Which are the mechanisms regulating malignant B lymphocyte growth, fitness and resilience to cytotoxic agents?
Non-Hodgkin lymphomas (NHL) and multiple myeloma (MM) are B-cell derived tumours and represent the vast majority of hematologic cancers, in many instances still incurable. We aim at defining the determinants of intrinsic and extrinsic growth signals for NHL and MM cells. Our laboratory is particularly interested in identifying novel regulators of cellular fitness and stress resistance to be targeted in novel therapeutic approaches for NHL and MM.
What are the physiological roles of protein kinases CK1 and CK2 in hemolymphopoiesis?
Hematopoietic stem cell (HSC) self-renewal and differentiation in myeloid and lymphoid cells are processes tightly regulated by intrinsic and extrinsic signals. The Wnt/β-catenin, Hedgehog (Hh) and PI3K/AKT signaling pathways play a crucial role in HSC fate decision and biochemical and functional genetic studies have demonstrated their essential function in blood cell development. Protein kinases CK1α and CK2 are acidophilic serine-threonine kinases, which, despite structurally unrelated, share the property of impinging on the Wnt/β-catenin, Hh and PI3K/AKT signaling. CK1α regulates Wnt signalling by phosphorylating β-catenin on Ser45 and priming it for subsequent phosphorylation by GSK3β on Ser33, Ser37 and Thr41 in the β-catenin destruction complex. CK1α also regulates Hh signalling by phosphorylating Cubitus Interruptus/Gli proteins in concert with the kinases PKA and GSK3β, driving their proteolytic degradation and extinguishing Hh signalling. CK2 positively regulates Wnt signaling by phosphorylating Dishevelled and catenin at Thr393 and also regulates Hh signalling in a positive manner. In Drosophila, CK2 phosphorylates Smoothened, favoring the Hh-transduced signal and the downstream molecule Ci (the homolog of mammalian Gli), stabilizing it and overall causing a positive regulation. Moreover, the antagonistic actions of CK2 and B56-containing PP2A phosphatase might regulate the stability of Gli proteins through phosphorylating/dephosphorylating Daz interacting protein 1 (Dzip1), which has been shown pivotal for Gli proteins degradation (Figure 1). However, there are no studies that investigated the function of CK1α and CK2 in hematopoiesis. On these grounds, we have started a research program aimed at addressing the specific roles of CK1 and CK2 in normal hematopoietic cell development. We have set different experimental model systems including analysis of cell lines, normal hematopoietic cells from healthy human subjects and genetically modified mice. In particular, we have generated mice with LoxP sites flanking exon 1 of the CK1α gene that will be used to conditionally knockout CK1α in the hematopoietic and B-cell and T-cell compartments. We have also generated hematopoietic and B-cell specific knockout mice for the regulatory β subunit of CK2 using CK2β flox. These mice are currently under examination.
How are regulated the growth, stress resistance and fitness of NHL and MM cells?
MM is an incurable plasma cell malignancy, which causes a significant morbidity due to organ damage and bone tissue destruction. Non-Hodgkin lymphomas are a heterogeneous group of B-cell or T-cell derived blood malignancies that remain in large part incurable. Exploring the role of potential new regulators of these malignancies growth and survival, we were the first to demonstrate that protein kinases CK2 and GSK3 play an oncogenic function in MM. We showed that CK2 regulates the STAT3 and NF-B activation downstream from growth factors and cytokines whereas GSK3 modulates catenin and AKT levels in MM cells (Piazza F et al. Blood 2006, Piazza F et al. BMC Cancer 2010). Since we found that CK2 regulates the ER-stress/UPR in MM cells, we are currently investigating the mechanisms through which CK2 and the related kinases GSK3 and CK1 may control the NF-B, STAT3 and endoplasmic reticulum (ER) stress/unfolded protein response (UPR) signaling in MM and non-Hodgkin Lymphomas. We could prove that CK2 is a master regulator of a compensatory UPR response in malignant plasma cells by sustaining the activity of the kinase/endoribonuclease IRE1α and the function of the chaperoning complex composed by Heat shock protein 90 /Cell division cycle 37 (Hsp90/Cdc37) (Manni S et al. Clin Can Res 2012). Along this line, we are interested in checking whether mechanisms linking the DNA damage response, the ER stress response and the activation of signal transducers of a stress response could be regulated by the transcription factors XBP1, STAT3 and NF-B in NHL and MM In tight collaborations with the Division of Pathology of the University of Padova, we are also investigating the expression of these kinases, transcription factors and stress-regulating proteins in tissue samples from MM and NHL patients and the potential correlations with clinico-biological features of these diseases (Pizzi M, Piazza F et al. Oncotarget 2015). Moreover, we plan to set up a phospho-proteomic and proteomic program aimed at analysing the pattern in NHL and MM cells under basal and different stress conditions and manipulation of stress-related kinases.
