CMMC: Hallek, Michael | Nguyen, Phuong-Hien | Rebollido-Rios, Rocio

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.

Kinase inhibitors are now routinely used for treating hematological malignancies. The modulation of non-malignant bystander cells is not only an apparent off-target effect of these inhibitors, but also contributes importantly to the mechanisms of action of kinase inhibitors in B cell leukemia. Using genetically engineered mouse models, we have discovered an unexpected, potent role of Lyn kinase for the formation of a pro-malignant lymphoid environment. We could show that deletion of Lyn resulted in significantly reduced lymphoma burden in vivo using two validated transgenic mouse models for B cell tumors, which was due to the loss of Lyn and its substrate Btk in the tumor microenvironment (TME) rather than within the leukemic cells. Lyn was shown to have important functions in numerous hematopoietic cells of the myeloid lineage. Particularly, increasing evidence indicates that Lyn is involved in regulating the function of various cell types within the non-hematopoietic compartment. Using a comprehensive multi-omics profiling approach, we found that the presence of LYN kinase in the stromal microenvironment supports leukemic cell survival by shaping the cancer-associated-fibroblast (CAF)-like polarization of stromal cells and maintaining a tumor supportive extracellular matrix (ECM) composition. Deletion of LYN in stromal cells consistently decreased the capacity of stromal cells to support leukemic cell survival.

Our project provides mechanistic insights into the highly complex and dynamic interactions between malignant cells and their TME, with single-cell transcriptome as the central technology. Using the different knockout mouse lines with distinct defects in the immune microenvironment, we develop a bioinformatics framework to identify cellular populations and compare the immune transcriptional landscape at different disease stages, thereby providing a deeper understanding of the dialog that malignant cells and the TME can adopt during B cell lymphoma development. 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

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).

Figure 1

CMMC Research — Fig. 1. Roles of the BCR-associated kinases in B cell lymphomas and effects of their inhibition (Nguyen, Niesen & Hallek, Leukemia 2019).

Project Related Publications

Nguyen PH, Fedorchenko O, Rosen N, Koch M, Barthel R, Winarski T, Florin A, Wunderlich, FT, Reinart N, and Hallek M. LYN Kinase in the Tumor Microenvironment Is Essential for the Progression of Chronic Lymphocytic Leukemia. Cancer Cell 2016 30, 610-622.

Nguyen PH*, Niesen E*, and Hallek M. New roles for B cell receptor associated kinases: when the B cell is not the target. Leukemia 2019 Mar; 33(3):576-587

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.

Izquierdo E, Vorholt D, Blakemore S, Sackey B, Nolte JL, Barbarino V, Schmitz J, Nickel N, Bachurski D, Lobastova L, Nikolic M, Michalik M, Brinker R, Merkel O, Franitza M, Georgomanolis T, Neuhaus R, Koch M, Nasada N, Knittel G, Chapuy B, Ludwig N, Meese E, Frenzel L, Reinhardt HC, Peifer M, Rebollido-Rios R, Bruns H, Krüger M, Hallek M, Pallasch CP: Extracellular vesicles and PD-L1 suppress macrophages, inducing therapy resistance in TP53-deficient B-cell malignancies. Blood. 2022 Jun 23;139(25):3617-3629.

Rusyn L, Reinartz S, Nikiforov A, Mikhael N, Vom Stein A, Kohlhas V, Bloehdorn J, Stilgenbauer S, Lohneis P, Buettner R, Robrecht S, Fischer K, Pallasch C, Hallek M, Nguyen PH, Seeger-Nukpezah T: The scaffold protein NEDD9 is necessary for leukemia-cell migration and disease progression in a mouse model of chronic lymphocytic leukemia. Leukemia. 2022 Jul;36(7):1794-1805.

