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.

The most important publications of the Künkele Lab

For a complete publication list please click here.

Andersch L, Radke J, Klaus A, Schwiebert S, Winkler A, Schumann E, Grunewald L, Zirngibl F, Flemmig C, Jensen MC, Rossig C, Joussen A, Henssen A, Eggert A, Schulte JH, Künkele A. CD171- and GD2-specific CAR-T cells potently target retinoblastoma cells in preclinical in vitro testing. BMC Cancer. 2019 Sep9; 19(1):895. doi: 10.1186/s12885-019-6131-1. PubMed PMID: 31500597; PubMed

Künkele A, Brown C, Beebe A, Mgebroff S, Johnson AJ, Taraseviciute A, Rolczynski LS, Chang CA, Finney OC, Park JR, Jensen MC. Manufacture of Chimeric Antigen Receptor T Cells from Mobilized Cyropreserved Peripheral Blood Stem Cell Units Depends on Monocyte Depletion. Biol Blood Marrow Transplant. 2019 Feb;25(2):223-232. doi: 10.1016/j.bbmt.2018.10.004. Epub 2018 Oct 10. PubMed

Gottlieb A, Althoff K, Grunewald L, Thor T, Odersky A, Schulte M, Deubzer HE, Heukamp L, Eggert A, Schramm A, Schulte JH, Künkele A. RITA displays anti-tumor activity in medulloblastomas independent of TP53 status. Oncotarget. 2017 Apr 25;8(17):27882-27891. doi: 10.18632/oncotarget.15840. 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. PubMed

Künkele A, Wilm J, Holdt M, Lohmann D, Bornfeld N, Eggert A, Temming P, Schulte JH. Neoadjuvant/adjuvant treatment of high-risk retinoblastoma: a report from the German Retinoblastoma Referral Centre. Br J Ophthalmol. 2015 Jul;99(7):949-53. doi: 10.1136/bjophthalmol-2014-306222. Epub 2015 Jan 12. PubMed

Künkele A, Johnson AJ, Rolczynski LS, Chang CA, Hoglund V, Kelly-Spratt KS, Jensen MC. Functional Tuning of CARs Reveals Signaling Threshold above Which CD8+ CTL Antitumor Potency Is Attenuated due to Cell Fas-FasL-Dependent AICD. Cancer Immunol Res. 2015 Apr;3(4):368-79. doi: 10.1158/2326-6066.CIR-14-0200. Epub 2015 Jan 9. PubMed

Pajtler KW, Weingarten C, Thor T, Künkele A, Heukamp LC, Büttner R, Suzuki T, Miyata N, Grotzer M, Rieb A, Sprüssel A, Eggert A, Schramm A, Schulte JH. The KDM1A histone demethylase is a promising new target for the epigenetic therapy of medulloblastoma. Acta Neuropathol Commun. 2013 May 29;1:19. doi: 10.1186/2051-5960-1-19. PubMed

Künkele A, Engelhard M, Hauffa BP, Mellies U, Müntjes C, Hüer C, Eggert A, Schulte JH, Kremens B. Long-term follow-up of pediatric patients receiving total body irradiation before hematopoietic stem cell transplantation and post-transplant survival of >2 years. Pediatr Blood Cancer. 2013 Nov;60(11):1792-7. doi: 10.1002/pbc.24702. Epub 2013 Jul 24. PubMed

Künkele A, Jurklies C, Wieland R, Lohmann D, Bornfeld N, Eggert A, Schulte JH. Chemoreduction improves eye retention in patients with retinoblastoma: a report from the German Retinoblastoma Reference Centre. Br J Ophthalmol. 2013 Oct;97(10):1277-83. doi: 10.1136/bjophthalmol-2013-303452. Epub 2013 Jul 17. PubMed

Heukamp LC, Thor T, Schramm A, De Preter K, Kumps C, De Wilde B, Odersky A, Peifer M, Lindner S, Spruessel A, Pattyn F, Mestdagh P, Menten B, Kuhfittig-Kulle S, Künkele A, König K, Meder L, Chatterjee S, Ullrich RT, Schulte S, Vandesompele J, Speleman F, Büttner R, Eggert A, Schulte JH. Targeted expression of mutated ALK induces neuroblastoma in transgenic mice. Sci Transl Med. 2012 Jul 4;4(141):141ra91. doi: 10.1126/scitranslmed.3003967. PubMed

Künkele A, De Preter K, Heukamp L, Thor T, Pajtler KW, Hartmann W, Mittelbronn M, Grotzer MA, Deubzer HE, Speleman F, Schramm A, Eggert A, Schulte JH. Pharmacological activation of the p53 pathway by nutlin-3 exerts anti-tumoral effects in medulloblastomas. Neuro Oncol. 2012 Jul;14(7):859-69. doi: 10.1093/neuonc/nos115. Epub 2012 May 16. PubMed

Current methods established in the Künkele Lab

  • Cell culture: T cells, modified T cells expressing chimeric antigen receptors (CAR), neuroblastoma cell lines, medulloblastoma cell lines, retinoblastoma cell lines, organoids
  • Animal models: Patient-derived xenografts in mice (PDXs), cell line-derived xenografts in mice, spontaneous neuroblastoma model in genetically modified mice, human and mice CAR-T cells
  • 3D tumor models
  • Stable inducible expression systems
  • CRISPR/Cas9 technology
  • Transfections: transient/stable, lentiviral, retroviral, siRNAs/shRNAs/DNA-vectors
  • Gene expression analyses: qRT-PCR, qPCR
  • Cloning technologies
  • Luciferase-based reporter gene assays
  • Fluorescence activated cell sorting (FACS)
  • Magnetic activated cell sorting (MACS)
  • Protein analyses: western blotting, (co-)immunoprecipitation
  • Cytokine analyses with ELISA (Enzyme-linked Immunosorbent Assay)
  • Proliferation-, apoptosis-, differentiation- and colony formation assays

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.
  • Retinoblastoma: Univ.-Prof. Dr.med. Antonia Joussen, Medical Director Department of Ophthalmology, Charité-Universitätsmedizin Berlin.
  • 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
  • Development of tumor models: Cellbricks GmbH, EPO - Experimental Pharmacology and Oncology Berlin-Buch GmbH, CPO cellular phenomics and oncology Berlin-Buch GmbH
  • Neuropathology: Dr. Josefine Radke, Department of Neuropathology, CharitéUniversitätsmedizin Berlin