What are the roles of CK1 and CK2 in B-cell receptor signalling?
Both “tonic” antigen (Ag)-independent, PI3K-dependent and “chronic active” NF-B-addicted B-cell receptor (BCR) signaling are crucial for the growth of most of non-Hodgkin lymphomas (NHL). The serine-threonine kinases CK1 and CK2 stimulate the PI3K/Akt/mTOR pathway and - downstream BCR - might interact with the Ikk-NF-B-activating Card11-Bcl10-MALT1 (CBM1) complex. We recently demonstrated high CK2 expression and activity in germinal center (GC)-derived NHL (Pizzi M, Piazza F et al. Oncotarget 2015) and preliminary data in knockout mice we have generated indicate that CK1 and CK2 are pivotal for early and late B cell development and may significantly impact on “tonic” as well as “active” BCR signalling (Zaffino F et al Blood 2014).
We are interested to: a) analyze the in vivo role of CK1 and CK2in normal and NHL B cells by using loss of function mouse and cellular models. b) disentangle the BCR signaling nodes where CK1 and CK2 are likely to be crucial: the PI3K/AKT/FOXOs and the CARD11/IKK/NF-B pathways and the CK2-regulated Hsp90/Cdc37 chaperoning function on signaling kinases. c) identify novel CK1 and CK2 target molecules in normal and NHL B lymphocytes by high throughput phosphoproteomics. d) test in mouse NHL cell xenograft models CK1 and CK2 inhibitors in conjunction with novel anti-lymphoma agents.
What is the role of CK1 and CK2 in autophagy and proteotoxic stress in NHL and MM cells?
Autophagy plays a fundamental role in B cell physiology and during malignant transformation. In MM, autophagy must be tightly regulated to keep cellular homeostasis (Pengo N et al. 2013; Oliva L and Cenci S, 2014). CK1 is downstream from RAS-induced autophagy maintaining the rate of the process under control, while CK2 is a crucial kinase in stimulating the activity of the phagosome cargo protein p62/SQSTM1. We are interested in assessing the pathophysiological roles of these two kinases in autophagy in NHL and MM. We have shown that CK1 regulates autophagy in MM since its inactivation triggers the autophagic flux (Carrino M et al, 2017). Protein kinases members of the CK1 family are under analysis in the autophagic pathways in NHL and MM. The regulation of autophagosome and correct cargoing activity by p62/SQSTM1 by CK2 is analysed in B cell tumors. The effects of the inactivation of CK1 and CK2 on the cytotoxicity on the protestasis exerted by proteasome inhibitors and immunomodulatory drugs (lenalidomide, thalidomide) and are investigated.
Laura Quotti Tubi
- In Chronic Lymphocytic Leukemia the JAK2/STAT3 Pathway Is Constitutively Activated and Its Inhibition Leads to CLL Cell Death Unaffected by the Protective Bone Marrow Microenvironment.
