Tumorimmunologie

Adoptive T cell therapy using genetically engineered T cells expressing a chimeric antigen receptor (CAR) has revolutionized the treatment modalities in leukemia and lymphoma for more than a decade now. However, the translation of the successes to solid tumor diseases is rather slow. Within our team, we are dedicated to the development and manufacturing of novel and safe CARs that can be applied in the clinics. For instance, we have developed two novel CAR-T cell products for the use in breast cancer patients that target the cancer testis antigen NY-BR-1 and prevent cross-reactivity against healthy brain tissue. Our team analyzed both for efficiency and safety in vitro and in vivo in a xenotransplant and target transgenic mouse model.

In close collaboration with Dr. Richard Harbottle (DKFZ, DNA Vector Lab), we have developed a novel DNA-plasmid-based vector system for the GMP-grade manufacturing of CAR‑T cells that makes the use of (lenti)viral vectors dispensable. These vectors, which we named nS/MARt, do not possess the risk of genotoxicity but confer the same characteristics as viral vectors, e.g. stable expression (Bozza M et al, 2021, SciAdv, 7(16):eabf1333). As desired side effects, this novel manufacturing method will reduce overall costs for CAR-T cell production and allows the establishment of a de-centralized manufacturing process.

As an accompanying project, we are developing novel, specific CAR inhibitors that can temporarily impair their function and therefore maybe useful to lower clinical side effects. These inhibitors are based on non-pathological AAV-particles that present specific CAR-epitopes on their surface. Currently, we are validating our patented technology (WO/2019/043081) by identification of matching pairs of CARs and CARAAVs.

In a joint effort together with the CCU “Applied Tumor Immunity” (DKFZ) we try to accelerate the identification of de novo, tumor antigen-specific CAR constructs to be used in personalized cancer immunotherapy. To achieve this goal, a unique cellular CAR library screening platform is generated by taking advantage of novel nanoSMAR vectors and the Lightning System, a single-cell microfluidic system from Berkeley Lights.

Contact: Patrick Schmidt, PhD - patrick.schmidt@nct-heidelberg.de

We aim for the development and routine application of personalized TCR gene-engineered adoptive T cell therapies (ACT) targeting individual- as well as shared patient-derived neoepitopes. Based on our implemented state-of-the-art tumor mutagenome analysis as well as in silico T cell epitope prediction pipeline that is combined with our highly sensitive peptide-loaded MHC-multimer-guided antigen-specific T cell discovery workflow (CCU “Applied Tumor Immunity” (DKFZ), Antigen Presentation and T/NK Cell Activation, PD Dr. Momburg), we have already identified and validated several neoepitope-specific TCRs across various solid tumor identities. This TCR discovery workflow is further augmented by our own robust pMHC in-house production platform that outperforms commercially available reagents and allows us to provide complete coverage of the individual patients’ HLA allotypes. By incorporating DNA-barcoded pMHC-I multimer reagents into the workflow, we link multiplexed tumor antigen recognition with their associated TCR repertoire and T cell differentiation phenotype at single cell resolution, which further improves the selection of TCR / antigen pairs for validation prior to their usage in an ACT.

The PIKT project is a pilot project funded by the Dietmar Hopp foundation for the development of personalized cellular immunotherapies for solid tumor diseases. The scope of the project is the identification and cloning of individual neoepitope specific TCRs and the manufacturing of clinical-grade transgenic TCR-T cell products as a proof of concept. For TCR discovery, the above pipeline is applied. We have screened more than 35 cases and so far and T cell receptors for nine cases were successfully identified, cloned and validated.

To deepen our knowledge about T cell (dys)function, a panel of analytical methods to determine T cell fitness of patients before and after T cell therapy by multiparametric flow cytometry has been established in close collaboration with Dr. Paula Roberti, CCU “Applied Tumor Immunity” (DKFZ). In an individual case, the association between a high T cell fitness signature in the infusion product and long-term persistence of TCR-T cells in the patient with sustained reactivity to the tumor target ex vivo could be shown. In addition to T cell fitness, other tumor-intrinsic resistance mechanisms are explored with the goal to not only find resistance mechanisms to cellular therapies, but also to develop new tools to overcome them.

