Center for Molecular Medicine Cologne

Hallek, Michael | Nguyen, Phuong-Hien | Rebollido-Rios, Rocio - A 04

Understanding the impact of the lymphoid microenvironment in B cell malignancies by single-cell analysis

Introduction

The application of targeted therapy and immunotherapy has significantly shifted cancer treatment towards a chemotherapy-free future. However, the emergence of resistance and the relatively low responsive rate remain the major challenges in these therapeutic options.

Based on the premise that the tumor microenvironment has a significant impact on cancer resistance to therapy, results from this project can be useful to design improved treatment regime for B cell malignancies.

Small molecule inhibitors targeting tyrosine kinases and immunotherapies including immune checkpoint blockade have tremendously improved the treatment of B lymphoid malignancies. Recently, increasing evidence emerged showing that the efficacy of these inhibitors depends not only on the inhibition of B cell-autonomous functions, but also by affecting several cell types in the tumor micrenvironment (TME). Using genetically engineered mouse models, we have discovered an unexpected, potent role of Lyn kinase for the formation of a pro-malignant lymphoid environment.

Moreover, we also observed a potential function of the immune checkpoint ligands in the TME contributing to lymphoma cell growth. Our preliminary analysis of the murine lymphoid organs revealed striking differences in the immune cell composition of the knockout mice compared to wild type counterpart, indicating a substantial contribution of these molecules in shaping a tumor-supportive niche. Therefore, the central aim of this project is to provide mechanistic insights into the highly complex and dynamic interactions between malignant cells and their TME, with single-cell analysis as the central technology.

Using the different knockout mouse lines with distinct defects in the immune microenvironment, we will develop bioinformatic framework for identification of the cellular composition and comparison of the immune transcriptional landscape in different disease stages, thereby providing a deeper understanding of the dialog that malignant cells and the TME can adopt during B cell lymphoma development.

Previous Work

In recent years, tremendous advances regarding the therapy of B lymphoid malignancies have been made with the development of small molecule kinase inhibitors targeting the B cell receptor (BCR)-associated kinases. Strikingly,increasing evidence emerged showing that the efficacy of these inhibitors depends not only on the inhibition of B cell-autonomous functions, but also by affecting several cell types in the TME.

This is strongly supported by clinical observations, because CLL patients treated with these kinase inhibitors showed a long-lasting lymphocytosis following the rapid decrease of lymphadenopathy, indicating that leukemic cells lose their contact to the TME and undergo a relocation from lymphoid homing organs to the periphery (Figure 1).

