RESEARCH INTERESTS: Cellular and molecular mechanisms of striated muscle physiopathology

1. PHARMACOLOGICAL, PHYSICAL, AND NUTRITIONAL INTERVENTIONS AGANIST CANCER CACHEXIA: My laboratory is focused on different approaches to counteract cancer cachexia, including pharmacological (exercise mimetics), physiological (physical activity), and nutritional (supplements) interventions in humans and animal models. 2. MYOFIBER MEMBRANE DAMAGE AND REPAIR: Duchenne Muscular Dystrophy (DMD), is a lethal genetic, muscle-wasting disease, characterized by progressive muscle fragility and weakness. The muscle membrane repair mechanism (MRM) is an active resealing pathway involving vesicle-sarcolem fusion to “patch” the compromised plasma membrane and represents a possible target to counteract muscle wasting in DMD, in which the chronic cycle of muscle degeneration-regeneration plays a pivotal role in disease progression. 3. PATENTS AND TECHNOLOGY TRASNFER: I am co-inventor of a patented procedure to produce Hsp60-enriched exosomes with exercise-mimetic activity, a product that is, therefore, called Physiactisome. Patent: Physiactisome – «Procedure for the synthesis of HSP-containing exosomes and their use against muscle atrophy and cachexia» - patent n. 102018000009235 on 8/10/2018, deposited by Università di Palermo. Owners: Università di Palermo, Università di Roma La Sapienza, Nanovector Torino, Sorbonne Université. List of inventors: Valentina Di Felice, Rosario Barone, Antonella Marino Gammazza, Campanella Claudia, Cappello Francesco, Farina Felicia, Eleonora Trovato, Daniela D’Amico, Filippo Macaluso, Dario Coletti, Sergio Adamo, Gabriele Multhoff, Paolo Gasco. International publication number WO 2020/075004 A1. This product can be exploited against muscle atrophy, since it ameliorates muscle endurance and homeostasis. The presentation of the product and the corresponding Spinoff project (iBioTHEx) was awarded the third prize at the EIT JumpStarter Grand final, Riga, Latvia, 15-17/11/2019, Health category. 4. PHYSIOPATHOLOGY OF MUSCLE TISSUES: I contribute to discovering and explaining those mechanisms underlying pathologies of the striated and smooth muscle tissues; this activity is carried out at Sorbonne University by using genetic murine models.

Cancer cachexia

Cancer cachexia
Compared to a control mouse (left) a tumor-bearing mouse (right) displays a dramatic muscle wasting. This loss of muscle mass is called cancer cachexia.

Exogenous gene expression in regenerating muscle

Exogenous gene expression in regenerating muscle
Depicted here is the over-expression of Green Fluorescent Protein (GFP, green; click on the image to access Tsien's Lab) in interstitial cells (circled), nascent myofibers (arrow) and adult fibers (arrowhead), in a regenerating Tibialis Anterior following focal injury. Laminin staining (red) highlights the basement membrane surrounding the skeletal muscle tissue, while nuclei are stained in blue. We do gene delivery by electroporation to study the regulation of muscle regeneration.

RESEARCH INTERESTS: Tissue engineering of skeletal muscle

Background and rationale.

Tissue engineering lies at the interface of regenerative medicine and developmental biology, and represent an innovative and multidisciplinary approach to build organs and tissues (Ingber and Levin, Development 2007). The skeletal muscle is a contractile tissue characterized by highly oriented bundles of giant syncytial cells (myofibers) and by mechanical resistance. Contractile, tissue-engineered skeletal muscle would be of significant benefit to patients with muscle deficits secondary to congenital anomalies, trauma, or surgery. Obvious limitations to this approach are the complexity of the musculature, composed of multiple tissues intimately intermingled and functionally interconnected, and the big dimensions of the majority of the muscles, which imply the involvement of an enormous amount of cells and rises problems of cell growth and survival (nutrition and oxygen delivery etc.). Two major approaches are followed to address these issues. Self-assembled skeletal muscle constructs are produced in vitro by delaminating sheets of cocultured myoblasts and fibroblasts, which results in contractile cylindrical “myooids.” Matrix-based approaches include placing cells into compacted lattices, seeding cells onto degradable polyglycolic acid sponges, seeding cells onto acellularized whole muscles, seeding cells into hydrogels, and seeding nonbiodegradable fiber sheets. Recently, decellularized matrix from cadaveric organs has been proven to be a good scaffold for cell repopulation to generate functional hearts in mice (Ott et al. Nature Medicine 2008).

