Scientific interests

The adult testis contains a small pool of stem cells called spermatogonial stem cells which have been isolated from both human and murine testis. Cultured spermatogonial stem cells derived from adult murine testis can undergo a spontaneous conversion process at low efficiency into pluripotent stem cells. These germline cell-derived pluripotent stem cells or GPSCs are similar in appearance to mouse ES cells. GPSCs can be continuously passaged under appropriate culture conditions without decline in colony forming ability or change in karyotype. These cells can also be cryopreserved and thawed with no loss in proliferation or differentiation capacity. And most importantly, no peculiar manipulation is required for “reprogramming” these adult stem cells. The GPSCs have been induced to differentiate into functional cardiomyocytes, neurons and glia in vitro. GPSCs are of prime importance in tissue regeneration due to the common features with both ES cells and adult stem cells. The possibility that GPSCs may contribute to liver or kidney holds great promise for the treatment of diseases affecting these organs.
The unit has already developed a system for differentiating GPSCs into functional hepatocytes with high efficiency through embryoid body formation (Fagoonee et al., Stem Cells and Develolment 2010). These hepatocytes were characterised at cellular, molecular and functional levels. The GPSC-derived hepatocytes expressed hepatic markers and were metabolically active as shown by albumin and haptoglobin secretion, urea synthesis, glycogen storage and indocyanine green uptake. We also performed an unprecedented DNA microarray analysis comparing different stages of hepatocyte differentiation. Gene expression profiling demonstrated a strong similarity between GPSC and ES cells at different stages of induced hepatic differentiation. Moreover, Pearson correlation analysis of the microarray datasets suggested that, at late hepatic differentiation stages, the in vitro-derived cells were closer to fetal mouse primary hepatocytes than to those obtained from neonates. Future studies will show whether GPSC-derived hepatocytes have the ability to restore hepatic function in disease models, and hence offer the opportunity to produce immune-matched hepatocytes for patient-specific treatment.

Main aims

1. Develop conditions for the isolation and culture of murine and human spermatogonial stem cells for the derivation of germline-derived pluripotent stem cells (GPSCs).

2. Differentiate GPSCs into functional cell types, other than hepatocytes, in vitro.

3. Use the differentiated and functional cells obtained from GPSCs in cell-based therapies in animal models of hemochromatosis and glycogenosis and then extend the research to human. We have competence in mouse surgery (hepatectomy, renal ischemia and reperfusion injury, as well as other drug-induced acute liver and kidney injuries).

3. Employ a bioinformatics approach to identify genes that are enhancers of reprogramming. Research in this field is currently under way.

Figure showing a colony of germline-derived pluripotent stem cells (GPSCs) in culture (A) and hepatocyte-like cells that have been induced to differentiate from these GPSCs (B).

Stem Cells

1. Generation of Functional Hepatocytes from Mouse Germline Cell-derived Pluripotent Stem Cells in vitro. Fagoonee S, Hobbs RM, De Chiara L, Cantarella D, Piro RM, Tolosano E, Medico E, Provero P, Pandolfi PP, Silengo L, Altruda F. Stem Cells Dev. 2010 Mar 23. [Epub ahead of print]

2. A dive into the stem cells world. Fagoonee S, Pellicano R, Altruda F. Panminerva Med. 2009 Mar;51(1):1-3.

3. Tissue stem cells and cancer stem cells: potential implications for gastric cancer.Gumucio DL, Fagoonee S, Qiao XT, Liebert M, Merchant JL, Altruda F, Rizzetto M, Pellicano R. Panminerva Med. 2008 Mar;50(1):65-71. Review.

Iron Metabolism

4. Heme-hemopexin complex attenuates neuronal cell death and stroke damage.Li RC, Saleem S, Zhen G, Cao W, Zhuang H, Lee J, Smith A, Altruda F, Tolosano E, Doré S. J Cereb Blood Flow Metab. 2009 May;29(5):953-64. Epub 2009 Mar 11.

