The Künkele Lab
CAR-T cell therapy as new treatment approach for neuroblastoma
More than half of high-risk patients with neuroblastoma, a common childhood tumor, currently die of disease.
To better attack this cancer in children, we are developing new immunotherapies using T cells, a type of white blood cell, removed from patient blood and modified in the lab.
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Research of the Künkele Lab
T cells can destroy virus-infected cells and tumor cells. By changing proteins on their cell surface, it is possible to "teach" them to find neuroblastoma cells in the body and to arm them with so-called chimeric antigen receptors (CARs) to kill the tumor cells. The armed CAR-T cells are reintroduced into the patient as a new weapon to fight the cancer.
CAR-T cell therapy has worked extremely well against leukemia, but not yet against solid tumors, as these can better protect themselves from the immunotherapy attack. Our research seeks to identify and reduce these defense mechanisms, which are currently preventing cure. Together with national and international cooperation partners, we will investigate the influence of tumor cells, their environment and surrounding blood vessels on CAR-T cells to find out how these factors allow tumor cells to escape immunotherapy, then use this new knowledge to find efficient ways to shut off the devastating tumor defense mechanisms.
News
The 10 important publications of the Künkele Lab
For a complete publication list please click here.
Sulejmani O, Grunewald L, Andersch L, Schwiebert S, Klaus A, Winkler A, Astrahantseff K, Eggert A, Henssen AG, Schulte JH, Anders K, Künkele A. Inhibiting Lysine Demethylase 1A Improves L1CAM-Specific CAR T Cell Therapy by Unleashing Antigen-Independent Killing via the FAS-FASL Axis. Cancers (Basel). 2021 Oct 31;13(21):5489. doi: 10.3390/cancers13215489. PMID: 34771652; PubMed
Zirngibl F, Ivasko SM, Grunewald L, Klaus A, Schwiebert S, Ruf P, Lindhofer H, Astrahantseff K, Andersch L, Schulte JH, Lode HN, Eggert A, Anders K, Hundsdoerfer P, Künkele A. GD2-directed bispecific trifunctional antibody outperforms dinutuximab beta in a murine model for aggressive metastasized neuroblastoma. J Immunother Cancer. 2021 Jul;9(7):e002923. doi: 10.1136/jitc-2021-002923. PMID: 34285106; PMCID: PMC8292814. PubMed
Grunewald L, Lam T, Andersch L, Klaus A, Schwiebert S, Winkler A, Gauert A, Heeren-Hagemann AI, Astrahantseff K, Klironomos F, Thomas A, Deubzer HE, Henssen AG, Eggert A, Schulte JH, Anders K, Kloke L, Künkele A. A Reproducible Bioprinted 3D Tumor Model Serves as a Preselection Tool for CAR T Cell Therapy Optimization. Front Immunol. 2021 Jun 29;12:689697. doi: 10.3389/fimmu.2021.689697. PMID: 34267756; PMCID: PMC8276678 PubMed
Vitanza NA, Johnson AJ, Wilson AL, Brown C, Yokoyama JK, Künkele A, Chang CA, Rawlings-Rhea S, Huang W, Seidel K, Albert CM, Pinto N, Gust J, Finn LS, Ojemann JG, Wright J, Orentas RJ, Baldwin M, Gardner RA, Jensen MC, Park JR. Locoregional infusion of HER2-specific CAR T cells in children and young adults with recurrent or refractory CNS tumors: an interim analysis. Nat Med. 2021 Jul 12. doi: 10.1038/s41591-021-01404-8. Epub ahead of print. PMID: 34253928. PubMed
Textor A, Grunewald L, Anders K, Klaus A, Schwiebert S, Winkler A, Stecklum M, Henssen AG, Höpken UE, Eggert A, Schulte JH, Jensen MC, Blankenstein T, Künkele A. CD28 Co-Stimulus Achieves Superior CAR T Cell Effector Function against Solid Tumors than 4-1BB Co-Stimulus, PubMed
Ali S, Toews K, Schwiebert S, Klaus A, Winkler A, Grunewald L, Oevermann L, Deubzer HE, Tüns A, Jensen MC, Henssen AG, Eggert A, Schulte JH, Schwich E, Rebmann V, Schramm A, Künkele A. Tumor-Derived Extracellular Vesicles Impair CD171-Specific CD4+ CAR T Cell Efficacy. Front Immunol. 2020 Mar31;11:531. doi: 10.3389/fimmu.2020.00531. PMID: 32296437; PMCID: PMC7137471. PubMed
Koche RP, Rodriguez-Fos E, Helmsauer K, Burkert M, MacArthur IC, Maag J, Chamorro R, Munoz-Perez N, Puiggròs M, Dorado Garcia H, Bei Y, Röefzaad C, Bardinet V, Szymansky A, Winkler A, Thole T, Timme N, Kasack K, Fuchs S, Klironomos F, Thiessen N, Blanc E, Schmelz K, Künkele A, Hundsdörfer P, Rosswog C, Theissen J, Beule D, Deubzer H, Sauer S, Toedling J, Fischer M, Hertwig F, Schwarz RF, Eggert A, Torrents D, Schulte JH, Henssen AG. Extrachromosomal circular DNA drives oncogenic genome remodeling in neuroblastoma. Nat Genet. 2020 Jan;52(1):29-34. doi: 0.1038/s41588-019-0547-z. Epub 2019 Dec 16. Erratum in: Nat Genet. 2020 Feb 27;: PMID: 31844324; PubMed
