Massachusetts General Hospital

During the course of tumor progression, cancer cells acquire a number of characteristic alterations. These include the capacities to proliferate independently of exogenous growth promoting or growth-inhibitory signals, to invade surrounding tissues and metastasize to distant sites, to elicit an angiogenic response, and to evade mechanisms that limit cell proliferation, such as apoptosis and replicative senescence. These properties reflect alterations in the cellular signaling pathways that in normal cells control cell proliferation, motility, and survival. Many of the proteins currently under investigation as possible targets for cancer therapy are signaling proteins that are components of these pathways. Our laboratory is uses chemical biology approaches and models of epithelial tumorigenesis and regulation of keratinocytes proliferation to dissect such pathways.

We have recently identified a novel signaling pathway in keratinocytes involving inhibition of the Notch1 gene downstream of p53, which plays a key role in squamous cell carcinoma (SCC) development. Exploring the downstream effects of activated Notch receptor in the epidermis, we found that the small GTPase RhoE is a new transcriptional target of Notch1, which is essential for the differentiation switch in keratinocytes. RhoE deficiency in vitro and in vivo renders keratinocytes resistant to Notch1-mediated induction of differentiation thereby favoring uncontrolled growth and proliferation. Furthermore, we have strong evidence that RhoE binds to activated Notch1 and mediates the recruitment of the Notch1-transcriptional complex to the promoters of its target genes. Our working hypothesis is that RhoE is a key regulator of Notch1-mediated commitment to differentiation and suppression of carcinogenesis/tumorigenesis in the epidermis. The mechanistic understanding of the pathway(s) controlling the Notch-RhoE signaling cascade in the epidermis is expected to eventually translate into the development of therapeutics for the treatment of skin SCCs and other epithelial malignancies with down-modulated Notch signaling. It also brought to light an unexpected function of RhoE in the cellular response to Endoplasmic Reticulum Stress and thus paved the way for the discovery of novel modulators of these processes.

Our profound interest in Drug Development also complements the long-term goal of the laboratory to dissect specific stress-mediated responses of the cell and to develop unique ways to modify them. Mammalian Y-box binding protein-1 (YB-1) is a member of the DNA/RNA-binding family of proteins with an evolutionarily conserved cold-shock domain (CSD). Both bacterial and mammalian cold-shock domain proteins are ubiquitously expressed and involved in fundamental processes such as DNA repair, mRNA transcription, splicing, translation, and stabilization. Consistent with its essential biological functions, targeted disruption of YB-1 in mice causes severe developmental defects and embryonic lethality. Numerous studies point to a role for YB-1 in malignant transformation, with evidence for oncogenic functions. A pro-oncogenic role for YB-1 is suggested by its higher expression in actively proliferating tissues and multiple human malignancies, as well its ability to activate transcription of proliferation- related genes through binding to the Y-box promoter elements of the latter. We are currently studying the role of YB-1 in normal keratinocyte biology as well as in the pathogenesis of keratinocyte-derived cancers. In addition, we are also developing approaches for small molecule based inhibition of YB-1 expression and activity.

Lab Members:

Kristina Todorova, Ph.D.

Eunjeong Kwon, Ph.D.

Jun Wang

Anna Mandinova, M.D., Ph.D.

Publications:

