondria due to a glycolytic defect in Alzheimer's disease neurons. Aging Cell 18:e12924
3. Filadi R, Greotti E, Turacchio G, Luini A, Pozzan T, Pizzo P. (2016) Presenilin 2 modulates endoplasmic reticulum-mitochondria coupling by tuning the antagonistic effect of Mitofusin 2. Cell Reports 15:2226‑2238
4. Zampese E, Fasolato C, Kipanyula MJ, Bortolozzi M, Pozzan T, Pizzo P (2011) Presenilin 2 modulates endoplasmic reticulum (ER)-mitochondria interactions and Ca²⁺ cross-talk. Proc Natl Acad Sci USA, 108:2777–2782

Ca²⁺ homeostasis defects in AD models

Cytosolic and organelle Ca²⁺ handling is studied in AD cell and animal models, not only in neurons but also in glial cells, both in vitro and at the circuit level. The role of amyloid accumulation and defective Ca²⁺ homeostasis on brain functionality is also investigated by recording spontaneous brain oscillatory activity of AD mice in vivo.

Relevant publications

1. Pizzo P, Basso E, Filadi R, Greotti E, Leparulo A, Pendin D, Redolfi N, Rossini M., Vajente N, Pozzan T. and Fasolato C. “Presenilin-2 and Calcium Handling: Molecules, Organelles, Cells and Brain Networks” Cells 2020, 9(10) 2166; https://doi.org/10.3390/cells9102166 PMID: 32992
2. Scremin E, Agostini M, Leparulo A, Pozzan T, Greotti E, Fasolato C. ORAI2 Down-Regulation Potentiates SOCE and Decreases Aβ42 Accumulation in Human Neuroglioma Cells. Int J Mol Sci. 2020 Jul 25;21(15):5288. doi:10.3390/ijms21155288. PMID: 32722509; PMCID: PMC7432374
3. Leparulo A, Mahmud M, Scremin E, Pozzan T, Vassanelli S, Fasolato C. Dampened Slow Oscillation Connectivity Anticipates Amyloid Deposition in the PS2APP Mouse Model of Alzheimer's Disease.” Cells 2020, 9(1) pii: E54. doi: 10.3390/cells9010054 (2020) 9(1). PMID: 31878336
4. Greotti E, Capitanio P, Wong A, Pozzan T, Pizzo P, Pendin D. (2019). Familial Alzheimer’s disease-linked presenilin mutants and intracellular Ca²⁺ handling: A single-organelle, FRET-based analysis. Cell Calcium, 79:44–56.

Organelle coupling in physiology and disease

We investigate the significance of membrane contact sites, both in cell physiology and pathology, with a special focus on interorganelle Ca²⁺ signalling. A major interest is the development of novel tools to study organelle coupling.

Relevant publications

1. Filadi R, De Mario A, Audano M, Romani P, Pedretti S, Cardenas C, Dupont S, Mammucari C, Mitro N, Pizzo P (2023) "Sustained IP3-linked Ca²⁺ signaling promotes progression of triple negative breast cancer cells by regulating fatty acid metabolism" Front. Cell Dev. Biol. 11:1071037. PMID: 36994106.
2. Rossini M, García-Casas P, Filadi R, Pizzo P (2021) "Loosening ER–mitochondria coupling by the expression of the presenilin 2 loop domain" Cells 10:1968. PMID: 34440738.
3. Filadi R, Leal NS, Schreiner B, Rossi A, Dentoni G, Pinho CM, Wiehager B, Cieri D, Calì T, Pizzo P, Ankarcrona M (2018) "TOM70 sustains cell bioenergetics by promoting IP3R3-mediated ER to mitochondria Ca²⁺ transfer" Curr Biol. 28:369-382.e6. PMID: 29395920.
4. Filadi R, Greotti E, Turacchio G, Luini A, Pozzan T, Pizzo P (2016) "Presenilin 2 modulates endoplasmic reticulum-mitochondria coupling by tuning the antagonistic effect of mitofusin 2" Cell Rep. 15:2226-2238. PMID: 27239030.
5. Filadi R, Greotti E, Turacchio G, Luini A, Pozzan T, Pizzo P (2015) "Mitofusin 2 ablation increases endoplasmic reticulum-mitochondria coupling" Proc. Natl. Acad. Sci. USA 112:E2174-E2181. PMID: 25870285.

