**Simone Difilippantonio (formerly Petersen), Ph.D.**

**Mailing address: Contact Information:**

13902 Flint Rock Rd. Phone: (301) 460-0404

Rockville, MD 20853 e-mail: difilips@mail.nih.gov

**[EDUCATION:]{.underline}**

> 1996 -- Diploma (Biochemistry), Humboldt University, Berlin, Germany

1999 -- Ph.D. (Genetics), Humboldt University, Berlin, Germany

**[PROFESSIONAL AND RESEARCH EXPERIENCE:]{.underline}**

> May 1995 - April 1996: Diploma student, Institute of Pathology,
> Charite University Hospital, Berlin, Germany (PD Dr. Thomas von
> Zglinicki)
>
> May 1996 -- August 1999: PhD student, Institute of Pathology, Charite
> University Hospital, Berlin, Germany (PD Dr. Iver Petersen)
>
> September 1999 - August 2000: Research Scientist, Institute of
> Pathology, Charite University Hospital, Berlin, Germany (Prof. Dr.
> Manfred Dietel)
>
> September 2000- July 2003: Postdoctoral fellow, Experimental
> Immunology Branch, National Cancer Institute, National Institutes of
> Health, Bethesda, MD (Dr. Andre Nussenzweig)
>
> August 2003- February 2006: Research fellow, Experimental Immunology
> Branch, National Cancer Institute, National Institutes of Health,
> Bethesda, MD (Dr. Andre Nussenzweig)
>
> March 2006- Present: Staff Scientist, Experimental Immunology Branch,
> National Cancer Institute, National Institutes of Health, Bethesda, MD
> (Dr. Andre Nussenzweig)

**1995-1996:**  [Diploma Student, Institute of Pathology, Charite
University Hospital, Advisor: Dr. Thomas von Zglinicki.]{.underline}

*Telomere shortening*

                     I studied the effect of DNA damaging agents such as
oxidative stress on telomere shortening (see publication).

**1996-2000:** [Doctoral Student and Research Scientist, Institute of
Pathology, Charite University Hospital, Dr. Iver Petersen/Prof. Manfred
Dietel.]{.underline}

*Characterization of chromosomal changes during lung-cancer progression*

I studied genetic aberrations in lung and head and neck carcinomas using
multiplex-FISH, comparative genomic hybridization, and gene expression
profiling. Results from these studies have been published in numerous
papers and indicated that certain chromosomal aberrations are correlated
with disease stage and progression making them useful diagnostic
markers. To further identify the actual genes affected by these
aberrations, I cloned hundreds of differentially expressed genes that
have been shown to be involved in carcinogenesis (see publications).

In addition to research, I was the laboratory manager and involved in
developing scientific collaborations, applying for research grants,
laboratory setup, hiring/training and supervision of technicians and
doctoral students.

**2000-2006**: [Postdoctoral and Research Fellow, National Cancer
Institute. Advisor: Dr. Andre Nussenzweig.]{.underline}

> *DNA damage response, repair and maintenance of genomic stability.*
>
> My research centered on elucidating how normal cells maintain genomic
> stability. In particular, I studied the interplay between DNA damage
> detection, signaling and repair proteins and the role of DNA damage
> proteins in the maintenance of genomic stability and prevention of
> oncogenic translocations. Using several mouse models that colleagues
> and I generated, we were able to show that the DNA damage response is
> activated by antigen receptor rearrangement during B and T cell
> development. We showed that defects in proteins involved in the DNA
> damage response such as ATM, NBS1, H2AX, 53BP1 are required for
> efficient lymphocyte development and prevention of lymphomagenesis.
> One focus of my work was to study the role of Nbs1 in vivo through the
> development of a conditional knockout mouse. By restricting Nbs1
> deletion to B lymphocytes, I discovered that loss of the encoded
> protein is not only a downstream target of AT mutated (ATM) kinase but
> also acts upstream, promoting optimal ATM activation, recruitment to
> breaks, and accessibility to substrates. These results explain the
> clinical overlap of patients with Ataxia-Telengiectasia and Nijmegen
> Breakage Syndrome. By reconstituting Nbs1 knockout mice with bacterial
> artificial chromosomes (BACs) containing mutations in functional
> domains of the human Nbs1 gene, we generated the first humanized mouse
> models recapitulating the patient phenotype characterized by
> lymphocyte and germ cell developmental defects, inefficient cell-cycle
> arrest upon DNA damage induction, genomic instability and cancer
> predisposition (see publications).