5. Comparison of 3 Tfr2-deficient murine models suggests distinct functions for Tfr2-alpha and Tfr2-beta isoforms in different tissues.Roetto A, Di Cunto F, Pellegrino RM, Hirsch E, Azzolino O, Bondi A, Defilippi I, Carturan S, Miniscalco B, Riondato F, Cilloni D, Silengo L, Altruda F, Camaschella C, Saglio G. Blood. 2010 Apr 22;115(16):3382-9. Epub 2010 Feb 23.

6. Hemopexin affects iron distribution and ferritin expression in mouse brain.Morello N, Tonoli E, Logrand F, Fiorito V, Fagoonee S, Turco E, Silengo L, Vercelli A, Altruda F, Tolosano E. J Cell Mol Med. 2008 Dec 16.

7. Lack of plasma protein hemopexin dampens mercury-induced autoimmune response in mice. Fagoonee S, Caorsi C, Giovarelli M, Stoltenberg M, Silengo L, Altruda F, Camussi G, Tolosano E, Bussolati B. J Immunol. 2008 Aug 1;181(3):1937-47.

8. Hemopexin prevents endothelial damage and liver congestion in a mouse model of heme overload. Vinchi F, Gastaldi S, Silengo L, Altruda F, Tolosano E. Am J Pathol. 2008 Jul;173(1):289-99. Epub 2008 Jun 13.

9. Lack of haptoglobin affects iron transport across duodenum by modulating ferroportin expression. Marro S, Barisani D, Chiabrando D, Fagoonee S, Muckenthaler MU, Stolte J, Meneveri R, Haile D, Silengo L, Altruda F, Tolosano E. Gastroenterology. 2007 Oct;133(4):1261-1271.

10. Microarray and large-scale in silico--based identification of genes functionally related to Haptoglobin and/or Hemopexin. Fagoonee S, Di Cunto F, Vozzi D, Volinia S, Pellegrino M, Gasparini P, Silengo L, Altruda F, Tolosano E. DNA Cell Biol. 2006 Jun;25(6):323-30.

11. Hemoglobin and heme scavenging. Ascenzi P, Bocedi A, Visca P, Altruda F, Tolosano E, Beringhelli T, Fasano M. IUBMB Life. 2005 Nov;57(11):749-59. Review.

12. Plasma protein haptoglobin modulates renal iron loading. Fagoonee S, Gburek J, Hirsch E, Marro S, Moestrup SK, Laurberg JM, Christensen EI, Silengo L, Altruda F, Tolosano E. Am J Pathol. 2005 Apr;166(4):973-83.

13. Haptoglobin modifies the hemochromatosis phenotype in mice. Tolosano E, Fagoonee S, Garuti C, Valli L, Andrews NC, Altruda F, Pietrangelo A. Blood. 2005 Apr 15;105(8):3353-5. Epub 2004 Dec 21.

14. Enhanced splenomegaly and severe liver inflammation in haptoglobin/hemopexin double-null mice after acute hemolysis. Tolosano E, Fagoonee S, Hirsch E, Berger FG, Baumann H, Silengo L, Altruda F. Blood. 2002 Dec 1;100(12):4201-8. Epub 2002 Jul 25.

15. Hemopexin: structure, function, and regulation. Tolosano E, Altruda F. DNA Cell Biol. 2002 Apr;21(4):297-306. Review. Defective recovery and severe renal damage after acute hemolysis in hemopexin-deficient mice. Tolosano E, Hirsch E, Patrucco E, Camaschella C, Navone R, Silengo L, Altruda F. Blood. 1999 Dec 1;94(11):3906-14.

Miscellaneous

16. Therapeutic efficacy and immunological response of CCL5 antagonists in models of contact skin reaction. Canavese M, Altruda F, Silengo L. PLoS One. 2010 Jan 15;5(1):e8725.

17. Essential role of the p110beta subunit of phosphoinositide 3-OH kinase in male fertility. Ciraolo E, Morello F, Hobbs RM, Wolf F, Marone R, Iezzi M, Lu X, Mengozzi G, Altruda F, Sorba G, Guan K, Pandolfi PP, Wymann MP, Hirsch E. Mol Biol Cell. 2010 Mar;21(5):704-11. Epub 2010 Jan 6. Leukocyte transmigration is modulated by chemokine-mediated PI3Kgamma-dependent phosphorylation of vimentin. Barberis L, Pasquali C, Bertschy-Meier D, Cuccurullo A, Costa C, Ambrogio C, Vilbois F, Chiarle R, Wymann M, Altruda F, Rommel C, Hirsch E. Eur J Immunol. 2009 Apr;39(4):1136-46.

