Marco A. Cassatella, M.D.

Professor of General Pathology

cassatella

 

Department of Pathology and Diagnostics
Division of General Pathology
University of Verona, Faculty of Medicine

Strada Le Grazie 4 37134 Verona, Italy

Office: ++ 39 (045)8027130
Lab: ++ 39 (045)8027259
FAX: ++ 39 (045)8027127

email: marco.cassatella@univr.it

 

 

Marco A. Cassatella specialized in Clinical Biology on October 31, 1988 at the University of Verona, year in which he was appointed Research Assistant. He became Associate Professor in 1992 and Professor of General Pathology at the Medical School of the University of Verona in 2001. Since 1983 he has carried on research and teaching as a scolarship holder of the "Consorzio per lo Sviluppo degli Studi Universitari" at the Institute of the General Pathology of the Medical School of the University of Verona. Supported by a fellowship of the "Italian Association for Cancer Research", he spent two years, from January 19, 1987, to February 21, 1989, at the Wistar Institute (Philadelphia, USA) in Dr. Giorgio Trinchieri's and Bice Perussia’s laboratories, as Associate Scientist.

 

His studies had the purpose of clarifying:
- the nature, properties and the mechanisms of activation of the phagocyte NADPH oxidase.
- the influence of IFNgamma and other proinflammatory molecules on phagocyte oxidative metabolism.
- the mechanisms of activation of natural killer cells through CD16.
- gene expression during myeloid differentiation.
- gene expression during neutrophil activation.
- the effects of amiloyd beta (Abeta) on phagocyte proinflammatory functions.
- the molecular bases of the IL-10-mediated responses in human neutrophils.
- the regulatory mechanisms controlling neutrophil-mediated angiogenesis
- the molecular bases of cytokine production by human neutrophils.
- the signalling pathways activated via TLR4 in human leukocytes.
- the cross-talks that neutrophils undertake with other leukocytes

 


Current research interests

During the last years, Marco A. Cassatella's group has been involved in studies dealing with the mechanisms regulating, at molecular level, the effector functions of cells belonging to the innate system. In particular, his group has greatly contributed to uncover the notion that polymorphonuclear neutrophils (PMN) not only are a target of different cytokines (such as for instance IFNgamma or IL-15) able to modulate their effector functions, but also represent a source of a number of immunoregulatory molecules. In addition, Marco A. Cassatella's group has also concentrated his efforts in investigating the signal transduction pathways and the biochemical mechanisms involved in IL-10-mediated regulation of neutrophil gene expression. Current research interests are:

Molecular mechanisms regulating TLR4-mediated gene expression..
The innate-immune system is highly developed in its ability to discriminate between self and foreign pathogens, a process that relies, to a great extent, on an evolutionary conserved family of proteins, the Toll-like receptors (TLRs). Stimulation of the TLRs triggers complex signaling pathways, mainly characterized in the mouse system, containing branches that are common to all the TLRs and others that are selectively activated, which have the potential to tailor immune responses to pathogens, including proinflammatory cytokine production, dendritic cell (DC) maturation and polarized immune responses. Human neutrophils express mRNA for all the TLRs, except TLR3 and agonists of all the expressed TLRs elicit inflammatory responses in resting neutrophils, except for CpG, which requires a GM-CSF pretreatment. However, very little is known on the role and on the biochemical and molecular mechanisms mediated by TLRs that lead to the expression of cytokines or of other effector molecules in human neutrophils. In this regard, Cassatella's group is involved in a current research line, which stems from observations that the group published in 1997 (Eur J Immunol. 1997 Jan;27:111-5) and 1999 (J Immunol. 1999 Apr 15;162:4928-37). It was reported, in fact, that while monocytes are able to express CXCL10/IP-10 mRNA and to release the related protein in response either to LPS (a TLR4 ligand) alone, or, more efficiently, to IFNgamma, neutrophils upregulate CXCL10/IP-10 mRNA only if incubated with IFNgamma used together with LPS or TNFalfa. With the discovery of TLR4 as the putative receptor for LPS, a great effort has been made to characterize the molecular mechanisms whereby TLR4 triggers its biological effects, knockout mice being very helpful in this regard. From all these studies, which are continuously updated, it is well ascertained that, at least in mouse macrophages, LPS triggers two classes of genes via TLR4: one of the two (called “MyD88-dependent”) is rapidly and directly induced by endotoxin, and mostly includes proinflammatory mediators such as TNFalfa and CXCL8/IL-8; the other one (called MyD88-independent/IFNbeta-dependent, and including for instance CXCL10/IP-10) becomes expressed in a more delayed manner and is dependent on STAT1 activation, in turn activated by LPS-induced IFNbeta via the same pathway. Studies performed in the mouse system have clarified that distinct signal transduction pathways departing from TLR4 and controlled by specific adaptor proteins (namely MyD88, MAL/TIRAP, TRIF, TRAM), kinases (IRAK1-4, TBK1, IKKepsilon, etc) and transcription factors (NFkappaB, AP-1, IRF-3, etc) control/regulate these two classes of genes. On these preliminary bases, Cassatella's group is currently involved in clarifying the molecular mechanisms that in human neutrophils and monocytes regulate the expression of IFNbeta, CXCL10/IP-10 and other IFNbeta-dependent genes in response to TLR4 ligands. By using these experimental models, the group intends to also clarify whether the findings described in the mouse system, concerning the signalling cascades activated by TLR4 (and TLR3 as well), are similarly present and activated in human neutrophils and monocytes.