  1. Fischer K, Al-Sawaf O, Bahlo J, Fink AM, Tandon M, Dixon M, Robrecht S, Warburton S, Humphrey K, Samoylova O, Liberati AM, Pinilla-Ibarz J, Opat S, Sivcheva L, Le Dû K, Fogliatto LM, Niemann CU, Weinkove R, Robinson S, Kipps TJ, Boettcher S, Tausch E, Humerickhouse R, Eichhorst B, Wendtner CM, Langerak AW, Kreuzer KA, Ritgen M, Goede V, Stilgenbauer S, Mobasher M, and Hallek, M. (2019). Venetoclax and Obinutuzumab in Patients with CLL and Coexisting Conditions. N Engl J Med. 380, 2225-2236.
  2. Nguyen P.H.*, Niesen E.*, and Hallek M.: New roles for B cell receptor associated kinases: when the B cell is not the target. Leukemia2019 Mar; 33(3):576-587
  3. Herling, C.D., Abedpour, N., Weiss, J., Schmitt, A., Jachimowicz, R.D., Merkel, O., Cartolano, M., Oberbeck, S., Mayer, P., Berg, V., Thomalla, D., Kutsch, N., Stiefelhagen, M., Cramer, P., Wendtner, C.M., Persigehl, T., Saleh, A., Altmuller, J., Nurnberg, P., Pallasch, C., Achter, V., Lang, U., Eichhorst, B., Castiglione, R., Schafer, S.C., Buttner, R., Kreuzer, K.A., Reinhardt, H.C., Hallek, M., Frenzel, L.P., and Peifer, M. (2018). Clonal dynamics towards the development of venetoclax resistance in chronic lymphocytic leukemia. Nat Commun 9, 727-730.
  4. Nguyen, P.H., Fedorchenko, O., Rosen, N., Koch, M., Barthel, R., Winarski, T., Florin, A., Wunderlich, F.T., Reinart, N., and Hallek, M. (2016). LYN Kinase in the Tumor Microenvironment Is Essential for the Progression of Chronic Lymphocytic Leukemia. Cancer Cell 30, 610-622.
  5. Landau, D.A., Tausch, E., Taylor-Weiner, A.N., Stewart, C., Reiter, J.G., Bahlo, J., Kluth, S., Bozic, I., Lawrence, M., Bottcher, S., Carter, S.L., Cibulskis, K., Mertens, D., Sougnez, C.L., Rosenberg, M., Hess, J.M., Edelmann, J., Kless, S., Kneba, M., Ritgen, M., Fink, A., Fischer, K., Gabriel, S., Lander, E.S., Nowak, M.A., Dohner, H., Hallek, M.,* Neuberg, D.,* Getz, G.,* Stilgenbauer, S.,* and Wu, C.J.* (2015). Mutations driving CLL and their evolution in progression and relapse. Nature 526, 525-530. *Joint senior authors
  6. Reinart, N., Nguyen, P.H., Boucas, J., Rosen, N., Kvasnicka, H.M., Heukamp, L., Rudolph, C., Ristovska, V., Velmans, T., Mueller, C., Reiners, K.S., von Strandmann, E.P., Krause, G., Montesinos-Rongen, M., Schlegelberger, B., Herling, M., Hallek, M., and Fingerle-Rowson, G. (2013). Delayed development of chronic lymphocytic leukemia in the absence of macrophage migration inhibitory factor. Blood  121, 812-821.
  7. Hallek, M., Fischer, K., Fingerle-Rowson, G., Fink, A.M., Busch, R., Mayer, J., Hensel, M., Hopfinger, G., Hess, G., von Grunhagen, U., Bergmann, M., Catalano, J., Zinzani, P.L., Caligaris-Cappio, F., Seymour, J.F., Berrebi, A., Jager, U., Cazin, B., Trneny, M., Westermann, A., Wendtner, C.M., Eichhorst, B.F., Staib, P., Buhler, A., Winkler, D., Zenz, T., Bottcher, S., Ritgen, M., Mendila, M., Kneba, M., Dohner, H., and Stilgenbauer, S. (2010). Addition of rituximab to fludarabine and cyclophosphamide in patients with chronic lymphocytic leukaemia: a randomised, open-label, phase 3 trial. Lancet 376, 1164-1174.
  • Pacholewska A, Grimm C, Herling CD, Lienhard M, Konigs A, Timmermann B, Altmuller J, Mucke O, Reinhardt HC, Plass C, Herwig R, Hallek M, and Schweiger MR (2021). Altered DNA Methylation Profiles in SF3B1 Mutated CLL Patients. Int J Mol Sci22. doi:10.3390/ijms22179337.
                              • Lobastova L, Lettau M, Babatz F, de Oliveira TD, Nguyen PH, Pauletti BA, Schauss AC, Durkop H, Janssen O, Paes Leme AF, Hallek M, and Hansen HP (2021). CD30-Positive Extracellular Vesicles Enable the Targeting of CD30-Negative DLBCL Cells by the CD30 Antibody-Drug Conjugate Brentuximab Vedotin. Front Cell Dev Biol9, 698503. doi:10.3389/fcell.2021.698503.
                                                                            • Theobald SJ, Simonis A, Georgomanolis T, Kreer C, Zehner M, Eisfeld HS, Albert MC, Chhen J, Motameny S, Erger F, Fischer J, Malin JJ, Grab J, Winter S, Pouikli A, David F, Boll B, Koehler P, Vanshylla K, Gruell H, Suarez I, Hallek M, Fatkenheuer G, Jung N, Cornely OA, Lehmann C, Tessarz P, Altmuller J, Nurnberg P, Kashkar H, Klein F, Koch M, and Rybniker J (2021). Long-lived macrophage reprogramming drives spike protein-mediated inflammasome activation in COVID-19. EMBO Mol Med13, e14150. doi:10.15252/emmm.202114150.
                                                                                                                                                                                          • Kohlhaas V, Blakemore SJ, Al-Maarri M, Nickel N, Pal M, Roth A, Hovelmeyer N, Schafer SC, Knittel G, Lohneis P, Nikolic M, Wiederstein JL, Franitza M, Georgomonolis T, Reinart N, Herling M, Herling C, Hartmann EM, Rosenwald A, Klapper W, Buttner R, Moia R, Rossi D, Boldorini R, Gaidano G, Frenzel LP, Reinhardt HC, Bruning JC, Hallek M, Kruger M, Peifer M, Pallasch CP, and Wunderlich FT (2021). Active Akt signaling triggers CLL toward Richter transformation via overactivation of Notch1. Blood137, 646-660. doi:10.1182/blood.2020005734.
                                                                                                                                                                                          • Kohlhas V, Hallek M*, Nguyen PH*.: Constitutive activation of Lyn kinase enhances BCR responsiveness, but not the development of CLL in Eµ-TCL1 mice. Blood Adv. 2020 Dec 22;4(24):6106-6116. doi: 10.1182/bloodadvances.2020002584. PMID: 33351104