**2006-present**: [Staff Scientist, National Cancer Institute. Advisor:
Dr. Andre Nussenzweig.]{.underline}

> *DNA damage response, repair and maintenance of genomic stability.*
>
> I continue to generate new and utilize existing mouse models to study
> the role of the aforementioned proteins during development and
> tumorigenesis. My position carries the additional responsibility
> contributing intellectually to the direction of the lab as well as the
> training and supervision of technicians and post-docs.

[TECHNICAL EXPERIENCE:]{.underline}

-   DNA extraction, Southern blot analysis, PCR, Quantitative-PCR

-   RNA-Extraction, Northern blot analysis, RT-PCR, Quantitative-RT-PCR,
    cloning of cDNA libraries

-   Establishment of cell lines from primary tumors (mouse, human),
    culture of primary and immortalized cells, generation of mouse
    embryonic fibroblasts (MEFs), Embryonic Stem cell cultures

-   Isolation, culture and analysis of murine T and B cells from lymph
    node, spleen, bone marrow

-   Transient and stable transfections, Microcell mediated chromosome
    transfer (MMCT), growth curves, survival, cytotoxicity assays,
    soft-agar growth, nude mice assays

-   Molecular cytogenetic techniques: Fluorescence in situ hybridization
    (FISH), Comparative Genomic Hybridization (CGH), Spectral
    Karyotyping (SKY), 3D-FISH

-   Immunocytochemistry, Immunofluorescence (IF), combined IF/FISH

-   Microscopy: Brightfield, Fluorescence, confocal imaging, live-cell
    imaging

-   Mutational analysis: microsatellite and SSCP analysis, sequencing

-   Design and cloning of targeting vectors for generation of transgenic
    and knockout/conditional knockout mice, conventional cloning,
    Bacterial Artificial Chromosome (BAC) recombineering, site-directed
    mutagenesis

-   Protein characterization: protein extraction, Western blot analysis,
    chromatin fractionation, ELISA, ELISPOT

-   Fluorescence activated cell sorting (FACS), cell-cycle analysis,
    apoptosis assays

**[AWARDS:]{.underline}**

> Fellowship Advancement Award 2002
>
> Fellows Award for Research Excellence 2003

**[BIBLIOGRAPHY]{.underline}**

***Peer reviewed articles:***

1.  Bockmuhl, U., **[Petersen, S.]{.underline}**, Schmidt, S., Wolf, G.,
    Jahnke, V., Dietel, M., and Petersen, I.: Patterns of chromosomal
    alterations in metastasizing and nonmetastasizing primary head and
    neck carcinomas. *Cancer Research.* 57: 5213-5216, 1997.

2.  Petersen, I., Bujard, M., **[Petersen, S.]{.underline}**, Wolf, G.,
    Goeze, A., Schwendel, A., Langreck, H., Gellert, K., Reichel, M.,
    Just, K., du Manoir, S., Cremer, T., Dietel, M., and Ried, T.:
    Patterns of chromosomal imbalances in adenocarcinoma and squamous
    cell carcinoma of the lung. *Cancer Research.* 57: 2331-2335, 1997.

3.  **[Petersen, S.]{.underline}**, Rudolf, J., Bockmuhl, U., Gellert,
    K., Wolf, G., Dietel, M., and Petersen, I.: Distinct regions of
    allelic imbalance on chromosome 10q22-q26 in squamous cell
    carcinomas of the lung. *Oncogene.* 17: 449-454, 1998.

4.  **[Petersen, S.]{.underline}**, Saretzki, G., and von Zglinicki, T.:
    Preferential accumulation of single-stranded regions in telomeres of
    human fibroblasts. *Experimental Cell Research.* 239: 152-160, 1998.

5.  **[Petersen, S.]{.underline}**, Wolf, G., Bockmuhl, U., Gellert, K.,
    Dietel, M., and Petersen, I.: Allelic loss on chromosome 10q in
    human lung cancer: association with tumour progression and
    metastatic phenotype. *British Journal of Cancer.* 77: 270-276,
    1998.