Molecular mechanisms controlling cytokine/gene expression in activated neutrophils.
The research conducted in recent years has brought forward exciting discoveries that have greatly broadened the knowledge on the functional role of neutrophils and uncovered novel links involving neutrophils to unsuspected physiopathologic processes. For instance, evidence on their active involvement in the resolution of inflammation (other than in its regulation), on their unquestionable regulatory role in angiogenesis and tumor fate, or else on their response to, and release of, a wide variety of cytokines and chemotactic molecules, have made it clear that the obsolete concept of the neutrophil as a "terminally differentiated, short-lived, cell devoid of transcriptional activities" found in most biomedical textbooks is certainly an old-style assumption which underestimates the multiple and amazing functional capacities of this cell type. At physiopathological level, neutrophils certainly have an important role not only as primary defensive effectors by exerting phagocytosis and bacteria destruction, but also as modulators of more complex processes, including the immune and antitumoral responses. During the last years, Cassatella's group has greatly contributed to clearly established that neutrophils are able to express, either in vitro or in vivo, not only several proinflammatory mediators, but also a number of immunoregulatory cytokines such as TNFalfa and IL-12, chemokines such as IL-8, GROalpha, IP-10, MIG, I-TAC, MIP-3alpha and MIP-3beta and cytokines with angiogenic and fibrogenic activities such as VEGF and HB-EGF (Adv Immunol. 1999; 73:369-509). To better comprehend the functional capacity of one of the most important component of the innate system and its potential attitude to interact with other arms of the immune system, it is of fundamental importance to characterize, at molecular level, the potential capacity of neutrophils to produce the various cytokine families, analyse in great detail the pattern of their expression and the regulatory mechanisms controlling their production, either in vitro or in various human inflammatory and neoplastic diseases.

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Characterization and pathological relevance of the reciprocal interactions between human neutrophils and immunoregulatory/effector cells of the innate and adaptive immune system.
It is now well established that neutrophils certainly have an important role not only as primary defensive effectors by exerting phagocytosis and bacteria destruction, but also as modulators of more complex processes, including the inflammatory, the immune, the antiinfectious and antitumoral response. These latter functions occur not only via neutrophil-derived cytokines, but also as a result of the potential capacity of neutrophils to interact with other cell types, which to date, represents an aspect of the neutrophil biology very poorly investigated and substantially unknown. Major objectives of the lab is to start exploring the effects that human neutrophils might potentially exert on the effector functions of NK cells, dendritic cells (DC), CD4+ and CD8+ T cells, as well as the recently identified Th17 cells. Preliminary results reveal the existence of significant reciprocal modulatory effects, of which we will try not only to identify the underlying molecular mechanisms, but also to confirm their biological meaning in selected infiammatory, autoimmune and infectious human pathologies. The identification of the networks underlying the cooperative role of neutrophils with the other leucocytes will provide targets for the prevention of pathological conditions without compromising their protective function.

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Molecular basis regulating IL-10 responsiveness in activated phagocytes.
IL-10 plays a key role in inhibiting neutrophil- and monocyte/macrophage-mediated inflammatory responses. However, the molecular mechanisms and the intracellular signalling pathways whereby IL-10 mediates its biological effects have not been clarified yet. In the last years, M.A. Cassatella has concentrated his efforts in investigating the biological effects of IL-10 on human neutrophils (particularly at the level of gene expression) and the signal transduction pathways involved in such activity. M.A.Cassatella is still actively involved in research dealing with the latter aspect. Recently, his group has demonstrated that, in myelomonocytic cells, the responsiveness to IL-10 is controlled by the regulated expression of IL-10R1 (J Immunol. 2001;167:2312-22), thus uncovering the existence of a negative feedback circuit in which, by upregulating IL-10R1 expression, either LPS, or other appropriate substances, render neutrophils and monocytic cells responsive to IL-10. In addition, his group has shown that Suppressor of Cytokine Signalling (SOCS)-3 is induced by IL-10 and, apparently inhibits LPS-induced phagocyte activation (Blood. 1999;94:2880-9; J Immunol. 2002;168:6404-11). More recently, Cassatella's group has provided evidence that LPS, other than upregulating the expression of the IL-10R1, necessitates to also induce (or constantly maintain) the synthesis of unknown component(s) involved in the IL-10-mediated signalling pathway (Eur J Immunol. 2005 May 2;35:1877-1885.). The latter one(s) appear to be necessary to facilitate the anti-inflammatory effects of IL-10 in LPS-stimulated neutrophils.

