SFB 684:  Molecular Mechnisms of Normal and Malignant Hematopoiesis

• Subject Area: Medicine ; Biology
• Term: from 2006 to 2014
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 13759457

• Final Report Year: 2015

Final Report Abstract

Malignant diseases constitute one of the most challenging health problems in our society. In parallel to the increasing life expectancy, the number of patients suffering from cancer is steadily increasing. To cope with this development, prevention and early detection as well as more effective therapies are urgently needed. Among the malignant disorders, leukemias serve as model diseases for which basic principles of pathogenesis and therapy of cancer were first explored. The collaborative research center (CRC) 684 “Molecular Mechanisms of Normal and Malignant Hematopoiesis” was established to dissect the molecular pathways and mechanisms that govern normal and malignant hematopoiesis. This was accomplished by a close interaction between investigations focusing on single molecules or defined molecular pathways and analyses addressing the complexity of the disease and the physiological differentiation processes by using sophisticated model systems and patient material. The basis for this challenging approach was provided through the cooperation between well established basic researchers, junior group leaders and physician scientists and was supported by the ready access to patient material from the controlled clinical trials of the German AML Cooperative Group. The CRC focused on mechanisms of cellular growth, transcriptional regulation and signal transduction. Furthermore, projects analyzed chromatin structure, the influence of genetic and epigenetic processes on genome stability, normal hematopoietic development and developed animal models for both myeloid and lymphoid malignancies. Scientific highlights from the first funding period include novel technical developments and interesting biological insights. Using continuous monitoring of single cells, T. Schroeder could show that endothelial cells give rise to blood cells. This work solved a long standing mystery about the identity of the cells that generate the first blood cells in the mammalian embryo. The same technique allowed T. Schroeder to demonstrate that hematopoietic cytokines play an instructive – and not a permissive – role in lineage choice. H. Leonhardt also pioneered a new method of light microscopy which provides resolutions beyond the Abbe limit. With this so-called three-dimensional structured illumination microscopy (3D-SIM) his group was able to resolve for example individual nuclear pore complexes in the nuclear membrane. This technology had great impact on the analysis of cellular structures such as the nucleus. The careful analysis of a specific leukemia fusion gene, CALM-AF10 in a bone marrow transplantation model led to the identification of a subpopulation of leukemic cells that were enriched in leukemia propagating cell. This work in C. Buske's and S. Bohlander's group gave new insights into the biology of leukemic stem cells. In the second funding period P. Greif and A. Dufour applied new DNA sequencing techniques and functional assays to identify somatic GATA2 zinc finger 1 (ZF1) mutations in patients with biallelic CEBPA gene mutations. This work from S. Bohlander´s and K. Spiekermann´s group provides evidence for a genetically distinct subgroup of Acute Myeloid Leukemia. K.-P. Hopfner´s group determined the crystal structure of the catalytic head of the Mre11 nuclease and Rad50 ABC ATPase (MR) complex and analyzed ATP-dependent conformational changes. They could show that MR is an ATPcontrolled transient molecular clamp at DNA double-strand breaks. J. Ruland´s group could show that the fusion protein ITK-SYK associates constitutively with lipid rafts in T cells and triggers antigen-independent phosphorylation of T cell receptor (TCR)- proximal proteins. This model can serve as a robust clinically relevant and genetically tractable model of human peripheral T cell lymphomas. H. Leonhardt´s group worked out that the regulation of the levels of 5-methylcytosine (mC) and its oxidized derivatives are essential for normal cell homeostasis and identified the readers, writers, and erasers of these marks. In a second work, lamin B receptor (Lbr) and lamin A (Lmna) genes that encode nuclear envelope (NE) proteins were analyzed. This elegant work could show how changes in NE composition contribute to regulating heterochromatin positioning, gene expression, and cellular differentiation during development. D. Eick´s group contributed two aspects to the general mechanisms of regulation of carboxyterminal domain (CTD) of RNA polymerase II (RNAPII). It is very likely that the mechanisms studied in this CRC will also be relevant to other types of cancer (e.g. the concept of the “cancer stem cell” or common “oncogenic pathways”). It has become clear in the first and second funding period of this CRC that a multitude of innovative collaborations between the different members of the CRCs have been initiated and successfully completed. Many of these collaborations would not have been possible without the formal frame work which was provided by this collaborative research center.