I have obtained cultures of skeletal muscle cells on conductive surfaces, which is required to develop electronic device–muscle junctions for tissue engineering and medical applications1. I aim to exploit this system for either recording or stimulation of muscle cell biological activities, by exploiting the field effect transistor and capacitor potential of the conductive substratum-cell interface. Also, we are able to create patterned dispositions of molecules and cells on gold, which is important to mimic the highly oriented pattern myofibers show in vivo.

I have found that Static magnetic fields enhance skeletal muscle differentiation in vitro by improving myoblast alignment2. Static magnetic field (SMF) interacts with mammal skeletal muscle; however, SMF effects on skeletal muscle cells are poorly investigated. 80 +/- mT SMF generated by a custom-made magnet promotes myogenic cell differentiation and hypertrophy in vitro. Finally, we have transplanted acellular scaffolds to study the in vivo response to this biomaterial3, which we want to exploit for tissue culture and regenerative medicine of skeletal muscle.

The specific aims of my current research are:

1) to increase and optimize the production and alignment of myogenic cells and myotubes in vitro;

2) to manipulate the niche of muscle stem cells aimed at ameliorating their regenerative capacity in vivo;

3) to develop muscle-electrical devices interactions. We plan to exploit the cell culture system on conductive substrates for either recording or stimulation of muscle cell biological activities, by exploiting the field effect transistor and capacitor potential of the conductive substratum-cell interface.

4) to better clarify the biological effects of Static Magnetic Fields. With the aim to characterize the molecular mechanism underlying the effects of SMF on cell differentiation and alignment we are exposing molecules and cells to SMF below 1T.
5) to produce pre-assembled, off-the-shelf skeletal muscle. We are seeding acellularized muscle scaffold with various cell types, with the goal to obtain functional muscle with vascular supply and nerves.


REFERENCES

1) Coletti D. et al., J Biomed Mat Res 2009; 91(2):370-377.

2) Coletti D. et al., Cytometry A. 2007;71(10):846-56.


3) Perniconi B. et al. Biomaterials, 2011 in press

Cultures of myotubes on a conductive surface in a parallel orientation.

Cultures of myotubes on a conductive surface in a parallel orientation.
C2C12 cells cultured on gold, by mean of adhesion to 100 nm-wide stripes coated with anti Stem Cell antigen1 (Sca1) Ab. Nuclei (blue) and actin cytoskeleton (red) staining highlights the selective cells adhesion on the Ab-coated stripes and the formation of parallel multinucleated syncytia (myotubes).

6/20/2011

Blind tasting session at the lab


To celebrate a few recent events (the UPMC Emergence 2011 grant, the Mol Endocrinol paper) and to welcome a new student in the lab, we have tasted five Bordeaux 2006 wines, from different appellations characterized by marked nuances of their terroirs and specific grape assembly. Given that the different wineyards are only about 50 Km from each other, the differences were outstanding.

Results of the blind tasting (panel : laboratory members):

1st Château-Haut Maurac, Médoc Cru Bourgeois (60 % Cabernet sauvignon, 40 % Merlot)
2nd Château Musset Chevalier , Saint Emillon Grand cru (50 % Merlot noir / 45 % Cabernet-Franc / 5 % Cabernet-Sauvignon )
3rd Les Hauts du Tertre, Margaux (55 % Cabernet sauvignon, 20 % Merlot, 20 % Cabernet franc, 5 % Petit verdot)
4th Château Prieuré-les-Tours, Graves.