18. Phosphoinositide 3-kinase p110beta activity: key role in metabolism and mammary gland cancer but not development. Ciraolo E, Iezzi M, Marone R, Marengo S, Curcio C, Costa C, Azzolino O, Gonella C, Rubinetto C, Wu H, Dastrù W, Martin EL, Silengo L, Altruda F, Turco E, Lanzetti L, Musiani P, Rückle T, Rommel C, Backer JM, Forni G, Wymann MP, Hirsch E. Sci Signal. 2008 Sep 9;1(36):ra3.

19. The RhoA-associated protein Citron-N controls dendritic spine maintenance by interacting with spine-associated Golgi compartments. Camera P, Schubert V, Pellegrino M, Berto G, Vercelli A, Muzzi P, Hirsch E, Altruda F, Dotti CG, Di Cunto F. EMBO Rep. 2008 Apr;9(4):384-92. Epub 2008 Feb 29.

20. Structure of msj-1 gene in mice and humans: a possible role in the regulation of male reproduction. Meccariello R, Berruti G, Chianese R, De Santis R, Di Cunto F, Scarpa D, Cobellis G, Zucchetti I, Pierantoni R, Altruda F, Fasano S. Gen Comp Endocrinol. 2008 Mar 1;156(1):91-103. Epub 2007 Dec 4.

21. Negative feedback regulation of Rac in leukocytes from mice expressing a constitutively active phosphatidylinositol 3-kinase gamma. Costa C, Barberis L, Ambrogio C, Manazza AD, Patrucco E, Azzolino O, Neilsen PO, Ciraolo E, Altruda F, Prestwich GD, Chiarle R, Wymann M, Ridley A, Hirsch E. Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14354-9. Epub 2007 Aug 24.

22. p130Cas as a new regulator of mammary epithelial cell proliferation, survival, and HER2-neu oncogene-dependent breast tumorigenesis. Cabodi S, Tinnirello A, Di Stefano P, Bisarò B, Ambrosino E, Castellano I, Sapino A, Arisio R, Cavallo F, Forni G, Glukhova M, Silengo L, Altruda F, Turco E, Tarone G, Defilippi P. Cancer Res. 2006 May 1;66(9):4672-80.

23. Cardiac overexpression of melusin protects from dilated cardiomyopathy due to long-standing pressure overload. De Acetis M, Notte A, Accornero F, Selvetella G, Brancaccio M, Vecchione C, Sbroggiò M, Collino F, Pacchioni B, Lanfranchi G, Aretini A, Ferretti R, Maffei A, Altruda F, Silengo L, Tarone G, Lembo G. Circ Res. 2005 May 27;96(10):1087-94. Epub 2005 Apr 28. Erratum in: Circ Res. 2005 Jul 8;97(1):e5.

24. Protection from angiotensin II-mediated vasculotoxic and hypertensive response in mice lacking PI3Kgamma. Vecchione C, Patrucco E, Marino G, Barberis L, Poulet R, Aretini A, Maffei A, Gentile MT, Storto M, Azzolino O, Brancaccio M, Colussi GL, Bettarini U, Altruda F, Silengo L, Tarone G, Wymann MP, Hirsch E, Lembo G. J Exp Med. 2005 Apr 18;201(8):1217-28. Epub 2005 Apr 11.

25. PI3Kgamma modulates the cardiac response to chronic pressure overload by distinct kinase-dependent and -independent effects. Patrucco E, Notte A, Barberis L, Selvetella G, Maffei A, Brancaccio M, Marengo S, Russo G, Azzolino O, Rybalkin SD, Silengo L, Altruda F, Wetzker R, Wymann MP, Lembo G, Hirsch E. Cell. 2004 Aug 6;118(3):375-87.

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