                                                                                                                                                                                          • Koch M, Reinartz S, Saggau J, Knittel G, Rosen N, Fedorchenko O, Thelen L, Barthel R, Reinart N, Seeger-Nukpezah T, Reinhardt HC, Hallek M, Nguyen PH.: Meta-Analysis Reveals Significant Sex Differences in Chronic Lymphocytic Leukemia Progression in the Eµ-TCL1 Transgenic Mouse Model. Cancers (Basel) 2020 Jul 20;12(7):1980. doi: 10.3390/cancers12071980.
                                                                                                                                                                                          • Barbarino V, Henschke S, Blakemore SJ, Izquierdo E, Michalik M, Nickel N, Mollenkotte I, Vorholt D, Muller L, Brinker R, Fedorchenko O, Mikhael N, Seeger-Nukpezah T, Hallek M, and Pallasch CP (2020). Macrophage-Mediated Antibody Dependent Effector Function in Aggressive B-Cell Lymphoma Treatment is Enhanced by Ibrutinib via Inhibition of JAK2. Cancers (Basel) 12.
                                                                                                                                                                                          • Barthel R, Fedorchenko O, Velmans T, Rosen N, Nguyen PH, Reinart N, Florin A, Herling M, Hallek M, Fingerle-Rowson G.: CD74 is dispensable for development of chronic lymphocytic leukemia in Eµ-TCL1 transgenic mice. Leuk Lymphoma. 2020 Jul 15:1-12. doi: 10.1080/10428194.2020.1791851. Online ahead of print. PMID: 32667245
                                                                                                                                                                                          • Braun F, Rinschen M, Buchner D, Bohl K, Plagmann I, Bachurski D, Richard Spath M, Antczak P, Gobel H, Klein C, Lackmann JW, Kretz O, Puelles VG, Wahba R, Hallek M, Schermer B, Benzing T, Huber TB, Beyer A, Stippel D, Kurschat CE, and Muller RU (2020). The proteomic landscape of small urinary extracellular vesicles during kidney transplantation. Journal of extracellular vesicles 10, e12026.
                                                                                                                                                                                          • Volz C, Breid S, Selenz C, Zaplatina A, Golfmann K, Meder L, Dietlein F, Borchmann S, Chatterjee S, Siobal M, Schottle J, Florin A, Koker M, Nill M, Ozretic L, Uhlenbrock N, Smith S, Buttner R, Miao H, Wang B, Reinhardt HC, Rauh D, Hallek M, Acker-Palmer A, Heukamp LC, and Ullrich RT (2020). Inhibition of Tumor VEGFR2 Induces Serine 897 EphA2-Dependent Tumor Cell Invasion and Metastasis in NSCLC. Cell Rep 31, 107568.
                                                                                                                                                                                          • Hansen HP, Paes Leme AF, and Hallek M (2020). Role of ADAM10 as a CD30 Sheddase in Classical Hodgkin Lymphoma. Front Immunol 11, 398.
                                                                                                                                                                                          • Di Cristanziano V, Meyer-Schwickerath C, Eberhardt KA, Rybniker J, Heger E, Knops E, Hallek M, Klein F, Holtick U, and Jung N (2020). Detection of SARS-CoV-2 viremia before onset of COVID-19 symptoms in an allo-transplanted patient with acute leukemia. Bone marrow transplantation 10.1038/s41409-020-01059-y.
                                                                                                                                                                                          • Kreuzer KA, Furman RR, Stilgenbauer S, Dubowy RL, Kim Y, Munugalavadla V, Lilienweiss E, Reinhardt HC, Cramer P, Eichhorst B, Hillmen P, O'Brien SM, Pettitt AR, and Hallek M (2020). The impact of complex karyotype on the overall survival of patients with relapsed chronic lymphocytic leukemia treated with idelalisib plus rituximab. Leukemia 34, 296-300.
                                                                                                                                                                                          • Oberbeck S, Schrader A, Warner K, Jungherz D, Crispatzu G, von Jan J, Chmielewski M, Ianevski A, Diebner HH, Mayer P, Kondo Ados A, Wahnschaffe L, Braun T, Muller TA, Wagle P, Bouska A, Neumann T, Putzer S, Varghese L, Pflug N, Thelen M, Makalowski J, Riet N, Gox HJM, Rappl G, Altmuller J, Kotrova M, Persigehl T, Hopfinger G, Hansmann ML, Schlosser H, Stilgenbauer S, Durig J, Mougiakakos D, von Bergwelt-Baildon M, Roeder I, Hartmann S, Hallek M, Moriggl RH, Bruggemann M, Aittokallio T, Iqbal J, Newrzela S, Abken H, and Herling M (2020). Non-canonical effector functions of the T-memory-like T-PLL cell are shaped by cooperative TCL1A and TCR signaling. Blood 10.1182/blood.2019003348.
                                                                                                                                                                                          Prof. Dr. Michael Hallek CMMC Cologne
                                                                                                                                                                                          Prof. Dr. Michael Hallek

