Laboratory of Applied Biology
Our laboratory is involved in different main projects:
1) the study of the molecular mechanisms at the basis of Amyotrophic Lateral Sclerosis (ALS) and of SpinoBulbar Muscular Atrophy (SBMA), two different motor neuron diseases (MNDs) characterized by similar pathogenic mechanisms.
2) the role of sex steroids on stress tolerance in hormone-dependent cancers.
Line 1) With regards to MNDs (ALS and SBMA) we are studying how aberrant mutant conformations (misfolded species) of disease associated proteins aggregate and become neurotoxic to neuron and muscle cells. Our main goal is to prevent their misfolding and accumulation to counteract motoneuronal loss and muscle damages. In this context, in this MNDs we identified the protective effects of a novel form of chaperone-assisted selective autophagy (CASA) and its interplay with the proteasome system. We found compounds that are able to potentiate CASA and the routing of misfolded proteins for their degradation which may constitute novel potential therapeutic approaches for these fatal disorders.
Line 2) In the project we are focused on how steroids hormones may modulate stress tolerance in prostate and breast cancer cells, allowing to them to proliferate and migrate even when they are exposed to chemiotherapics. Mediator of the increased cell stress tolerance are the small heat shock protein B8 and its partner BAG3, a co-chaperone of HSP70. Together, these proteins protect cancer cells from stress-induced cell death.
Several techniques are in use in our laboratory, ranging from DNA manipulation, plasmid construction, cell culture and transfection, basis molecular and cell biology techniques, such as immunofluorescence and confocal microscopy, gene expression (real time-PCR) and protein detection (western blot, Elisa assay, etc.), aggregation assay (filter retardation assay, FLOiT assay), evaluation of the degradative systems (proteasome and autophagy assays, lysosome integrity analysis, etc.), flow cytometry, etc.
Several different cell lines are in use (immortalized motoneuronal cells, stabilized myoblasts, neuroblastoma cells, prostate and breast cancer cells) for routine experiments in transient or stable transfections.
Patient derived iPSCs and their corresponding isogenic cells lines (obtained with CRISPR/Cas9 technology) are available. iPSCs differentiation to neuronal/motoneuronal cells is routinely performed.
Several collaborations are in progress supported by Italian and European grants. In particular, the most active collaboration in Italy are in progress with the IRCCS Pharmacology Inst. M. Negri, Milano, the IRCCS Neurological Ist C. Besta, Milano, the IRCCS Neurological Ist. C. Mondino, Pavia, and the Universities of Milano-Bicocca, of Modena, of Trento, of Padua, of Genoa, of Turin. European collaboration is in progress with the Max Plank Inst of Dresden, the Technische Universität Dresden DFG-Center for Regenerative Therapies Dresden, The University of Oxford, the University College of London, the University of Antwerpen, Belgium. We also have collaboration with the University of Michigan, the Baylor College of Medicine, Houston Tx, USA.
1: Rusmini P, Cortese K, Crippa V, Cristofani R, Cicardi ME, Ferrari V, Vezzoli G, Tedesco B, Meroni M, Messi E, Piccolella M, Galbiati M, Garrè M, Morelli E, Vaccari T, Poletti A.
Trehalose induces autophagy via lysosomal-mediated TFEB activation in models of motoneuron degeneration. Autophagy. 2018 Oct 18:1-21.
doi: 10.1080/15548627.2018.1535292. [Epub ahead of print] PubMed PMID: 30335591.
2: Cristofani R, Crippa V, Rusmini P, Cicardi ME, Meroni M, Licata NV, Sala G, Giorgetti E, Grunseich C, Galbiati M, Piccolella M, Messi E, Ferrarese C, Carra S, Poletti A.
Inhibition of retrograde transport modulates misfolded protein accumulation and clearance in motoneuron diseases. Autophagy. 2017 Aug 3;13(8):1280-1303.
doi: 10.1080/15548627.2017.1308985. PubMed PMID: 28402699; PubMed Central PMCID: PMC5584856.
3: Crippa V, D'Agostino VG, Cristofani R, Rusmini P, Cicardi ME, Messi E, Loffredo R, Pancher M, Piccolella M, Galbiati M, Meroni M, Cereda C, Carra S, Provenzani A, Poletti A.
Transcriptional induction of the heat shock protein B8 mediates the clearance of misfolded proteins responsible for motor neuron diseases.
Sci Rep. 2016 Mar 10;6:22827. doi: 10.1038/srep22827. PubMed PMID: 26961006; PubMed Central PMCID: PMC4785366.
4: Giorgetti E, Rusmini P, Crippa V, Cristofani R, Boncoraglio A, Cicardi ME, Galbiati M, Poletti A.
Synergic prodegradative activity of Bicalutamide and trehalose on the mutant androgen receptor responsible for spinal and bulbar muscular atrophy.
Hum Mol Genet. 2015 Jan 1;24(1):64-75.
doi: 10.1093/hmg/ddu419. Epub 2014 Aug 13. PubMed PMID: 25122660; PubMed Central PMCID: PMC4262493.
5: Cristofani R, Montagnani Marelli M, Cicardi ME, Fontana F, Marzagalli M, Limonta P, Poletti A, Moretti RM.
Dual role of autophagy on docetaxel-sensitivity in prostate cancer cells.
Cell Death Dis. 2018 Aug 30;9(9):889.
doi: 10.1038/s41419-018-0866-5. PubMed PMID: 30166521; PubMed Central PMCID: PMC6117300.