Selected Publications 2022

Cramer P, Fürstenau M, Robrecht S, Giza A, Zhang C, Fink AM, Fischer K, Langerbeins P, Al-Sawaf O, Tausch E, Schneider C, Schetelig J, Dreger P, Böttcher S, Kreuzer KA, Schilhabel A, Ritgen M, Brüggemann M, Kneba M, Stilgenbauer S, Eichhorst B, Hallek M. Obinutuzumab, acalabrutinib, and venetoclax, after an optional debulking with bendamustine in relapsed or refractory chronic lymphocytic leukaemia (CLL2-BAAG): a multicentre, open-label, phase 2 trial. Lancet Haematol. 2022 Aug 18:S2352-3026(22)00211-3. doi: 10.1016/S2352-3026(22)00211-3.
Knisbacher BA, Lin Z, Hahn CK, Nadeu F, Duran-Ferrer M, Stevenson KE, Tausch E, Delgado J, Barbera-Mourelle A, Taylor-Weiner A, Bousquets-Muñoz P, Diaz-Navarro A, Dunford A, Anand S, Kretzmer H, Gutierrez-Abril J, López-Tamargo S, Fernandes SM, Sun C, Sivina M, Rassenti LZ, Schneider C, Li S, Parida L, Meissner A, Aguet F, Burger JA, Wiestner A, Kipps TJ, Brown JR, Hallek M, Stewart C, Neuberg DS, Martín-Subero JI, Puente XS, Stilgenbauer S, Wu CJ, Campo E, Getz G. Molecular map of chronic lymphocytic leukemia and its impact on outcome. Nat Genet. 2022 Aug 4. doi: 10.1038/s41588-022-01140-w.
Theobald SJ, Simonis A, Mudler JM, Göbel U, Acton R, Kohlhas V, Albert MC, Hellmann AM, Malin JJ, Winter S, Hallek M, Walczak H, Nguyen PH, Koch M, Rybniker J. Spleen tyrosine kinase mediates innate and adaptive immune crosstalk in SARS-CoV-2 mRNA vaccination. EMBO Mol Med. 2022 Aug 8;14(8):e15888. doi: 10.15252/emmm.202215888.
von Tresckow J, Cramer P, Robrecht S, Langerbeins P, Fink AM, Al-Sawaf O, Fürstenau M, Illmer T, Klaproth H, Tausch E, Ritgen M, Fischer K, Wendtner CM, Kreuzer KA, Leukemia. 2022 Aug;36(8):2125-2128. doi: 10.1038/s41375-022-01629-7.Stilgenbauer S, Böttcher S, Eichhorst BF, Hallek M. Sequential treatment with bendamustine, obinutuzumab (GA101) and Ibrutinib in chronic lymphocytic leukemia (CLL): final results of the CLL2-BIG trial. Leukemia. 2022 Aug;36(8):2125-2128. doi: 10.1038/s41375-022-01629-7.
Thomalla D, Beckmann L, Grimm C, Oliverio M, Meder L, Herling CD, Nieper P, Feldmann T, Merkel O, Lorsy E, da Palma Guerreiro A, von Jan J, Kisis I, Wasserburger E, Claasen J, Faitschuk-Meyer E, Altmüller J, Nürnberg P, Yang TP, Lienhard M, Herwig R, Kreuzer KA, Pallasch CP, Buettner R, Schäfer SC, Hartley J, Abken H, Peifer M, Kashkar H, Knittel G, Eichhorst B, Ullrich RT, Herling M, Reinhardt HC, Hallek M, Schweiger MR, Frenzel LP.: Deregulation and epigenetic modification of BCL2-family genes cause resistance to venetoclax in hematologic malignancies. Blood. 2022 Jun 15:blood.2021014304. doi: 10.1182/blood.2021014304.
Kutsch N, Pallasch C, Decker T, Hebart H, Chow KU, Graeven U, Kisro J, Kroeber A, Tausch E, Fischer K, Fink AM, Wendtner CM, Ritgen M, Stilgenbauer S, Zhang D, Li B, Jürgensmeier JM, Rajakumaraswamy N, Bhargava P, Hallek M, Eichhorst B. Efficacy and Safety of Tirabrutinib and Idelalisib With or Without Obinutuzumab in Relapsed Chronic Lymphocytic Leukemia. Hemasphere. 2022 May 25;6(6):e729. doi: 10.1097/HS9.0000000000000729.
Langerbeins P, Zhang C, Robrecht S, Cramer P, Fürstenau M, Al-Sawaf O, von Tresckow J, Fink AM, Kreuzer KA, Vehling-Kaiser U, Tausch E, Müller L, Eckart MJ, Schlag R, Freier W, Gaska T, Balser C, Reiser M, Stauch M, Wendtner CM, Fischer K, Stilgenbauer S, Eichhorst B, Hallek M. The CLL12 trial: ibrutinib vs placebo in treatment-naïve, early-stage chronic lymphocytic leukemia. Blood. 2022 Jan 13;139(2):177-187. doi: 10.1182/blood.2021010845.