Contact:

• Marten Meyer, PhD - marten.meyer@dkfz-heidelberg.de
• Maria Paula Roberti, PhD - mariapaula.roberti@nct-heidelberg.de
• Inka Zörnig, PhD - inka.zoernig@nct-heidelberg.de

We have established manufacturing protocols for the GMP production of CAR-T cells and transgenic TCR-T cells using the semi-automated manufacturing platform CliniMACS Prodigy from Miltenyi Biotec. Protocols are available for the lentiviral vector based production as well as for production using DNA-plasmid vectors. With strong support by the Dietmar Hopp Foundation, we envision to establish a TCR discovery lab and GMP manufacturing facility for cellular medical products in 2025. The so called “Center for Innovative Therapies” will harbor 320 sqm of clean room space classified in levels B, C and D in which production of individualized tumor vaccines and patient specific cell infusions will be possible. All rooms will be equipped with state-of-the-art devices in order to be able to perform any current manufacturing process possible. A dedicated and trained team will constantly establish SOPs and realize the regulatory prerequisites. This will enable us to develop innovative therapy approaches and produce study drugs for Phase I trials at the NCT, the UKHD, for partner sites within the NCT network as well as to serve as manufacturer for approved cellular drugs.

Contact: Patrick Schmidt, PhD - patrick.schmidt@nct-heidelberg.de

For the testing and individualized validation of CAR/TCR-T cell approaches, tissue co-cultures/organoids are applied with subsequent in depth analyses of the cell products, co-culture supernatants and tissue/organoids by multimarker FACS, IHC, virtual microscopy, and semi-automated image analyses.

Contact:  

• Patrick Schmidt, PhD - patrick.schmidt@nct-heidelberg.de
• Inka Zörnig, PhD - inka.zoernig@nct-heidelberg.de

The PROMISE trial (PRedictability of Outcome based on iMmunological sIgnatureS in lung cancEr – an explorative investigation) is a collaborative exploratory biomarker study with the Thoraxklinik, the UKHD and the DKFZ, funded by the Dietmar Hopp foundation. The trial was initiated in 2018 and was open for recruitment until summer 2022. Overall, 136 patients have been enrolled in the study.The trial focusses on the identification of predictive markers or signatures for response and resistance to current standard of care treatment in stage IV non-small cell lung cancer (NSCLC) patients. Biomaterials, such as tissue, serum, plasma, PBMCs, breath condensates and stool samples, are systematically collected longitudinally and will be analyzed by e.g. WGS/WES, epigenomics, PanelSeq, RNASeq, Nanostring, metagenomics, metabolomics, immunohistochemistry and virtual microscopy with semi-automated image analyses, multiplex Cytokine/Chemokine assays, mass spectrometry, RAMAN spectroscopy and flow cytometry for naïve/memory/effector immune cell phenotyping. Furthermore, we are analyzing the TCR repertoire in responders/non-responders in our clinical cohorts by deep sequencing of TCR variable beta (Vβ) (TCRB) chain. This will allow us to evaluate and measure the clonality and diversity of T cells that can arise in response to tumor-associated antigens and to investigate whether these changes could be predictive markers of response.

Due to the advancement in high-throughput technologies, we incorporate heterogeneous data from multiple technologies to improve our understanding of the molecular dynamics associated with biological processes. Pipelines for multi-omics data analysis comprising MHC I/II epitope prediction and V(D)J enrichment immune profiling analysis have been set up. Further, we are developing machine- and deep learning approaches on an in-house generated graph database enabling us to extract relevant biological interpretations and to identify novel relationships by integrating high-dimensional data. For an in-depth understanding of the “individual disease” and for the identification of predictive biomarkers, an integrated analysis of the multiomics (Charoentong P et al, 2017, Cell Rep, 18(1):248-62) and wet lab data is performed.

Contact:

• Pornpimol Charoentong, PhD - pornpimol.charoentong@nct-heidelberg.de
• Inka Zörnig, PhD - inka.zoernig@nct-heidelberg.de

The translational research program of the NEOMUN trial (“NEOadjuvant anti-PD-1 imMUNotherapy in resectable NSCLC: The NEOMUN Trial”) is supported by NCT POC trial funding and is carried out in the CCU Applied Tumor Immunity (DKFZ) /NCT TME Unit in collaboration with PD Dr. M. Eichhorn and colleagues (Surgery Department, Thoraxklinik Heidelberg). A major aim is the analysis of possible therapy-associated changes in different tumor regions (e.g. core vs. border), lymph nodes or healthy lung tissue and the potential identification of predictive markers.