We liked the winner for its intense bouquet of red fruits and its full body, with mature tannins and a long lasting aftertaste. One more cru Borgeois showing the great quality/price ratio of this category. From the color to the marked tannins it expressed the Medoc pretty well. However, I preferred the Margaux of Les Hauts de Tertre, a second wine produced by Château du Tertre, for its elegance and its more floreal bouquet. Margaux came out in the good balance between tannins, acidity and alcoolic warmth. The superb roundness of the Libournais St Emillon and the acidity of the Graves (Alas! - in such a poor interpretation) came out as well, but nobody guessed the crus for all the wines.
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ARTICLES: Toschi et al. Mol Endocrinol 2011



In this paper, entitled "SKELETAL MUSCLE REGENERATION IN MICE IS STIMULATED BY LOCAL OVEREXPRESSION OF
V1a-VASOPRESSIN RECEPTOR", we identify skeletal muscle as a physiological target of hormones of the vasopressin (AVP) family and show a novel in vivo role for vasopressin-dependent pathways. FIG LEGEND In red Myc (i.e. overexpressed V1a-R) immunolocalization in skeletal muscle fibers highlighted by laminin staining in green.

In the last 10 years, we have characterized in detail AVP signaling pathways in myogenic cells in vitro. Also, we have reported that the muscle specific, V1a, AVP-receptor is modulated during myogenic differentiation in vivo, which suggest a role in muscle development. Consistently, we have shown that AVP intramuscular injection enhances muscle regeneration, a process which recapitulates muscle development in the adult.
With the last paper by Toschi et al. we formally demonstrate the biological role of AVP on skeletal muscle homeostasis and we pinpoint some molecular mechanisms underlying this effect, including calcineurin-mediated IL-4 production in the musculature in response to AVP.

FIG LEGEND role of Calcineurin-dependent effects of V1a-R overexpression on muscle regeneration. Further links to the press which cited the article: ANSA and Corriere della Sera
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Against cuts in cultural funding

Again a non scientific, still relevant post insomuch as politics affect culture, research and education.

Cuts on culture and arts.
We have recently celebrated the 150th anniversary of Italian unification. As reported by the New York Times, a very intense moment occurred when Riccardo Muti conducted the "Va pensiero" at the premiere of Verdi's “Nabucco” at the Teatro dell’Opera in Rome in March, in the presence of the Prime Minister and the Mayor of the capital.
The issue was the heavy cut plan on cultural founding performed by the current government. The event had its climax at Muti's brief statements against this plan while introducing an exceptional bis of the "Va pensiero". Linked to the title of this post there is the touching video on youtube.

Cuts and management of university funding.
University budget cuts represent the other branch of the current harmful intervention on state budget, in a country which spending on university is already very low as compared to most other countries, as reported by the BBC last year. However, it is not only a matter of budget. What is even worse is the total incertitude for the CURRENT available fundings: in 2011 we are still waiting for the results of a major funding call of the Italian Ministry for the University and Research (MIUR) which is called PRIN 2009 and was released in 2010! In 2012, if and when some groups will receive the grants to which they applied three years before, what will remain to be accomplished of the proposed research projects? Won't the latter be born already aged and out to date?
A lucid analysis on the inceritude which reigns on italian university has been published a few months ago on the web pages of lavoce.info (in Italian).
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THE NETWORK OF OUR COLLABORATORS 2017

THE NETWORK OF OUR COLLABORATORS 2017
We collaborate with the Myology Group and the Cochin Hospital in Paris for stem cell studies and SRF, with the Cancer Centre at Ohio State University, Columbus for studies on the mechanisms underlying cachexia, with the Neurorehabilitation Unit at University of Pisa for clinical studies, with Pharmacology and Bioinformatics at the University of Urbino for advanced statistical analyses, with the Anatomy Section at the University of Perugia and with GYN/OB at the University of Western Piedmont for studies related to circulating factors and myogenic cell responses in cachexia, with the Biotech-Med Unit at ENEA, Chemistry in Rome and Anatomy in palermo for tissue engineering applications. Functional studies are carried out in our Departement in Rome in collaboration with Musaro's laboratory.