Künkele A, Taraseviciute A, Finn LS, Johnson AJ, Berger C, Finney O, Chang CA, Rolczynski LS, Brown C, Mgebroff S, Berger M, Park JR, Jensen MC. Preclinical Assessment of CD171-Directed CAR T-cell Adoptive Therapy for Childhood Neuroblastoma: CE7 Epitope Target Safety and Product Manufacturing Feasibility. Clin Cancer Res. 2017 Jan 15;23(2):466-477. doi: 10.1158/1078-0432.CCR-16-0354. Epub 2016 Jul 7. PMID: 27390347. PubMed
Hölting TLB, Cidre-Aranaz F, Matzek D, Popper B, Jacobi SJ, Funk CM, Geyer FH, Li J, Piseddu I, Cadilha BL, Ledderose S, Zwilling J, Ohmura S, Anz D, Künkele A, Klauschen F, Grünewald TGP, Knott MML. Neomorphic DNA-binding enables tumor-specific therapeutic gene expression in fusion-addicted childhood sarcoma. Mol Cancer. 2022 Oct 13;21(1):199. doi: 10.1186/s12943-022-01641-6. PMID: 36229873; PubMed
Schmelz K, Toedling J, Huska M, Cwikla MC, Kruetzfeldt LM, Proba J, Ambros PF, Ambros IM, Boral S, Lodrini M, Chen CY, Burkert M, Guergen D, Szymansky A, Astrahantseff K, Kuenkele A, Haase K, Fischer M, Deubzer HE, Hertwig F, Hundsdoerfer P, Henssen AG, Schwarz RF, Schulte JH, Eggert A. Spatial and temporal intratumour heterogeneity has potential consequences for single biopsy-based neuroblastoma treatment decisions. Nat Commun. 2021 Nov 23;12(1):6804. doi: 10.1038/s41467-021-26870-z. PMID: 34815394; PubMed
Current methods established in the Künkele Lab
- Cell culture: primary T cell culture, large spectrum of neuroblastoma, medulloblastoma and retinoblastoma cell lines
- Production of CAR-T cells using lentiviral and retroviral gene transfers as well as virus-free CAR-T cell production using CRISPR/Cas9 knockin
- 3D tumor models (organoids, 3D-bioprint)
- CRISPR knockout screening
- Live cell Imaging (Incucyte)
- Transwell-Migration Assays
- Digital droplet PCR
- Immunhistology (Kryo/FFPE)
- Luciferase-based killing assay
- Flow cytometry (FACS with up to 15 colors, Fortessa X-20) and cell sorting (Aria III)
- Magnetic cell sorting (MACS)
- Preclinical animal models (PDX/ xenograft/ syngeneic tumors)
- Adoptive T cell therapy with human and murine CAR T cells
- Gene expression analysis: RT-PCR
Selected cooperation partners
Neuroblastoma: Prof. Dr. J. H. Schulte, PD Dr. H. E. Deubzer and Dr. med. Anton G. Henssen, all Department of Pediatrics, Division of Oncology and Hematology, Charité-Universitätsmedizin Berlin; PD Dr. A. Schramm, Department of pediatric oncology, University Duisburg-Essen; Dr. Jan Molenaar, Center for Pediatric Oncology, Princess maxima center, Utrecht; Prof. Dr. Martin Eilers und Prof. Dr. Gabriele Büchel, Universität Würzburg;
Retinoblastoma: Univ.-Prof. Dr.med. Antonia Joussen, Medical Director Department of Ophthalmology, Charité-Universitätsmedizin Berlin; Prof. Dr. Ulrich Schraermeyer, Universitätsklinikum Tübingen
CAR- and TCR-T cell therapy: Prof. Dr. T. Blankenstein and Prof. Dr. G. Willimsky, both at the MDC Berlin-Buch; Prof Dr. C. Rössig, Department of Pediatrics, University Hospital Münster; Prof. Dr. L. Uharek, Charité-Universitätsmedizin Berlin; Miltenyi company in Bergisch-Gladbach; Dr. M. Hudeczek, Department of internal medicine, University Hospital Würzburg; Prof. Dr. J. Park and Prof. Dr. M. C. Jensen, both Seattle Children's Research Institute.
CAR-NK cell therapy: Prof. Dr. L. Uharek and PD Dr. O. Pennack, both at the Charité-Universitätsmedizin Berlin.
Immunology: PD Dr. S. Kobold, LMU Munich; Prof. Dr. K. S. Lang, Department of Immunology University Duisburg-Essen; PD Dr. rer. Medic. Vera Rebmann, Transfusion Medicine, Universitätsklinikum Essen; Dr. med. Judith Feucht, Universitätsklinikum Tübingen; PD Dr. Uta Höpken, MDC Berlin-Buch;
Development of tumor models: Cellbricks GmbH, EPO - Experimental Pharmacology and Oncology Berlin-Buch GmbH
Pathology: Prof. Dr. med. David Horst, Charité Universitätsmedizin Berlin
scSeq: Prof. Dr. Nikolaus Rajewsky, BIMSB; MDC Berlin-Buch;