6.  Petersen, I., Hidalgo, A., **[Petersen, S.]{.underline}**, Schluns,
    K., Schewe, C., Pacyna-Gengelbach, M., Goeze, A., Krebber, B.,
    Knosel, T., Kaufmann, O., Szymas, J., and von Deimling, A.:
    Chromosomal imbalances in brain metastases of solid tumors. *Brain
    Pathology.* 10: 395-401, 2000.

7.  Bockmuhl, U., Schluns, K., Kuchler, I., **[Petersen,
    S.]{.underline}**, and Petersen, I.: Genetic imbalances with impact
    on survival in head and neck cancer patients. *American Journal of
    Pathology.* 157: 369-375, 2000.

8.  Bockmuhl, U., Schmidt, S., **[Petersen, S.]{.underline}**, and
    Petersen, I.: Deletion of chromosome 10q-a marker for metastasis of
    head-neck carcinomas? *Laryngorhinootologie.* 79: 81-85, 2000.

9.  Hidalgo, A., Schewe, C., **[Petersen, S.]{.underline}**, Salcedo,
    M., Gariglio, P., Schluns, K., Dietel, M., and Petersen, I.: Human
    papilloma virus status and chromosomal imbalances in primary
    cervical carcinomas and tumour cell lines. *European Journal of
    Cancer.* 36: 542-548, 2000.

10. **[Petersen, S.]{.underline}**, Aninat-Meyer, M., Schluns, K.,
    Gellert, K., Dietel, M., and Petersen, I.: Chromosomal alterations
    in the clonal evolution to the metastatic stage of squamous cell
    carcinomas of the lung. *British Journal of Cancer.* 82: 65-73,
    2000.

***Peer reviewed articles (Cont.):***

11. **[Petersen, S.]{.underline}**, Heckert, C., Rudolf, J., Schluns,
    K., Tchernitsa, O. I., Schafer, R., Dietel, M., and Petersen, I.:
    Gene expression profiling of advanced lung cancer. *International
    Journal of Cancer.* 86: 512-517, 2000.

12. **[Petersen, S.]{.underline}**, Rudolf, J., Bockmuhl, U.,
    Deutschmann, N., Dietel, M., and Petersen, I.: Analysis of the DMBT1
    gene in carcinomas of the respiratory tract. *International Journal
    of Cancer.* 88: 71-76, 2000.

13. Speicher, M. R., **[Petersen, S.]{.underline}**, Uhrig, S., Jentsch,
    I., Fauth, C., Eils, R., and Petersen, I:. Analysis of chromosomal
    alterations in non-small cell lung cancer by multiplex-FISH,
    comparative genomic hybridization, and multicolor bar coding.
    *Laboratory Investigation.* 80: 1031-1041, 2000.

14. Kristiansen, G., Yu, Y., **[Petersen, S.]{.underline}**, Kaufmann,
    O., Schluns, K., Dietel, M., and Petersen, I.: Overexpression of
    c-erbB2 protein correlates with disease-stage and chromosomal gain
    at the c-erbB2 locus in non-small cell lung cancer. *European
    Journal of Cancer.* 37: 1089-1095, 2001.

15. Stassar, M. J., Devitt, G., Brosius, M., Rinnab, L., Prang, J.,
    Schradin, T., Simon, J., **[Petersen, S.]{.underline}**,
    Kopp-Schneider, A., and Zoller, M.: Identification of human renal
    cell carcinoma associated genes by suppression subtractive
    hybridization. *British Journal of Cancer.* 85, 1372-82, 2001.

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16. Garber, M. E., Troyanskaya, O.G., Schluens, K., **[Petersen,
    S.]{.underline}**, Thaesler, Z., Pacyna-Gengelbach, M., van de Rijn,
    M., Rosen, G. D., Perou, C. M., Whyte, R. I., Altman, R. B.,
    Brown, P. O., Botstein, D., and Petersen, I.: Diversity of gene
    expression in adenocarcinoma of the lung. *Proceedings of the
    National Academy of Science USA.* 98, 13784-9, 2001.

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17. Zhu, J., **[Petersen, S.]{.underline}**, Tessarollo, L., and
    Nussenzweig, A.: Targeted disruption of the Nijmegen breakage
    syndrome gene NBS1 leads to early embryonic lethality in mice.
    *Current Biology.* 11, 105-9, 2001.