1. Mandinova A, Atar D, Schäfer B, Spiess M, Aebi U and Heizmann CW.  Distinct Subcellular Localization of Calcium Binding S100 Proteins in Human Vascular Smooth Muscle Cells and Their Relocation in Response to Rises in Intracellular Calcium.  J Cell Sci 1998;  111 (Pt14):2043-54
2.  Atar D, Spiess M, Mandinova A, Cierpka H, Noll G, Lüscher TF. Carnitine--from cellular mechanisms to potential clinical applications in heart disease. Eur J Clin Invest. 1997 Dec;27(12):973-6.
3. Spiess M. Steinmetz MO, Mandinova A, Wolpensinger B, Aebi U, Atar D.  Isolation, electron microscopic imaging, and 3-D visulatization of native cardiac thin myofilaments.  J Struct Biol 1999;  126(2):98-104.
4. Schoenenberger CA, Steinmetz MO, Stoffler D, Mandinova A, Aebi U. Structure, assembly, and dynamics of actin filaments in situ and in vitro. Microsc Res Tech. 1999 Oct 1;47(1):38-50.
5. Baschong W, Duerrenberger M, Mandinova A, Suetterlin R. Three-dimensional visualization of cytoskeleton by confocal laser scanning microscopy. Methods Enzymol. 1999;307:173-89.
6. Brett W, Mandinova A, Remppis A, Sauder U, Rüter F, Heizmann CW, Aebi U, Zerkowski HR. Translocation of S100A1(1) calcium binding protein during heart surgery. Biochem Biophys Res Commun. 2001 Jun 15;284(3):698-703.
7. De La Cruz EM, Mandinova A, Steinmetz MO, Stoffler D, Aebi U, Pollard TD.  Polymerization and structure of nucleotide-free actin filaments.  J Mol Biol. 2000;  295(3):517-26.
8. Fahrenkrog B, Hubner W, Mandinova A, Pante N. Keller W, Aebi U.  The yeast nucleoporin Nup53p specifically interacts with Nic96p and is directly involved in nuclear protein import.  Mol Biol Cell 2000;  11(11):3885-96.
9. Klainguti M, Aigner S. Kilo J, Eppenberger HM, Mandinova A, Aebi U, Schaub MC, Shaw SG, Luscher TF, Atar D.  Lack of nuclear apoptosis in cardiomyocytes and increased endothelin-1 levels in a rat heart model of myocardial stunning.  Basic Res Cardiol. 2000;  95(4):308-15.
10. Maco B, Mandinova A, Durrenberger MB, Schafter BW, Uhrik B, Heizmann CW.  Ultrastructual distribution of the S100A1 Ca2+- binding protein in the human heart.  Physiol Res.  2001;  50(6):567-74.
11. Landriscina M, Bagala C, Mandinova A, Soldi R, Micucci I, Bellum S, Prudovsky I, Maciag T.  Copper induces the assembly of a multiprotein aggregate implicated in the release of fibroblast growth factor 1 in response to stress.  J Biol Chem. 2001;  276(27):25549-57.
12. Yang Z, Mandinova A, Kozai T, Joch H, Aebi U, Luscher TF.  Felodipine inhibits nuclear translocation of p42/44 mitogen-activated protein kinase and human smooth muscle cell growth. Cardiovasc Res. 2002 Jan;53(1):227-31.
13. Prudovsky I, Bagala C, Tarantini F, Mandinova A, Soldi R, Bellum S, Maciag T.  The intracellular translocation of the components of the fibroblast growth factor 1 release complex precedes their assembly prior to export.  J. Cell Biol 2002;  158(2):201-8.
14. Small D, Kovalenko D, Soldi R, Mandinova A, Kolev V, Trifonaova R, Bagala C, Kacer D, Battelli C, Liaw L, Prudovsky I, Maciag T.  Notch activation suppresses fibroblast growth factor-dependent cellular tranformation.  J Biol Chem 2003;  278(18):16405-13.
15. Mandinova A, Soldi R, Graziani I, Bagala C, Bellum S, Landriscina M, Taratini F, Prudovsky I, Maciag T.  S100A13 mediates the copper-dependent stress-induced release of IL-1{alpha} from both human U937 and murine NIH3T3 cells.  J Cell Sci 2003;  116(pt 13):2687-2696.
16. Mandinov L*, Mandinova A.*, Kyurkchiev S, Kyurkchiev D, Kehayov I, Kolev V, Soldi R, Bagala C, De Muinck ED, Lindner V, Post MJ, Simons M, Bellum S, Prudovsky I, Maciag T.  Copper chelation represses the vascular response to injury.  Proc Natl Acad Sci USA 2003;  100(11):6700-5.
17. Bagala C, Kolev V, Mandinova A, Soldi R, Mouta C, Graziani I, Prudowsky I, Maciag T.  The alternative translation of synaptotagmin 1 mediates the non-classical release of FGF1.  Biochem Biophys Res Commun 2003;  310(4):1041-7.
18. Mandinov L, Mandinova A, Soldi R, Graziani I, Bagala C, Prudovsky I, Maciag T. Interleukin 1: the choreographer for the restenotic ballet. Thromb Haemost. 2003 Sep;90(3):369-71.
19. Trifonova R, Small D, Kacer D, Kovalenko D, Kolev V, Mandinova A, Soldi R, Liaw L, Prudovsky I, Maciag T.  The non-transmembrane form of Delta1 but not of jagged1 Induces normal migratory behavior accompanied by FGF receptor 1-dependent transformation.  J Biol Chem. 2004 Apr 2;279(14):13285-8
20. Mourdjeva, M, Kyurkchiev D, Mandinova A, Altankova I, Kehayov I, Kyurkchiev S. Dynamics of membrane translocation of phosphatidylserine during apoptosis detected by a monoclonal antibody. Apoptosis. 2005 Jan;10 (1):209-17.