- Identification of the true hyperdiploid multiple myeloma subset by combining conventional karyotyping and FISH analysis. Barilà G, Bonaldi L, Grassi A, Martines A, Liço A, Macrì N, Nalio S, Pavan L, Berno T, Branca A, Calabretto G, Carrino M, Teramo A, Manni S, Piazza F, Semenzato G, Zambello R.Blood Cancer J. 2020 Feb 17;10(2):18. doi: 10.1038/s41408-020-0285-6.PMID: 32066724
- NK cells and CD38: Implication for (Immuno)Therapy in Plasma Cell Dyscrasias. Zambello R, Barilà G, Manni S, Piazza F, Semenzato G.Cells. 2020 Mar 21;9(3):768. doi: 10.3390/cells9030768.PMID: 32245149
- Actionable Strategies to Target Multiple Myeloma Plasma Cell Resistance/Resilience to Stress: Insights From "Omics" Research. Manni S, Fregnani A, Barilà G, Zambello R, Semenzato G, Piazza F.Front Oncol. 2020 May 15;10:802. doi: 10.3389/fonc.2020.00802. eCollection 2020.PMID: 32500036
- Clinical Characteristics and Outcome of West Nile Virus Infection in Patients with Lymphoid Neoplasms: An Italian Multicentre Study. Visentin A, Nasillo V, Marchetti M, Ferrarini I, Paolini R, Sancetta R, Rigolin GM, Cibien F, Riva M, Briani C, Marinello S, Piazza F, Gherlinzoni F, Krampera M, Bassan R, Cuneo A, Luppi M, Semenzato G, Marasca R, Trentin L. Hemasphere. 2020 Jun 8;4(3):e395. doi: 10.1097/HS9.0000000000000395. eCollection 2020 Jun.PMID: 32647801
- Primary pancreatic lymphoma: Clinical presentation, diagnosis, treatment, and outcome. Facchinelli D, Sina S, Boninsegna E, Borin A, Tisi MC, Piazza F, Scapinello G, Maiolo E, Hohaus S, Zamò A, Merli M, Stefani PM, Mellone F, Basso M, Sartori R, Rusconi C, Parisi A, Manfrin E, Krampera M, Ruggeri M, Visco C, Tecchio C.Eur J Haematol. 2020 Oct;105(4):468-475. doi: 10.1111/ejh.13468. Epub 2020 Jul 2.PMID: 32542880
- A case of "double hit" mantle cell lymphoma carrying CCND1 and MYC translocations relapsed/refractory to rituximab bendamustine cytarabine (R-BAC) and ibrutinib. Scapinello G, Riva M, Branca A, Pizzi M, Bonaldi L, Martines A, Manni S, Visentin A, Trentin L, Semenzato G, Gurrieri C, Piazza F.Ann Hematol. 2020 Nov;99(11):2715-2717. doi: 10.1007/s00277-020-04178-0. Epub 2020 Jul 15.PMID: 32671454
- Younger patients with Waldenström Macroglobulinemia exhibit low risk profile and excellent outcomes in the era of immunotherapy and targeted therapies. Varettoni M, Ferrari A, Frustaci AM, Ferretti VV, Rizzi R, Motta M, Piazza F, Merli M, Benevolo G, Visco C, Laurenti L, Ferrero S, Gentile M, Del Fabro V, Abbadessa A, Klersy C, Musto P, Fabbri N, Deodato M, Dogliotti I, Greco C, Corbingi A, Luminari S, Arcaini L.Am J Hematol. 2020 Dec;95(12):1473-1478. doi: 10.1002/ajh.25961. Epub 2020 Sep 9.PMID: 32780514 Clinical Trial.
- Ibrutinib in relapsed hairy cell leukemia variant: A case report and review of the literature. Visentin A, Imbergamo S, Trimarco V, Pravato S, Romano Gargarella L, Frezzato F, Scapinello G, Bertorelle R, Piva E, Facco M, Semenzato G, Piazza F, Trentin L.Hematol Oncol. 2020 Sep 26. doi: 10.1002/hon.2810. Online ahead of print.PMID: 32979282
- Lights and Shade of Next-Generation Pi3k Inhibitors in Chronic Lymphocytic Leukemia.Visentin A, Frezzato F, Severin F, Imbergamo S, Pravato S, Romano Gargarella L, Manni S, Pizzo S, Ruggieri E, Facco M, Brunati AM, Semenzato G, Piazza F, Trentin L.Onco Targets Ther. 2020 Sep 29;13:9679-9688. doi: 10.2147/OTT.S268899. eCollection 2020.PMID: 33061448
- Visentin A, Deodato M, Mauro FR, Autore F, Reda G, Vitale C, Molica S, Rigolin GM, Piazza F, Cesini L, Tedeschi A, Laurenti L, Cassin R, Coscia M, Cuneo A, Foà R, Semenzato G, Trentin L.Hematol Oncol. 2019 Oct;37(4):508-512. doi: 10.1002/hon.2655. Epub 2019 Aug 5.PMID: 31335982
- Merli M, Frigeni M, Alric L, Visco C, Besson C, Mannelli L, Di Rocco A, Ferrari A, Farina L, Pirisi M, Piazza F, Loustaud-Ratti V, Arcari A, Marino D, Sica A, Goldaniga M, Rusconi C, Gentile M, Cencini E, Benanti F, Rumi MG, Ferretti VV, Grossi P, Gotti M, Sciarra R, Tisi MC, Cano I, Zuccaro V, Passamonti F, Arcaini L. Direct-Acting Antivirals in Hepatitis C Virus-Associated Diffuse Large B-cell Lymphomas. Oncologist. 2019 Aug;24(8):e720-e729.