Contact:

• Pornpimol Charoentong, PhD - pornpimol.charoentong@nct-heidelberg.de
• Inka Zörnig, PhD - inka.zoernig@nct-heidelberg.de

The IReP trial (“Immune Response Prediction” trial; funded by the Dietmar Hopp Foundation) is an exploratory study evaluating the potential of immune signature profiling for predicting response to neoadjuvant chemoimmunotherapy in patients with resectable stage II/III non-squamous NSCLC. An in depth analysis of the tumor microenvironment and genomic alterations is performed in pre- and post-treatment tissue and data is correlated with response as defined by Major pathologic response. The trial is performed in collaboration with PD Dr. M. Eichhorn (Department of Surgery, Thoraxklinik). 

Contact:

• Pornpimol Charoentong, PhD - pornpimol.charoentong@nct-heidelberg.de
• Inka Zörnig, PhD - inka.zoernig@nct-heidelberg.de

To bring Fecal Microbiota Transfer (FMT) therapy to the standard of care, the open challenge is to identify cancer patients at risk of gut-related therapy failure who are amenable to FMT. We are currently collaborating with clinicians and scientists at UKHD and NCT (Dr. C. Rauber) to conduct the "Mucosome" study (S-633/2021) with the aim of identifying gut biomarkers in patients with extraintestinal cancers that can be used for the selection of patients who will potentially benefit from FMT and to identify stool donors that can be used pharmacologically in a tailored donor-recipient approach in cancer therapy.

Contact: Maria Paula Roberti, PhD - mariapaula.roberti@nct-heidelberg.de

In multifocal hepatocellular carcinoma, the recently introduced first-line therapy Atezolizumab/Bevacizumab leads to an objective response rate of only 27%, which is superior to previous first-line therapies but highlights the urgent need for further therapeutic improvement. Together with Dr. C. Rauber (UKHD) and Dr. M. Dill (UKHD/DKFZ), who conduct "The FLORA trial: Fecal Microbiota Transfer in Liver Cancer to Overcome Resistance to Atezolizumab/Bevacizumab” Dr. M. Paula Roberti and the CCU team will perform the accompanying translational research program. Part of the “NCT Proof-of-Concept Trial Program” within the framework of Liver Cancer Center Heidelberg (LCCH), FLORA is a randomized, single-blind, placebo-controlled phase II clinical trial, which aims to assess immunogenicity and safety of FMT in combination with standard of care immunotherapy in advanced hepatocellular carcinoma.

Contact: Maria Paula Roberti, PhD - mariapaula.roberti@nct-heidelberg.de

The comprehensive analysis of tumor-host cell interactions, especially focusing on immune cells and the spatial analysis of the tumor microenvironment by immunohistochemistry (IHC), multiplex analyses, virtual microscopy and semi-automated image analysis is another main objective of the division. The molecular mechanisms underlying the clinical impact and the causes of the observed differences in the immunological tumor micro-milieu are not yet fully understood. Therefore, the dedicated analysis of the tumor compartments (stroma, epithelium, invasion front, and surrounding tissue) with respect to localization of immune cells and cytokine profiling in the light of the clinical course will yield a better image of the complex relations and eventually support the development of new drugs and targeted treatment approaches for patients. Together with the CCU Applied Tumor Immunity (DKFZ), the NCT TME Unit and the NCT Unit for „Applied Tumor Therapy and Combinational Strategies“, collaboration projects and clinical studies, especially biomarker trials such as PROMISE, NEOMUN and also programs such as CATCH, COGNITION and MASTER are supported. Further, the group is closely interacting with other units of the NCT Immunotherapy Program, which has been established to support drug- and target-development for novel immunotherapies (e.g. antibody-based strategies, cellular approaches and vaccines). This includes pre-clinical testing in cell culture, in human tissue model systems, organoids as well as small animal models and subsequent in-depth analysis of the tumor microenvironment.

Contact:

• Bénédicte Lenoir, PhD - benedicte.lenoir@nct-heidelberg.de
• Inka Zörnig, PhD - inka.zoernig@nct-heidelberg.de