                                                                                                                                                                                          Clinic I of Internal Medicine

                                                                                                                                                                                          CMMC - PI - A 04

                                                                                                                                                                                          Executive Board Member

                                                                                                                                                                                          Publications - Michael Hallek

                                                                                                                                                                                          Link to PubMed

                                                                                                                                                                                          Dr. Phuong-Hien Nguyen CMMC Cologne
                                                                                                                                                                                          Dr. Phuong-Hien Nguyen

                                                                                                                                                                                          Clinic I for Internal Medicine

                                                                                                                                                                                          CMMC - PI - CAP 19

                                                                                                                                                                                          CMMC - Co-PI - A 04

                                                                                                                                                                                          Publications - Phuong-Hien Nguyen

                                                                                                                                                                                          Link to PubMed

                                                                                                                                                                                          Dr. Rocio Rebollido-Rios CMMC Cologne
                                                                                                                                                                                          Dr. Rocio Rebollido-Rios

                                                                                                                                                                                          Clinic I for Internal Medicine

                                                                                                                                                                                          CMMC - Co-PI - A 04

                                                                                                                                                                                          Publications - Rocio Rebollido-Rios

                                                                                                                                                                                          Link to PubMed

                                                                                                                                                                                          Figure 1
                                                                                                                                                                                          Group Members

                                                                                                                                                                                          Nicole Jörns, Admin. Assistenz
                                                                                                                                                                                          Dr. rer. nat. Phuong-Hien Nguyen, Co-PI
                                                                                                                                                                                          Dr. rer. nat. Rocio Rebollido-Rios, Co-PI
                                                                                                                                                                                          Dr. rer. nat. Oleg Fedorchenko
                                                                                                                                                                                          PD. Dr. rer. nat. Hinrich Hansen
                                                                                                                                                                                          Dr. nat. med. cand. Daniel Bachurski
                                                                                                                                                                                          Dr. rer. nat. cand. Maximilian Koch
                                                                                                                                                                                          Dr. rer. nat. cand. Sebastian Reinartz
                                                                                                                                                                                          Dr. rer. nat. cand. Viktoria Kohlhaas
                                                                                                                                                                                          Dr. rer. nat. cand. Thais Dolzany De Oliveira
                                                                                                                                                                                          Dr. rer. nat. cand. Maria José Fariña Morillas
                                                                                                                                                                                          Carina Folger, TA
                                                                                                                                                                                          Natascha Rosen, TA
                                                                                                                                                                                          Hedwig Göx, Medical Student
                                                                                                                                                                                          Lea Reemann, Medical Student
                                                                                                                                                                                          Alexander vom Stein, Medical Student
                                                                                                                                                                                          Max Bondarchenko, Master Student
                                                                                                                                                                                          Ludmila Lobastova, Master Student
                                                                                                                                                                                          Julia Saggau, Master Student