- Visco C, Tisi MC, Evangelista A, Di Rocco A, Zoellner AK, Zilioli VR, Hohaus S, Sciarra R, Re A, Tecchio C, Chiappella A, Morello L, Gini G, Nassi L, Perrone T, Molinari AL, Fabbri A, Cox MC, Finolezzi E, Ferrero S, Puccini B, Alvarez De Celis I, Arcari A, Marino D, Merli M, Piazza F, Gentile M, Pelosini M, Loseto G, Hermine O, Dreyling M, Ruggeri M, Martelli M, Hoster E, Vitolo U; Fondazione Italiana Linfomi and the Mantle Cell Lymphoma Network. Time to progression of mantle cell lymphoma after high-dose cytarabine-based regimens defines patients risk for death. Br J Haematol. 2019 Jun;185(5):940-944
- Carrino M, Quotti Tubi L, Fregnani A, Canovas Nunes S, Barilà G, Trentin L, Zambello R, Semenzato G, Manni S, Piazza F. Prosurvival autophagy is regulated by protein kinase CK1 alpha in multiple myeloma. Cell Death Discov. 2019 May 21;5:98
- Frezzato F, Raggi F, Martini V, Severin F, Trimarco V, Visentin A, Scomazzon E, Accordi B, Bresolin S, Piazza F, Facco M, Basso G, Semenzato G, Trentin L. HSP70/HSF1 axis, regulated via a PI3K/AKT pathway, is a druggable target in chronic lymphocytic leukemia. Int J Cancer. 2019 May 1.
- Visentin A, Imbergamo S, Scomazzon E, Pravato S, Frezzato F, Bonaldi L, Pizzi M, Vio S, Gregianin M, Burei M, Facco M, Semenzato G, Piazza F, Trentin L. BCR kinase inhibitors, idelalisib and ibrutinib, are active and effective in Richter syndrome. Br J Haematol. 2019 Apr;185(1):193-197.
- Barilà G, Compagno N, Liço A, Berno T, Bonaldi L, Teramo A, Manni S, Branca A, Frigo AC, Cinetto F, Piazza F, Semenzato G, Zambello R. Severe infections unrelated to neutropenia impact on overall survival in multiple myeloma patients: results of a single centre cohort study. Br J Haematol. 2019 Mar 10.
- Briani C, Visentin A, Campagnolo M, Salvalaggio A, Ferrari S, Cavallaro T, Manara R, Gasparotti R, Piazza F. Peripheral nervous system involvement in lymphomas J Peripher Nerv Syst. 2019 Mar;24(1):5-18.
- Pizzi M, Trentin L, Visentin A, Saraggi D, Martini V, Guzzardo V, Righi S, Frezzato F, Piazza F, Sabattini E, Semenzato G, Rugge M. Cortactin expression in non-Hodgkin B-cell lymphomas: a new marker for the differential diagnosis between chronic lymphocytic leukemia and mantle cell lymphoma. Hum Pathol. 2019 Mar;85:251-259
- Tisi MC, Paolini R, Piazza F, Ravelli E, Tecchio C, Sartori R, Famengo B, D'Amore ESG, Carli G, Perbellini O, Di Bona E, Ruggeri M, Visco C. Rituximab, bendamustine and cytarabine (R-BAC) in patients with relapsed-refractory aggressive B-cell lymphoma. Am J Hematol. 2018 Dec;93(12):E386-E389
- Visentin A, Campello E, Scomazzon E, Spiezia L, Imbergamo S, Pravato S, Piazza F, Semenzato G, Simioni P, Trentin L. Dabigatran in ibrutinib-treated patients with atrial fibrillation and lymphoproliferative diseases: Experience of 4 cases. Hematol Oncol. 2018 Dec;36(5):801-803.