Modeling neurodegenerative disorders in Drosophila melanogaster

We use Drosophila as a model to study the neurodegenerative disorder Hereditary Spastic Paraplegia. We are focused on understanding how ER shape-impairment, due to HSP gene mutations, affects specific ER functions.

Relevant publications

1. Montagna A, Vajente N, Pendin D, Daga A. In vivo Analysis of CRISPR/Cas9 Induced Atlastin Pathological Mutations in Drosophila. Front Neurosci. 2020 Oct 15;14:547746
2. Vajente N, Norante R, Redolfi N, Daga A, Pizzo P, Pendin D. Microtubules Stabilization by Mutant Spastin Affects ER Morphology and Ca²⁺ Handling. Front Physiol. 2019 Dec 20;10:1544

Subcellular indicators

We are interested in the generation of chemical and genetically-encoded biosensors, for both Ca²⁺ and other ions and metabolites, targeted to specific subcellular locations to finely dissect intracellular signalling pathways. These tools are instrumental for determining a high-resolution map of the heterogeneity of signalling pathways within the cell. Furthermore, we develop tools to manipulate the functions of organelles, exploiting state-of-the-art methods, such as optogenetics.

Relevant publications

1. Greotti E, Fortunati I, Pendin D, Ferrante C, Galla L, Zentilin L, Giacca M, Kaludercic N, Di Sante M, Mariotti L, Lia A, Gómez-Gonzalo M, Sessolo M, Di Lisa F, Carmignoto G, Bozio R, Pozzan T. mCerulean3-Based Cameleon Sensor to Explore Mitochondrial Ca2+ Dynamics In Vivo. iScience. 2019 Jun 28;16:340-355. doi: 10.1016/j.isci.2019.05.031. Epub 2019 May 27. Erratum in: iScience. 2019 Sep 27;19:161. PMID: 31203189; PMCID: PMC6581653
2. Tkatch T, Greotti E, Baranauskas G, Pendin D, Roy S, Nita LI, Wettmarshausen J, Prigge M, Yizhar O, Shirihai OS, Fishman D, Hershfinkel M, Fleidervish IA, Perocchi F, Pozzan T, Sekler I. Optogenetic control of mitochondrial metabolism and Ca²⁺ signaling by mitochondria-targeted opsins. Proc Natl Acad Sci U S A. 2017 Jun 27;114(26):E5167-E5176. doi: 10.1073/pnas.1703623114. Epub 2017 Jun 13. PMID: 28611221; PMCID: PMC5495261
3. Greotti E, Wong A, Pozzan T, Pendin D, Pizzo P. Characterization of the ER-Targeted Low Affinity Ca²⁺ Probe D4ER. Sensors (Basel). 2016 Sep 2;16(9):1419. doi: 10.3390/s16091419. PMID: 27598166; PMCID: PMC5038697
4. Pendin D, Norante R, De Nadai A, Gherardi G, Vajente N, Basso E, Kaludercic N, Mammucari C, Paradisi C, Pozzan T, Mattarei A. A Synthetic Fluorescent Mitochondria-Targeted Sensor for Ratiometric Imaging of Calcium in Live Cells. Angew Chem Int Ed Engl. 2019 Jul 15;58(29):9917-9922

Active grants

• Chan Zuckerberg Initiative • Frontiers of Imaging Grant • Grant 2023-321185 • Dynamic chemogenetic reporters to decipher membrane contacts • 2023-2026.
• Progetti di Ricerca di Rilevante Interesse Nazionale (PRIN 2022; Italian Ministry of Research Grant 202282LYW5) • Innovative chemical tools for real-time calcium imaging in organelles • 2023-2025.
• Progetti di Ricerca di Rilevante Interesse Nazionale (PRIN 2022; Italian Ministry of Research Grant 20222943TH9) • Purinergic checkpoints in neuroinflammation and Alzheimer's disease: extracellular ATP and the P2X7 receptor as main drivers of neurodegeneration • 2023-2025.
• Fondazione Cariparo and Fondazione Telethon • Grant GJC21054 • Deorphanizing and functionalizing the mitochondrial protein TMEM65 • 2022-2024.
• Cure Alzheimer's Fund • Extracellular ATP is a key factor in promoting Alzheimer's disease neuroinflammation • 2021-2023 + Follow-on Project 2024-2025

Our lab is also part of the Advanced Light Microscopy Italian Node of Euro-BioImaging.