21. Nguyen B, Lefort K, Mandinova A, Devgan V, Antonini D, Koster M, Wang J, Zhang Z, Tommasi di Vignano A, Kitajewski J, Chiorino G, Roop D, Missero C, and Dotto GP. Cross-regulation between Notch and p63 in keratinocyte commitment towards differentiation. Genes Dev. 2006 Apr 15;20 (8):1028-42.
22. Ayyanan A, Civenni G, Ciarloni L, Morel C, Mueller N, Lefort K, Mandinova A, Raffoul W, Fiche M, Dotto GP, Brisken C. Increased Wnt signaling triggers oncogenic conversion of human breast epithelial cells by a Notch-dependent mechanism. Proc Natl Acad Sci U S A. 2006 Mar 7; 103(10):3799-804.
23. Mandinov L, Moodie KL, Mandinova A, Zhuang Z, Redican F, Baklanov D, Lindner V, Maciag T, Simons M, de Muinck ED. Inhibition of In-Stent Restenosis by Oral Copper Chelation in Porcine Coronary Arteries. Am J Physiol Heart Circ Physiol. 2006 Dec;291(6):H2692-7
24. Lefort K, Mandinova A, Ostano P, Kolev V, Calpini V, Kolfschoten I, Devgan V, Lieb J, Raffoul W, Hohl D, Neel V, Garlick J, Chiorino G, Dotto GP. Notch1 is a p53 target gene involved in human keratinocyte tumor suppression through negative regulation of ROCK1/2 and MRCKalpha kinases; Genes Dev. 2007 Mar 1;21(5):562-77
25.  Soldi R, Mandinova A, Venkataraman K, Hla T, Vadas M, Pitson S, Duarte M, Graziani I, Kolev V, Kacer D, Kirov A, Maciag T, Prudovsky I.            Sphingosine kinase 1 is a critical component of the copper-dependent FGF1 export pathway. Exp Cell Res. 2007 Sep 10;313(15):3308-18
26.  Brown L, Ongusaha PP, Kim HG, Nuti S, Mandinova A, Lee JW, Khosravi-Far R, Aaronson SA, Lee SW. CDIP, a novel pro apoptotic gene, regulates TNFalpha-mediated apoptosis in a p53-dependent manner. EMBO J. 2007 Jul 25;26(14):3410-22.
27. Antonini D, Dentice M, Mahtani P, De Rosa L, Della Gatta G, Mandinova A, Salvatore D, Stupka E, Missero C Tprg, a gene predominantly expressed in skin, is a direct target of the transcription factor p63J Invest Dermatol. 2008 Jul;128(7):1676-85.
28. Mandinova A,*, Karine Lefort*, Alice Tommasi di Vignano, Wesley Stonely, Paola Ostano, Giovanna Chiorino, Haruhi Iwaki, Jotaro Nakanishi and G. Paolo Dotto; FoxO3a is a key transcriptional target of canonical Notch signaling in the keratinocyte UVB-response. EMBO J. 2008 Apr 23;27(8):1243-54
29. Kolev V*, Mandinova A*, Guinea-Viniegra J, Hu B, Lefort K, Lambertini C, Neel V, Dummer R, Wagner EF, Dotto GP.EGFR signalling as a negative regulator of Notch1 gene transcription and function in proliferating keratinocytes and cancer. Nat Cell Biol. 2008 Aug;10(8):902-11.
30. Stanton BZ, Peng LF, Maloof N, Nakai K, Wang X, Duffner JL, Taveras KM, Hyman JM, Lee SW, Koehler AN, Chen JK, Fox JL, Mandinova A, Schreiber SL. A small molecule that binds Hedgehog and blocks its signaling in human cells. Nat Chem Biol. 2009 Mar;5(3):154-6. Epub 2009 Jan 18
31. Mandinova A Kolev V, Neel V, Hu B, Stonely W, Lieb J, Wu X, Colli C, Han R, Pazin M, Ostano P, Dummer R, Brissette JL, Dotto GP. A positive FGFR3/FOXN1 feedback loop underlies benign skin keratosis versus squamous cell carcinoma formation in humans. J Clin Invest. 2009 Oct;119(10):3127-37.
32. Lee KK, Todorova K, Mandinova A. Maximizing early detection of esophageal squamous cell carcinoma via SILAC-proteomics. Cancer Biol Ther. 2010 Oct 15;10(8):811-3. Epub 2010 Oct 15.
33. Mandinova A, Lee SW. The p53 pathway as a target in cancer therapeutics: obstacles and promise. Sci Transl Med. 2011 Jan 5;3(64):64rv1.
34. Kim HG, Hwang SY, Aaronson SA, Mandinova A, Lee SW. DDR1 receptor tyrosine kinase promotes prosurvival pathway through Notch1 activation. J Biol Chem. 2011 May 20;286(20):17672-81. Epub 2011 Mar 13.
35. Lakshmi Raj,¹ Takao Ide,¹ Aditi U Gurkar,¹ Monica Schenone,² Guo Wei,² Nicola J. Tolliday,² Andrew M. Stern,² Todd R. Golub,² Steven A. Carr,² Alykhan F. Shamji,² Michael Foley,² Anna Mandinova,^1,2* Stuart L. Schreiber,^2* and Sam W. Lee^1,2*  Selective killing of cancer cells by enhancing oxidative stress response; Nature. 2011 Jul 13;475(7355):231-4.
36. Kawasumi M, Lemos B, Bradner JE, Thibodeau R, Kim YS, Schmidt M, Higgins E, Koo SW, Angle-Zahn A, Chen A, Levine D, Nguyen L, Heffernan TP, Longo I, Mandinova A, Lu YP, Conney AH, Nghiem P. Protection from UV-induced skin carcinogenesis by genetic inhibition of the ataxia telangiectasia and Rad3-related (ATR) kinase. Proc Natl Acad Sci U S A. 2011 Aug 16;108(33):13716-21. Epub 2011 Aug 15.

Cutaneous Biology Research Center

Phone: 617-726-4354