- Visco C, Tisi MC, Evangelista A, Di Rocco A, Zoellner AK, Zilioli VR, Hohaus S, Sciarra R, Re A, Tecchio C, Chiappella A, Morello L, Gini G, Nassi L, Perrone T, Molinari AL, Fabbri A, Cox MC, Finolezzi E, Ferrero S, Puccini B, Alvarez De Celis I, Arcari A, Marino D, Merli M, Piazza F, Gentile M, Pelosini M, Loseto G, Hermine O, Dreyling M, Ruggeri M, Martelli M, Hoster E, Vitolo U; Fondazione Italiana Linfomi and the Mantle Cell Lymphoma Network. Time to progression of mantle cell lymphoma after high-dose cytarabine-based regimens defines patients risk for death. Br J Haematol. 2018 Nov 8.
- Kostic I, Ruiz M, Branca A, Nabergoj M, Piazza F, Semenzato G, Gurrieri C, Briani C. Possible neuroleukemiosis in two patients with acute myeloid leukemia in complete bone marrow remission. J Neurol Sci. 2018 Sep 15;392:63-64.
- Barilà G, Teramo A, Calabretto G, Ercolin C, Boscaro E, Trimarco V, Carraro S, Leoncin M, Vicenzetto C, Cabrelle A, Facco M, Piazza F, Semenzato G, Zambello R. Dominant cytotoxic NK cell subset within CLPD-NK patients identifies a more aggressive NK cell proliferation. Blood Cancer J. 2018 Jun 5;8(6):51
- Manni S, Carrino M, Semenzato G, Piazza F. Old and Young Actors Playing Novel Roles in the Drama of Multiple Myeloma Bone Marrow Microenvironment Dependent Drug Resistance. Int J Mol Sci. 2018 May 18;19(5). pii: E1512.
- Scapinello, G., Pizzi, M., Vio, S., Nabergoj, M., Visentin, A., Martines, A., Bonaldi, L., Trentin, L., Semenzato, G., Piazza, F. Splenic marginal zone lymphoma with a de novo t(8;14)(q24;q32) and a prolymphocytoid evolution responsive to rituximab-bendamustine (2018) Annals of Hematology, pp. 1-3.
- Canovas Nunes, S., Manzoni, M., Pizzi, M., Mandato, E., Carrino, M., Quotti Tubi, L., Zambello, R., Adami, F., Visentin, A., Barilà, G., Trentin, L., Manni, S., Neri, A., Semenzato, G., Piazza F. The small GTPase RhoU lays downstream of JAK/STAT signaling and mediates cell migration in multiple myeloma (2018) Blood Cancer Journal, 8 (2).
- Pizzi, M., Agostinelli, C., Righi, S., Gazzola, A., Mannu, C., Galuppini, F., Fassan, M., Visentin, A., Piazza F., Semenzato, G.C., Rugge, M., Sabattini, E. Aberrant expression of CD10 and BCL6 in mantle cell lymphoma (2017) Histopathology, 71 (5), pp. 769-777.
- Manni S, Carrino M, Piazza F. Role of protein kinases CK1α and CK2 in multiple myeloma: regulation of pivotal survival and stress-managing pathways. J Hematol Oncol. 2017 Oct 2;10(1):157. Review. PubMed PMID: 28969692; PubMed Central PMCID Oct 2017: PMC5625791.
- Mandato E, Nunes SC, Zaffino F, Casellato A, Macaccaro P, Tubi LQ, Visentin A,Trentin L, Semenzato G, Piazza F. CX-4945, a selective inhibitor of casein kinase 2, synergizes with B cell receptor signaling inhibitors in inducing diffuse large B cell lymphoma cell death. Curr Cancer Drug Targets. 2017 Apr 26. PubMed PMID: 28460620.
- Manni S, Carrino M, Manzoni M, Gianesin K, Nunes SC, Costacurta M, Tubi LQ, Macaccaro P, Taiana E, Cabrelle A, Barilà G, Martines A, Zambello R, Bonaldi L, Trentin L, Neri A, Semenzato G, Piazza F. Inactivation of CK1α in multiple myeloma empowers drug cytotoxicity by affecting AKT and β-catenin survival signaling pathways. Oncotarget. 2017 Feb 28;8(9):14604-14619. doi: 10.18632/oncotarget.14654. PubMed PMID: 28099937; PubMed Central PMCID: PMC5362429.