• Warwas KM, Meyer M, Gonçalves M, Moldenhauer G, Bulbuc N, Knabe S, Luckner-Minden C, Ziegelmeier C, Heussel CP, Zörnig I, Jäger D, Momburg F. Co-Stimulatory Bispecific Antibodies Induce Enhanced T Cell Activation and Tumor Cell Killing in Breast Cancer Models. Front Immunol 12:719116 (2021). DOI: 10.3389/fimmu.2021.719116.
• Eichhorn F, Klotz LV, Kriegsmann M, Bischoff H, Schneider MA, Muley T, Kriegsmann K, Haberkorn U, Heussel CP, Savai R, Zoernig I, Jaeger D, Thomas M, Hoffmann H, Winter H, Eichhorn ME. Neoadjuvant anti-programmed death-1 immunotherapy by pembrolizumab in resectable non-small cell lung cancer: First clinical experience. Lung Cancer. 153:150-157 (2021). doi: 10.1016/j.lungcan.2021.01.018.
• Bozza M, De Roia A, Correia MP, Berger A, Tuch A, Schmidt A, Zörnig I, Jäger D, Schmidt P, Harbottle RP. A nonviral, nonintegrating DNA nanovector platform for the safe, rapid, and persistent manufacture of recombinant T cells. Sci Adv 7(16):eabf1333 (2021). DOI: 10.1126/sciadv.abf1333.
• Ran T, Eichmüller SB, Schmidt P, Schlander M. Cost of decentralized CAR T-cell production in an academic nonprofit setting. Int J Cancer. 147(12):3438-3445 (2020). DOI: 10.1002/ijc.33156.
• Rölle A, Meyer M, Calderazzo S, Jäger D, Momburg F. Distinct HLA-E Peptide Complexes Modify Antibody-Driven Effector Functions of Adaptive NK Cells. Cell Rep 24:1967-76.e4 (2018). DOI: 10.1016/j.celrep.2018.07.069.
• Michels J, Becker N, Suciu S, Kaiser I, Benner A, Kosaloglu-Yalcin Z, Agoussi S, Halama N, Pawlita M, Waterboer T, Eichmüller SB, Jäger D, Eggermont AMM, Zörnig I. Multiplex bead-based measurement of humoral immune responses against tumor-associated antigens in stage II melanoma patients of the EORTC18961 trial. Oncoimmunology. 12;7(6):e1428157 (2018). DOI: 10.1080/2162402X.2018.1428157.
• Halama N, Zoernig I, Berthel A, Kahlert C, Klupp F, Suarez-Carmona M, Suetterlin T, Brand K, Krauss J, Lasitschka F, Lerchl T, Luckner-Minden C, Ulrich A, Koch M, Weitz J, Schneider M, Buechler MW, Zitvogel L, Herrmann T, Benner A, Kunz C, Luecke S, Springfeld C, Grabe N, Falk CS, Jäger D. Tumoral immune cell exploitation in colorectal cancer liver metastases can be targeted effectively by anti-CCR5 therapy in cancer patients. Cancer Cell 29(4):587-601 (2016). DOI: 10.1016/j.ccell.2016.03.005.
• Gnjatic S, Ritter E, Büchler MW, Giese NA, Brors B, Frei C, Murray A, Halama N, Zörnig I, Chen YT, Andrews C, Ritter G, Old LJ, Odunsi K, Jäger D. Seromic profiling of ovarian and pancreatic cancer. Proc Natl Acad Sci U S A 107(11):5088-93 (2010). DOI: 10.1073/pnas.0914213107.
• Seil I, Frei C, Sültmann H, Knauer SK, Engels K, Jäger E, Zatloukal K, Pfreundschuh M, Knuth A, Tseng-Chen Y, Jungbluth AA, Stauber RH, Jäger D. The differentiation antigen NY-BR-1 is a potential target for antibody-based therapies in breast cancer. Int J Cancer. 120(12):2635-42 (2007). DOI: 10.1002/ijc.22620.
• Jäger D, Stockert E, Güre AO, Scanlan MJ, Karbach J, Jäger E, Knuth A, Old LJ, Chen YT. Identification of a tissue-specific putative transcription factor in breast tissue by serological screening of a breast cancer library. Cancer Res 61(5):2055-2061 (2001).

Group Head

Prof. Dr. Dirk Jäger

Head of 'Tumor Immunology' and Head of NCT Program 'Immunotherapy', NCT Heidelberg
Director of the Medical Oncology Department, Heidelberg University Medical Center (UKHD)
Director of the National Center for Tumor Diseases (NCT) Heidelberg
Head of the CCU Applied Tumor Immunity Unit and the Applied Tumor Immunity Group (DKFZ)

Secretariat Medical Oncology
Birgit Eberle
Tel: +49 6221 56-7229
Email: birgit.eberle@nct-heidelberg.de

Research Associates

Liam Bartels (DKFZ)
Position: Researcher
Area of expertise: Computational pathology, Machine learning, Medical sample imaging
Email: liam.bartels@dkfz-heidelberg.de

Eren Boga (DKFZ)
Position: PhD Student
Title: „Generation of a cellular CAR library for rapid identification of antigen-specific, functional CAR constructs"
Email: eren.boga@nct-heidelberg.de

Daniel Browne
Position: IT System Administrator
Email: daniel.browne@bioquant.uni-heidelberg.de