18. **[Petersen, S.]{.underline}**, Casellas, R., Reina-San-Martin, B.,
    Chen, H.T., Difilippantonio, M.J., Wilson, P.C., Hanitsch, L.,
    Celeste, A., Muramatsu, M., Pilch, D.R., Redon, C., Ried, T.,
    Bonner, W.M., Honjo, T., Nussenzweig, M.C., and Nussenzweig, A.: AID
    is required to initiate Nbs1/gamma-H2AX focus formation and
    mutations at sites of class switching. *Nature.* 414: 660-665, 2001.

19. Goeze, A., Schluns, K., Wolf, G., Thasler, Z., **[Petersen,
    S.]{.underline}**, and Petersen, I.: Chromosomal imbalances of
    primary and metastatic lung adenocarcinomas. *Journal of Pathology.*
    196: 8-16, 2002.

***Peer reviewed articles (Cont.):***

20. Saretzki, G., **[Petersen, S.]{.underline}**, Petersen, I., Kolble,
    K., and von Zglinicki T.: hTERT gene dosage correlates with
    telomerase activity in human lung cancer cell lines. *Cancer
    Letters.* 176: 81-91, 2002.

21. Hauptmann, S., Denkert, C., Koch, I., **[Petersen,
    S.]{.underline}**, Schluns, K., Reles, A., Dietel, M. and Petersen,
    I.: Genetic alterations in epithelial ovarian tumors analyzed by
    comparative genomic hybridization. *Human Pathology.* 33: 632-641,
    2002.

22. Knosel, T., **[Petersen, S.]{.underline}**, Schwabe, H., Schluns,
    K., Stein, U., Schlag, P.M., Dietel, M. and Petersen, I.: Incidence
    of chromosomal imbalances in advanced colorectal carcinomas and
    their metastases. *Virchows Archive.* 440: 187-194, 2002.

23. Celeste, A., **[Petersen, S.]{.underline}**, Romanienko, P.J.,
    Fernandez-Capetillo, O., Chen, H.T., Sedelnikova, O.A.,
    Reina-San-Martin, B., Coppola, V., Meffre, E., Difilippantonio,
    M.J., Redon, C., Pilch, D.R., Olaru, A., Eckhaus, M.,
    Camerini-Otero, R.D., Tessarollo, L., Livak, F., Manova, K., Bonner,
    W.M., Nussenzweig, M.C. and Nussenzweig, A.: Genomic instability in
    mice lacking histone H2AX. *Science.* 296: 922-927, 2002.

24. Difilippantonio, M.J., **[Petersen, S.]{.underline}**, Chen, H.T.,
    Johnson, R., Jasin, M., Kanaar, R., Ried, T. and Nussenzweig, A.:
    Evidence for Replicative Repair of DNA Double-Strand Breaks Leading
    to Oncogenic Translocation and Gene Amplification. *Journal of
    Experimental Medicine.* 196: 469-480, 2002.

25. Chen, Y., **[Petersen, S.]{.underline}**, Pacyna-Gengelbach, M.,
    Pietas, A. and Petersen, I.: Identification of a novel
    homeobox-containing gene, LAGY, which is downregulated in lung
    cancer. *Oncology.* 64: 450-458, 2003.

26. **[Difilippantonio, S.]{.underline}**, Chen, Y., Pietas, A.,
    Schluns, K., Pacyna-Gengelbach, M., Deutschmann, N., Padilla-Nash,
    H.M., Ried, T. and Petersen, I.: Gene expression profiles in human
    non-small and small-cell lung cancers. *European Journal of Cancer.*
    39: 1936-1947, 2003.

27. Reina-San-Martin, B., **[Difilippantonio, S.]{.underline}**,
    Hanitsch, L., Masilamani, R.F., Nussenzweig, A. and Nussenzweig,
    M.C.: H2AX is required for recombination between Immunoglobulin
    Switch Regions but not for Intra-Switch Region recombination or
    Somatic Hypermutation. *Journal of Experimental Medicine.* 197:
    1767-1778, 2003.

28. Celeste, A.\*, **[Difilippantonio, S.]{.underline}**\*,
    Difilippantonio, M.J., Fernandez-Capetillo, O., Pilch, D.R.,
    Sedelnikova, O.A., Eckhaus, M., Ried, T., Bonner, W.M. and
    Nussenzweig, A.: H2AX haploinsufficiency modifies genomic stability
    and tumor susceptibility. *Cell.* 114: 371-383, 2003. \*These
    authors contributed equally to this work.