Selectet Publications 2021

Hallek M, Al-Sawaf O. Chronic lymphocytic leukemia: 2022 update on diagnostic and therapeutic procedures. Am J Hematol. 2021 Dec 1;96(12):1679-1705. doi: 10.1002/ajh.26367.

Bloehdorn J, Braun A, Taylor-Weiner A, Jebaraj BMC, Robrecht S, Krzykalla J, Pan H, Giza A, Akylzhanova G, Holzmann K, Scheffold A, Johnston HE, Yeh RF, Klymenko T, Tausch E, Eichhorst B, Bullinger L, Fischer K, Weisser M, Robak T, Schneider C, Gribben J, Dahal LN, Carter MJ, Elemento O, Landau DA, Neuberg DS, Cragg MS, Benner A, Hallek M, Wu CJ, Döhner H, Stilgenbauer S, Mertens D. Multi-platform profiling characterizes molecular subgroups and resistance networks in chronic lymphocytic leukemia. Nat Commun. 2021 Sep 13;12(1):5395. doi: 10.1038/s41467-021-25403-y.

Pacholewska A, Grimm C, Herling CD, Lienhard M, Königs A, Timmermann B, Altmüller J, Mücke O, Reinhardt HC, Plass C, Herwig R, Hallek M, Schweiger MR. Altered DNA Methylation Profiles in SF3B1 Mutated CLL Patients. Int J Mol Sci. 2021 Aug 28;22(17):9337. doi: 10.3390/ijms22179337.

Theobald SJ, Simonis A, Georgomanolis T, Kreer C, Zehner M, Eisfeld HS, Albert MC, Chhen J, Motameny S, Erger F, Fischer J, Malin JJ, Gräb J, Winter S, Pouikli A, David F, Böll B, Koehler P, Vanshylla K, Gruell H, Suárez I, Hallek M, Fätkenheuer G, Jung N, Cornely OA, Lehmann C, Tessarz P, Altmüller J, Nürnberg P, Kashkar H, Klein F, Koch M, Rybniker J. Long-lived macrophage reprogramming drives spike protein-mediated inflammasome activation in COVID-19. EMBO Mol Med. 2021 Aug 9;13(8):e14150. doi: 10.15252/emmm.202114150.

Cramer P, Tausch E, von Tresckow J, Giza A, Robrecht S, Schneider C, Fürstenau M, Langerbeins P, Al-Sawaf O, Pelzer BW, Fink AM, Fischer K, Wendtner CM, Eichhorst B, Kneba M, Stilgenbauer S, Hallek M. Durable remissions following combined targeted therapy in patients with CLL harboring TP53 deletions and/or mutations. Blood. 2021 Nov 11;138(19):1805-1816. doi: 10.1182/blood.2020010484.

Pan H, Renaud L, Chaligne R, Bloehdorn J, Tausch E, Mertens D, Fink AM, Fischer K, Zhang C, Betel D, Gnirke A, Imielinski M, Moreaux J, Hallek M, Meissner A, Stilgenbauer S, Wu CJ, Elemento O, Landau DA. Discovery of Candidate DNA Methylation Cancer Driver Genes. Cancer Discov. 2021 Sep;11(9):2266-2281. doi: 10.1158/2159-8290.CD-20-1334.

Beckmann L, Berg V, Dickhut C, Sun C, Merkel O, Bloehdorn J, Robrecht S, Seifert M, da Palma Guerreiro A, Claasen J, Loroch S, Oliverio M, Underbayev C, Vaughn L, Thomalla D, Hülsemann MF, Tausch E, Fischer K, Fink AM, Eichhorst B, Sickmann A, Wendtner CM, Stilgenbauer S, Hallek M, Wiestner A, Zahedi RP, Frenzel LP. MARCKS affects cell motility and response to BTK inhibitors in CLL. Blood. 2021 Aug 19;138(7):544-556. doi: 10.1182/blood.2020009165.

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.

Selected Publications 2020

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.