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LIST of SELECTED PUBLICATIONS

1. Cassatella MA, Anegon I, Cuturi MC, Griskey P, Trinchieri G, Perussia B. FcgammaR(CD16) interaction with ligand induces Ca2+ mobilization and phosphoinositide turnover in human NK cells. Role of Ca2+ in Fcgamma(CD16)-induced transcription and expression of lymphokine genes. J Exp Med. 1989;169(2):549-67.

2. Cassatella MA, Hartman L, Perussia B, Trinchieri G. TNF and IFNgamma synergistically induce cytochrome b-245 heavy-chain gene expression and nicotinamide-adenine dinucleotide phosphate hydrogenase oxidase in human leukemic myeloid cells. J Clin Invest. 1989, 83(5):1570-9.

3. Bazzoni F, Cassatella MA, Rossi F, Ceska M, Dewald B, Baggiolini M. Phagocytosing neutrophils produce and release high amounts of the NaP-1/interleukin 8. J Exp Med. 1991;173(3):771-4.

4. Baron P, Constantin G, D'Andrea A, Ponzin D, Scarpini E, Scarlato G, Trinchieri G, Rossi F, Cassatella MA. Production of TNF and other proinflammatory cytokines by human mononuclear phagocytes stimulated with myelin P2 protein. Proc Natl Acad Sci U S A. 1993;90(10):4414-8

5. Cassatella MA, Meda L, Bonora S, Ceska M, Constantin G. IL-10 inhibits the release of proinflammatory cytokines from human polymorphonuclear leukocytes. Evidence for an autocrine role of TNF and IL-1beta in mediating the production of IL-8 triggered by lipopolysaccharide. J Exp Med 1993;178(6):2207-11.

6. Cassatella MA, Meda L, Gasperini S, Calzetti F, Bonora S. IL-10 upregulates IL-1ra production from LPS-stimulated human PMN by delaying mRNA degradation. J Exp Med. 1994;179(5):1695-9.

7. Cassatella MA. The production of cytokines by neutrophils. Immunol Today. 1995;16(1):21-6

8. Meda L, Cassatella MA, Szendrei GI, Otvos L Jr, Baron P, Villalba M, Ferrari D, Rossi F. Activation of microglial cells by beta-amyloid protein and interferon-gamma. Nature. 1995;374(6523):647-50.

9. Cassatella MA. Neutrophil-derived proteins: selling cytokines by the pound. Adv Immunol. 1999;73:369-509.

10. Scapini P, Nardelli B, Nadali G, Calzetti F, Pizzolo G, Montecucco C, Cassatella MA. G-CSF-stimulated neutrophils are a prominent source of functional BLyS. J Exp Med. 2003;197(3):297-302.

11: Jaillon S, Peri G, Delneste Y, Frémaux I, Doni A, Moalli F, Garlanda C, Romani L, Bellocchio S, Bozza S, Cassatella MA, Jeannin P, Mantovani A. The humoral pattern recognition receptor PTX3 is stored in neutrophil granules and localizes in extracellular traps. J Exp Med. 2007;204:793-804.

12: Pelletier M, Maggi L, Micheletti A, Lazzeri E, Tamassia N, Costantini C, Cosmi L, Lunardi C, Annunziato F, Romagnani S, Cassatella MA. Evidence for a cross-talk between human neutrophils and Th17 cells. Blood 2010;115:335-43.

13: Cassatella MA, Locati M, Mantovani A. Never underestimate the power of a neutrophil. Immunity 2009;31:698-700.

14: Scapini P, Hu Y, Defranco AL, Cassatella MA, Lowell CA. Myeloid cells, BAFF, and IFNgamma establish an inflammatory loop that exacerbates autoimmunity in Lyn-deficient mice. J.Exp.Med 2010;207:1757-73.