- Quotti Tubi L, Canovas Nunes S, Brancalion A, Doriguzzi Breatta E, Manni S, Mandato E, Zaffino F, Macaccaro P, Carrino M, Gianesin K, Trentin L, Binotto G, Zambello R, Semenzato G, Gurrieri C, Piazza F. Protein kinase CK2 regulates AKT, NF-κB and STAT3 activation, stem cell viability and proliferation in acute myeloid leukemia. Leukemia. 2017 Feb;31(2):292-300. doi: 10.1038/leu.2016.209. Epub 2016 Aug 1. PubMed PMID: 27479180.
- Mandato E, Manni S, Zaffino F, Semenzato G, Piazza F. Targeting CK2-driven non-oncogene addiction in B-cell tumors. Oncogene. 2016 Nov 24;35(47):6045-6052. doi: 10.1038/onc.2016.86. Epub 2016 Apr 4. Review. PubMed PMID: 27041560.
- Manni S, Toscani D, Mandato E, Brancalion A, Quotti Tubi L, Macaccaro P, Cabrelle A, Adami F, Zambello R, Gurrieri C, Semenzato G, Giuliani N, Piazza F. Bone marrow stromal cell-fueled multiple myeloma growth and osteoclastogenesis are sustained by protein kinase CK2. Leukemia. 2014 Oct;28(10):2094-7. doi: 10.1038/leu.2014.178. Epub 2014 Jun 4. PubMed PMID: 24897506.
- Manni S, Brancalion A, Mandato E, Quotti Tubi L, Colpo A, Pizzi M, Cappellesso R, Zaffino F, Di Maggio SA, Cabrelle A, Marino F, Zambello R, Trentin L, Adami F, Gurrieri C, Semenzato G, Piazza F (2013) Protein kinase CK2 inhibition down modulates the NF-kB and STAT3 survival pathways, enhances the cellular proteotoxic stress and synergistically boosts the cytotoxic effect of bortezomib on multiple myeloma and mantle cell lymphoma cells. PLOS One 2013 8: e75280.
- Manni S, Brancalion A, Quotti Tubi L, Colpo A, Pavan L, Cabrelle A, Ave E, Zaffino F, Di Maira G, Ruzzene M, Adami F, Zambello R, Pitari MR, Tasson P, Pinna LA, Gurrieri C, Semenzato G, Piazza F (2012) Protein kinase CK2 protects multiple myeloma cells from ER stress-induced apoptosis and from the cytotoxic effect of HSP90 inhibition through regulation of the unfolded protein response. Clin Cancer Res 2012 18:1888-900.
- Piazza F, Manni S, Ruzzene M, Pinna LA, Gurrieri C, Semenzato G (2012) Protein kinase CK2 in hematologic malignancies: reliance on a pivotal cell survival regulator by oncogenic signaling pathways. Leukemia 26:1174-1179.
- Piazza F, Manni, S, Quotti Tubi L, Montini B, Pavan L, Colpo A, Gnoato M, Cabrelle A, Adami F, Zambello R, Trentin L, Gurrieri C, Semenzato G (2010) Glycogen Synthase Kinase-3 regulates multiple myeloma cell growth and bortezomib-induced cell death. BMC Cancer 10:526.
- MD, University of Padova, Italy (1995)
- Board in Medical Oncology, University of Padova, Italy (2000)
- Postdoc: Memorial Sloan-Kettering Cancer Center, New York, USA (1999-2003)
- Assistant Professor of Hematology, Dept. of Medicine, University of Padua (since 2008)
- Group leader: Hematological Malignancies, Lymphoma and Myeloma Pathobiology, Venetian Institute of Molecular Medicine, Padua, Italy (since 2013)
- Associate Professor of Hematology, Dept. of Medicine, University of Padua (since 2018)
- 2004: Brian D. Novis Junior Award, International Myeloma Foundation (USA)
- 2006: “Guido Berlucchi” Award for young oncologists, Italy
- Gilead Fellowship