Publications

• 2006. Acute myeloid leukemia is propagated by a leukemic stem cell with lymphoid characteristics in a mouse model of CALM/AF10-positive leukemia. Cancer Cell. 10:363-374
  Deshpande, A.J., M. Cusan, V.P. Rawat, H. Reuter, A. Krause, C. Pott, L. Quintanilla-Martinez, P. Kakadia, F. Kuchenbauer, F. Ahmed, E. Delabesse, M. Hahn, P. Lichter, M. Kneba, W. Hiddemann, E. Macintyre, C. Mecucci, W.D. Ludwig, R.K. Humphries, S.K. Bohlander, M. Feuring-Buske, and C. Buske
  
• 2006. Targeting and tracing antigens in live cells with fluorescent nanobodies. Nat Methods. 3:887-889
  Rothbauer, U., K. Zolghadr, S. Tillib, D. Nowak, L. Schermelleh, A. Gahl, N. Backmann, K. Conrath, S. Muyldermans, M.C. Cardoso, and H. Leonhardt
  
• 2008. Overexpression of CDX2 perturbs HOX gene expression in murine progenitors depending on its N-terminal domain and is closely correlated with deregulated HOX gene expression in human acute myeloid leukemia. Blood. 111:309-319
  Rawat, V.P., S. Thoene, V.M. Naidu, N. Arseni, B. Heilmeier, K. Metzeler, K. Petropoulos, A. Deshpande, Quintanilla-Martinez, S.K. Bohlander, K. Spiekermann, W. Hiddemann, M. Feuring-Buske, and C. Buske
  
• 2008. Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy. Science. 320:1332-1336
  Schermelleh, L., P.M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M.C. Cardoso, D.A. Agard, M.G. Gustafsson, H. Leonhardt, and J.W. Sedat
  
• 2008. The leukemogenic CALM/AF10 fusion protein alters the subcellular localization of the lymphoid regulator Ikaros. Oncogene. 27:2886-2896
  Greif, P.A., B. Tizazu, A. Krause, E. Kremmer, and S.K. Bohlander
  
• 2009. CBL exon 8/9 mutants activate the FLT3 pathway and cluster in core binding factor/11q deletion acute myeloid leukemia/myelodysplastic syndrome subtypes. Clinical cancer research : an official journal of the American Association for Cancer Research. 15:2238-2247
  Reindl, C., H. Quentmeier, K. Petropoulos, P.A. Greif, T. Benthaus, B. Argiropoulos, G. Mellert, S. Vempati, J. Duyster, C. Buske, S.K. Bohlander, K.R. Humphries, W. Hiddemann, and K. Spiekermann
  
• 2009. Continuous single-cell imaging of blood generation from haemogenic endothelium. Nature. 457:896-900
  Eilken, H.M., S. Nishikawa, and T. Schroeder
  
• 2009. Global reduction of the epigenetic H3K79 methylation mark and increased chromosomal instability in CALM-AF10-positive leukemias. Blood. 114:651-658
  Lin, Y.H., P.M. Kakadia, Y. Chen, Y.Q. Li, A.J. Deshpande, C. Buske, K.L. Zhang, Y. Zhang, G.L. Xu, and S.K. Bohlander
  
• 2009. Hematopoietic cytokines can instruct lineage choice. Science. 325:217-21
  Rieger, M.A., P.S. Hoppe, B.M. Smejkal, A.C. Eitelhuber, and T. Schroeder
  
• 2009. Notch1, Notch2, and Epstein-Barr virus-encoded nuclear antigen 2 signaling differentially affects proliferation and survival of Epstein-Barr virus-infected B cells. Blood. 113:5506-5515
  Kohlhof, H., F. Hampel, R. Hoffmann, H. Burtscher, U.H. Weidle, M. Holzel, D. Eick, U. Zimber-Strobl, and L.J. Strobl
  
• 2009. Nuclear architecture of rod photoreceptor cells adapts to vision in mammalian evolution. Cell. 137:356-368
  Solovei, I., M. Kreysing, C. Lanctot, S. Kosem, L. Peichl, T. Cremer, J. Guck, and B. Joffe
  
• 2009. Syk kinase signalling couples to the Nlrp3 inflammasome for anti-fungal host defence. Nature. 459:433-436
  Gross, O., H. Poeck, M. Bscheider, C. Dostert, N. Hannesschlager, S. Endres, G. Hartmann, A. Tardivel, E. Schweighoffer, V. Tybulewicz, A. Mocsai, J. Tschopp, and J. Ruland
  