15: Witko-Sarsat V, Mocek J, Bouayad D, Tamassia N, Ribeil JA, Candalh C, Davezac N, Reuter N, Mouthon L, Hermine O, Pederzoli-Ribeil M, Cassatella MA. Proliferating cell nuclear antigen acts as a cytoplasmic platform controlling human neutrophil survival. J Exp Med. 2010 Nov 22;207(12):2631-45.

16: Costantini C, Calzetti F, Perbellini O, Micheletti A, Scarponi C, Lonardi S, Pelletier M, Schakel K, Pizzolo G, Facchetti F, Vermi W, Albanesi C, Cassatella MA. Human neutrophils interact with both slanDC and NK cells to amplify NK-derived IFNg: role of CD18, ICAM-1, and ICAM-3. Blood 2011;117:1677-86.

17: Witko-Sarsat V, Pederzoli-Ribeil M, Hirsch E, Sozzani S, Cassatella MA. Regulating neutrophil apoptosis: new players enter the game. Trends Immunol. 2011 Mar;32(3):117-24.

18: Mantovani A, Cassatella MA, Costantini C, Jaillon S. Neutrophils in the activation and regulation of innate and adaptive immunity. Nat Rev Immunol. 2011;11:519-31.

 


 

Current Lab Personnel

Research Assistants
Patrizia Scapini

Postdoctoral Fellows
Nicola Tamassia - nicola_tamassia@hotmail.com
Deseada Camuesco - d.camuesco@qmul.ac.uk

Research Lab Assistants
Federica Calzetti - federica.calzetti@univr.it
Sara Gasperini - sara.gasperini@univr.it

Graduate Students
Alessandra Michieletti - alessandra.micheletti84@gmail.com
Maili Zimmermann - maili.zimmermann@ut.ee

 

person

From left (names with a "*" indicate lab members): Sara Gasperini, B.S., lab assistant, Marco A. Cassatella, Claudio Costantini, Ph.D., post doc*; Nicola Tamassia, Ph.D., post doc*; Flavia Bazzoni, Ph.D., Associate Professor; Federica Calzetti, B.S., lab assistant*; Marzia Rossato, post doc; Alessandra Micheletti, graduate student*.

 

Past Post Docs

Miguel Aste Amezaga (Peru’)

Flavia Bazzoni (Italy)

Corrada Bonaiuto (Italy)

Stefano Bonora (Italy)

Uta Bussmeyer (Germany)

Claudio Constantini (Italy)

Gabriela Constantin (Romania)

Luca Crepaldi (Italy)

Cristiane Furlaneto (Brasil)

Jose A. Lapinet Vera (Cuba)

Martina Marchi (Italy)

Martin Pelletier (Canada)

Vincent Le Moigne (France)

Patrick P. McDonald (Canada)

Lucia Meda (Italy)

Gabriela Pereira Da Silva (Brasil)

Diego Peroni (Italy)

Ana Persichini Rodriguez (Brasil)

Maria Pia Russo (Italy)

Cristina Tecchio (Italy)

Marieke Vollebreigt (The Netherlands)

 

Current and past Collaborators

C. Agostini (Padova, Italy), C. Albanesi (Roma, Italy), A. Albini (Genova, Italy), F. Annunziato (Firenze, Italy), R. Badolato (Brescia, Italy), M. Baggiolini (Bern, CH), LM Bambara (Verona, Italy), PL Baron (Milano, Italy), F. Bazzoni (Verona, Italy), S. Brandau (Essen, Germanyy), A. Campa (San Paolo, Brasil), M. Ceska (Vienna, Austria), M. Colonna (St.Louis, USA), M. De Bernard (Padua, Italy), R. De Waal Malefyt (Palo Alto, USA), S. Edwards (Liverpool, UK), F. Di Virgilio (Ferrara, Italy), M. Eberl (Cardiff, UK), J. Farber (Bethesda, USA), M. Krampera (Verona, Italy), M. Locati (Milano, Italy), M. Lopez-Botet (Barcelona, Spain), C. Lowell (San Francisco, USA), A. Luster (Boston, USA), A. Mantovani (Milano, Italy), PP. McDonald (Sherbrooke, Canada), S. Nedospasov (Frederick, USA), O. Perez (L’Avana, Cuba), G. Pizzolo (Verona, Verona), M. Presta (Brescia, Italy), L. Rivoltini (Milano, Italy), S. Romagnani (Firenze, Italy), K. Schakel (Heidelberg, Germany), G.P. Semenzato (Padova, Italy), S. Sozzani (Brescia, Italy), C. Tecchio (Verona, Italy), G. Trinchieri (Frederick, USA), W. Vermi (Brescia), V. Witko-Sarsat (Paris, France), A. Yoshimura (Fukuoka, Japan)