29. Ramiro, A.R., Jankovic, M., Eisenreich, T., **[Difilippantonio,
    S.]{.underline}**, Chen-Kiang, S., Muramatsu, M., Honjo, T.,
    Nussenzweig, A. and Nussenzweig, M.C.: AID is required for c-myc/IgH
    chromosome translocations in vivo. *Cell.* 118: 431-438, 2004.

***Peer reviewed articles (Cont.):***

30. Reina-San-Martin, B., Nussenzweig, M.C., Nussenzweig, A. and
    **[Difilippantonio, S.]{.underline}**: Genomic instability,
    endo-reduplication and diminished immunoglobulin class switch
    recombination in B cells lacking Nbs. *Proceedings of the National
    Academy of Science USA.* 102:1590-1595, 2005.

31. **[Difilippantonio, S.]{.underline}**, Celeste, A.,
    Fernandez-Capetillo, O., Chen, H., Reina-San-Martin, B., Van
    Laethem, F., Yang, Y.P., Petukhova, G.V., Eckhaus, M., Feigenbaum,
    L., Manova, K., Kruhlak, M., Camerini-Otero, R.D., Sharan, S.,
    Nussenzweig, M. and Nussenzweig, A.: Essential and dispensable roles
    of Nbs1 in the activation of the ATM kinase revealed in humanized
    mouse models. *Nature Cell Biology.* 7: 657-685, 2005.

32. Ward, I.M., **[Difilippantonio, S.]{.underline}**, Minn, K.,
    Mueller, M.D., Molina, J.R., Yu, X., Frisk, C.S., Ried, T.,
    Nussenzweig, A. and Chen, J.: 53BP1 cooperates with p53 and
    functions as a haploinsufficient tumor suppressor in mice.
    *Molecular and Cellular Biology.* 25: 10079-10086, 2005.

33. Ramiro, A.R., Jankovic, M., Callen, E., **[Difilippantonio,
    S.]{.underline}**, Chen, H., McBride, K.M., Eisenreich, T.R., Chen,
    J., Dickins, R.A., Lowe, S.W., Nussenzweig, A. and Nussenzweig,
    M.C.: Role of genomic instability and p53 in AID-induced c-myc-IgH
    translocations. *Nature*. 440:105-109, 2006.

34. Pellegrini, M., Celeste, A., [**Difilippantonio, S.**,]{.underline}
    Guo, R., Wang, W., Feigenbaum, L. and Nussenzweig, A.:
    Autophosphorylation at Serine 1987 is dispensable for murine Atm
    activation in vivo. *Nature*, 443: 222-225, 2006.

35. **[Difilippantonio, S.]{.underline}**, Celeste, A., Kruhlak, M.,
    Lee, Y., Difilippantonio, M.J., Feigenbaum, L., Jackson, S.P.,
    McKinnon, P.J. and Nussenzweig, A.: Distinct domains in Nbs1
    regulate irradiation-induced checkpoints and apoptosis. *Journal of
    Experimental Medicine,* 204: 1003-1011, 2007.

36. Callen, E., Jankovic, M., **[Difilippantonio, S.]{.underline}**,
    Daniel, J.A., Chen, H.T., Celeste, A., Pellegrini, M., McBride, K.,
    Wangsa, D., Bredemeyer, A., Sleckman, B.P., Ried, T., Nussenzweig,
    M.C. and Nussenzweig, A.: ATM prevents the persistence and
    propagation of chromosome breaks in lymphocytes. *Cell*, 130: 63-75,
    2007.

***Reviews:***

1.  Petersen, I., and **[Petersen, S.]{.underline}**: Towards a
    genetic-based classification of human lung cancer. *Analytical and
    Cellular Pathology* *22*, 111-21, 2001.

2.  **[Difilippantonio, S.]{.underline}** and Nussenzweig, A.: The
    NBS1-ATM connection revisited. *Cell Cycle*, 6: 2366-2370, 2007.

***Book Chapters:***

1.  **[Petersen, S.]{.underline}**, and Petersen, I. Expression
    profiling of lung cancer based on Suppression Subtraction
    Hybridization (SSH) in: Methods in Molecular Medicine, vol. 75: Lung
    Cancer, Vol. 2: Diagnostic and Therapeutic Methods and
    Reviews. (2002) Humana Press Inc., Totowa, Chapter 11, pg 189-207.