• 2010. Acute myeloid leukemia with biallelic CEBPA gene mutations and normal karyotype represents a distinct genetic entity associated with a favorable clinical outcome. Journal of Clinical Oncology 28:570-577
  Dufour, A., F. Schneider, K.H. Metzeler, E. Hoster, S. Schneider, E. Zellmeier, T. Benthaus, M.C. Sauerland, W.E. Berdel, T. Buchner, B. Wormann, J. Braess, W. Hiddemann, S.K. Bohlander, and K. Spiekermann
  
• 2010. Oncogenic JAK2V617F requires an intact SH2-like domain for constitutive activation and induction of a myeloproliferative disease in mice. Blood. 116:4600-4611
  Gorantla, S.P., T.N. Dechow, R. Grundler, A.L. Illert, C.M. Zum Buschenfelde, M. Kremer, C. Peschel, and J. Duyster
  
• 2010. Recognition of RNA virus by RIG-I results in activation of CARD9 and inflammasome signaling for interleukin 1 beta production. Nat Immunol. 11:63-69
  Poeck, H., M. Bscheider, O. Gross, K. Finger, S. Roth, M. Rebsamen, N. Hannesschlager, M. Schlee, S. Rothenfusser, W. Barchet, H. Kato, S. Akira, S. Inoue, S. Endres, C. Peschel, G. Hartmann, V. Hornung, and J. Ruland
  
• 2010. The fusion kinase ITK-SYK mimics a T cell receptor signal and drives oncogenesis in conditional mouse models of peripheral T cell lymphoma. J Exp Med. 207:1031-1044
  Pechloff, K., J. Holch, U. Ferch, M. Schweneker, K. Brunner, M. Kremer, T. Sparwasser, L. Quintanilla- Martinez, U. Zimber-Strobl, B. Streubel, A. Gewies, C. Peschel, and J. Ruland
  
• 2011. A20 (TNFAIP3) deficiency in myeloid cells triggers erosive polyarthritis resembling rheumatoid arthritis. Nat Genet. 43:908-912
  Matmati, M., P. Jacques, J. Maelfait, E. Verheugen, M. Kool, M. Sze, L. Geboes, E. Louagie, C. Mc Guire, L. Vereecke, Y. Chu, L. Boon, S. Staelens, P. Matthys, B.N. Lambrecht, M. Schmidt- Supprian, M. Pasparakis, D. Elewaut, R. Beyaert, and G. van Loo
  
• 2011. An RNAibased system for loss-of-function analysis identifies Raf1 as a crucial mediator of BCR-ABL- driven leukemogenesis. Blood. 118:2200-2210
  Albers, C., A.L. Illert, C. Miething, H. Leischner, M. Thiede, C. Peschel, and J. Duyster
  
• 2011. B cells lacking the tumor suppressor TNFAIP3/A20 display impaired differentiation and hyperactivation and cause inflammation and autoimmunity in aged mice. Blood. 117:2227-2236
  Chu, Y., J.C. Vahl, D. Kumar, K. Heger, A. Bertossi, E. Wojtowicz, V. Soberon, D. Schenten, B. Mack, M. Reutelshofer, R. Beyaert, K. Amann, G. van Loo, and M. Schmidt-Supprian
  
• 2011. CD19-independent instruction of murine marginal zone B-cell development by constitutive Notch2 signaling. Blood. 118:6321-6331
  Hampel, F., S. Ehrenberg, C. Hojer, A. Draeseke, G. Marschall-Schroter, R. Kuhn, B. Mack, O. Gires, C.J. Vahl, M. Schmidt-Supprian, L.J. Strobl, and U. Zimber-Strobl
  
• 2011. Role for hACF1 in the G2/M damage checkpoint. Nucleic Acids Res. 39:8445- 8456
  Sanchez-Molina, S., O. Mortusewicz, B. Bieber, S. Auer, M. Eckey, H. Leonhardt, A.A. Friedl, and P.B. Becker
  
• 2011. The C-terminal domain of RNA polymerase II is modified by site-specific methylation. Science. 332:99-103
  Sims, R.J., 3rd, L.A. Rojas, D. Beck, R. Bonasio, R. Schuller, W.J. Drury, D. Eick, and D. Reinberg
  