Pornpimol Charoentong, PhD
Position: Senior Research Associate, Bioinformatician
Area of expertise: Genomics and Bioinformatics, Computational System Biology, Biostatistics, Management of biological databases
Email: Pornpimol.Charoentong@nct-heidelberg.de

Sarah Glenz (DKFZ)
Position: PhD Student
Title: "Structural and functional consequences of AAV capsid engineering"
Email: sarah.glenz@nct-heidelberg.de

Niels Grabe, PhD (Prof. Dr.)
Position:  Professor at University Medicine Göttingen and NCT & University Hospital Heidelberg, Scientific Director of TIGA Center for Medical Informatics / Bioinformatics / AI, Heidelberg
Area of expertise: AI based digital pathology, image analysis, medical informatics and bioinformatics
Email:  niels.grabe@bioquant.uni-heidelberg.de

Bénédicte Lenoir, PhD (DKFZ)
Position: Senior Research Associate, project coordinator "NCT TME Unit"
Area of expertise: Immunohistology, image analysis, project management
Email: benedicte.lenoir@nct-heidelberg.de

Yanhong Lyu, PhD (DKFZ)
Position: Postdoc, Bioinformation
Area of expertise: Bioinformatics, Genomics
Email: y.lyu@dkfz-heidelberg.de

Marten Meyer, PhD (UKHD/DKFZ)
Position: Senior Research Associate
Area of expertise: translational research, TCR discovery, recombinant protein production
Email: marten.meyer@dkfz-heidelberg.de

Frank Momburg, PhD (DKFZ/UKHD)
Position: Senior Research Associate, Group Leader DKFZ
Area of expertise: translational research, TCR discovery, recombinant protein production
Email: frank.momburg@dkfz-heidelberg.de

Julia Pollmann, PhD
Position: Senior Research Associate
Area of expertise: Innate Immunity, Immuno-oncology, chronic viral infections
Email: j.pollmann@dkfz-heidelberg.de 

Ferdinand Popp (DKFZ)
Position: Research Associate
Area of expertise: Cancer and Molecular Biology, Multi-omics and clinical data analysis, Machine learning & Data Science
Email: f.popp@dkfz-heidelberg.de

Maria Paula Roberti, PhD (DKFZ)
Position: Senior Research Associate, Deputy Head of the CCU Applied Tumor Immunity (DKFZ)
Area of expertise: Immunotherapy, Translational Research, Gut microbiota and immunity
Email: mariapaula.roberti@nct-heidelberg.de

Alexander Rölle, PhD
Position: Senior Research Associate
Area of expertise: Innate Immunity, Cellular Immunology, Tumor Microenvironment
Email: a.roelle@nct-heidelberg.de

Patrick Schmidt, PhD (UKHD/DKFZ)
Position: Senior Research Associate, Team leader "Cellular therapies"
Area of expertise: Cellular therapy, CAR-T cells, TCR-T cells
Email: patrick.schmidt@nct-heidelberg.de

Silke Uhrig-Schmidt, PhD (DKFZ)
Position: Senior Research Associate
Area of expertise: AAV capsid design, cancer vaccination, gene and cell therapy
Email: silke.uhrig-schmidt@nct-heidelberg.de

Nektarios Valous, PhD (DKFZ)
Position: Senior Research Associate
Area of expertise: Computational image processing, computational biomedicine, interdisciplinary physics
Email: nek.valous@nct-heidelberg.de

Katharina von Werthern (DKFZ)
Position: Master Student

Heiko Weyd, PhD (DKFZ)
Position: Senior Research Associate
Area of expertise: Immunology, Antibodies, Apoptosis, T cells, Tolerance
Email: h.weyd@dkfz-heidelberg.de

Inka Zörnig, PhD
Position: Senior Research Associate, Project coordination and lab head
Area of expertise: translational research, biomarker discovery, immunotherapy
Email: inka.zoernig@nct-heidelberg.de 

Technical Assistance

Selina Börsig (DKFZ/UKHD)

Martina Busacker-Scharpff

Veronika Geißler

Isabella Gosch

Petra Hill

Iris Kaiser

Annette Köster

Alexandra Krauthoff (DKFZ)

Claudia Luckner-Minden

Claudia Tessmer (DKFZ)

Alexandra Tuch (DKFZ)

Nina Wilhelm (DKFZ)

Claudia Ziegelmeier

Former Members

Dr. Aileen Berger

Dr. Julia Bitzer

Dr. Lasse Neukirch

Dr. Rodrigo Moraleda

Dr. Zeynep Koşaloğlu-Yalçın