• 2011. The Mre11:Rad50 structure shows an ATP-dependent molecular clamp in DNA double-strand break repair. Cell. 145:54-66
  Lammens, K., D.J. Bemeleit, C. Mockel, E. Clausing, A. Schele, S. Hartung, C.B. Schiller, M. Lucas, C. Angermuller, J. Soding, K. Strasser, and K.P. Hopfner
  
• 2011. The SUMO system controls nucleolar partitioning of a novel mammalian ribosome biogenesis complex. EMBO J. 30:1067-1078
  Finkbeiner, E., M. Haindl, and S. Muller
  
• 2012. Structure of Mre11-Nbs1 complex yields insights into ataxia-telangiectasia-like disease mutations and DNA damage signaling. Nat Struct Mol Biol. 19:693-700
  Schiller, C.B., K. Lammens, I. Guerini, B. Coordes, H. Feldmann, F. Schlauderer, C. Mockel, A. Schele, K. Strasser, S.P. Jackson, and K.P. Hopfner
  
• 2012. CTD tyrosine phosphorylation impairs termination factor recruitment to RNA polymerase II. Science. 336:1723-1725
  Mayer, A., M. Heidemann, M. Lidschreiber, A. Schreieck, M. Sun, C. Hintermair, E. Kremmer, D. Eick, and P. Cramer
  (See online at https://doi.org/10.1126/science.1219651)
• 2012. GATA2 zinc finger 1 mutations associated with biallelic CEBPA mutations define a unique genetic entity of acute myeloid leukemia. Blood. 120:395-403
  Greif, P.A., A. Dufour, N.P. Konstandin, B. Ksienzyk, E. Zellmeier, B. Tizazu, J. Sturm, T. Benthaus, T. Herold, M. Yaghmaie, P. Dorge, K.P. Hopfner, A. Hauser, A. Graf, S. Krebs, H. Blum, P.M. Kakadia, S. Schneider, E. Hoster, F. Schneider, M. Stanulla, J. Braess, M.C. Sauerland, W.E. Berdel, T. Buchner, B.J. Woermann, W. Hiddemann, K. Spiekermann, and S.K. Bohlander
  (See online at https://doi.org/10.1182/blood-2012-01-403220)
• 2012. In vivo imaging enables high resolution preclinical trials on patients' leukemia cells growing in mice. PloS one. 7:e52798
  Terziyska, N., C. Castro Alves, V. Groiss, K. Schneider, K. Farkasova, M. Ogris, E. Wagner, H. Ehrhardt, R.J. Brentjens, U. zur Stadt, M. Horstmann, L. Quintanilla-Martinez, and I. Jeremias
  (See online at https://doi.org/10.1371/journal.pone.0052798)
• 2012. Leukemia-initiating cells of patient-derived acute lymphoblastic leukemia xenografts are sensitive toward TRAIL. Blood. 119:4224-4227
  Castro Alves, C., N. Terziyska, M. Grunert, S. Gundisch, U. Graubner, L. Quintanilla-Martinez, and I. Jeremias
  (See online at https://doi.org/10.1182/blood-2011-08-370114)
• 2012. SRC is a signaling mediator in FLT3-ITD- but not in FLT3-TKD-positive AML. Blood. 119:4026-4033
  Leischner, H., C. Albers, R. Grundler, E. Razumovskaya, K. Spiekermann, S. Bohlander, L. Ronnstrand, K. Gotze, C. Peschel, and J. Duyster
  (See online at https://doi.org/10.1182/blood-2011-07-365726)
• 2012. The FLT3ITD mRNA level has a high prognostic impact in NPM1 mutated, but not in NPM1 unmutated, AML with a normal karyotype. Blood. 119:4383-4386
  Schneider, F., E. Hoster, M. Unterhalt, S. Schneider, A. Dufour, T. Benthaus, G. Mellert, E. Zellmeier, P.M. Kakadia, S.K. Bohlander, M. Feuring-Buske, C. Buske, J. Braess, A. Heinecke, M.C. Sauerland, W.E. Berdel, T. Buchner, B.J. Wormann, W. Hiddemann, and K. Spiekermann
  
• 2012. The germinal center kinase TNIK is required for canonical NF-kappaB and JNK signaling in B-cells by the EBV oncoprotein LMP1 and the CD40 receptor. PLoS Biol. 10:e1001376
  Shkoda, A., J.A. Town, J. Griese, M. Romio, H. Sarioglu, T. Knofel, F. Giehler, and A. Kieser
  (See online at https://doi.org/10.1371/journal.pbio.1001376)
• 2013. Dynamic readers for 5-(hydroxy)methylcytosine and its oxidized derivatives. Cell. 152:1146-1159
  Spruijt, C.G., F. Gnerlich, A.H. Smits, T. Pfaffeneder, P.W. Jansen, C. Bauer, M. Munzel, M. Wagner, M. Muller, F. Khan, H.C. Eberl, A. Mensinga, A.B. Brinkman, K. Lephikov, U. Muller, J. Walter, R. Boelens, H. van Ingen, H. Leonhardt, T. Carell, and M. Vermeulen
  (See online at https://doi.org/10.1016/j.cell.2013.02.004)
• 2013. Exome sequencing identifies recurring FLT3 N676K mutations in core-binding factor leukemia. Blood. 122:1761-1769
  Opatz, S., H. Polzer, T. Herold, N.P. Konstandin, B. Ksienzyk, E. Zellmeier, S. Vosberg, A. Graf, S. Krebs, H. Blum, K.P. Hopfner, P.M. Kakadia, S. Schneider, A. Dufour, J. Braess, M.C. Sauerland, W.E. Berdel, T. Buchner, B.J. Woermann, W. Hiddemann, K. Spiekermann, S.K. Bohlander, and P.A. Greif
  (See online at https://doi.org/10.1182/blood-2013-01-476473)
• 2013. LBR and lamin A/C sequentially tether peripheral heterochromatin and inversely regulate differentiation. Cell. 152:584-598
  Solovei, I., A.S. Wang, K. Thanisch, C.S. Schmidt, S. Krebs, M. Zwerger, T.V. Cohen, D. Devys, R. Foisner, L. Peichl, H. Herrmann, H. Blum, D. Engelkamp, C.L. Stewart, H. Leonhardt, and B. Joffe
  
• 2013. Suv4-20h2 mediates chromatin compaction and is important for cohesin recruitment to heterochromatin. Genes Dev. 27:859-872
  Hahn, M., S. Dambacher, S. Dulev, A.Y. Kuznetsova, S. Eck, S. Worz, D. Sadic, M. Schulte, J.P. Mallm, A. Maiser, P. Debs, H. von Melchner, H. Leonhardt, L. Schermelleh, K. Rohr, K. Rippe, Z. Storchova, and G. Schotta
  (See online at https://doi.org/10.1101/gad.210377.112)
• 2014. B-cell expansion and lymphomagenesis induced by chronic CD40 signaling is strictly dependent on CD19. Cancer Res. 74:4318-4328
  Hojer, C., S. Frankenberger, L.J. Strobl, S. Feicht, K. Djermanovic, F. Jagdhuber, C. Homig-Holzel, U. Ferch, J. Ruland, K. Rajewsky, and U. Zimber-Strobl
  (See online at https://doi.org/10.1158/0008-5472.CAN-13-3274)
• 2014. Clec12a is an inhibitory receptor for uric acid crystals that regulates inflammation in response to cell death. Immunity. 40:389-399
  Neumann, K., M. Castineiras-Vilarino, U. Hockendorf, N. Hannesschlager, S. Lemeer, D. Kupka, S. Meyermann, M. Lech, H.J. Anders, B. Kuster, D.H. Busch, A. Gewies, R. Naumann, O. Gross, and J. Ruland
  (See online at https://doi.org/10.1016/j.immuni.2013.12.015)
• 2014. Rad50- CARD9 interactions link cytosolic DNA sensing to IL-1beta production. Nat Immunol. 15:538-545
  Roth, S., A. Rottach, A.S. Lotz-Havla, V. Laux, A. Muschaweckh, S.W. Gersting, A.C. Muntau, K.P. Hopfner, L. Jin, K. Vanness, J.H. Petrini, I. Drexler, H. Leonhardt, and J. Ruland
  (See online at https://doi.org/10.1038/ni.2888)
• 2014. Structure of the Rad50 DNA double-strand break repair protein in complex with DNA. EMBO J. 33:2847-2859
  Rojowska, A., K. Lammens, F.U. Seifert, C. Direnberger, H. Feldmann, and K.P. Hopfner
  (See online at https://doi.org/10.15252/embj.201488889)