DNA repair, hypoxia and genomic instability in metastatic prostate cancer (DRAGOON): Mechanistic insights and new treatment options

Dissemination of tumour cells and metastatic spread as well as cancer resistance to standard treatment are still the common cause of treatment failure and eventually death in prostate cancer patients. A better understanding the biology of the above-mentioned features will indeed lead to more effective cancer therapy options in prostate cancer. To that end, molecular techniques mixed with scientific experience of the medical scientist Dr. Mansour complemented by clinical medical oncology expertise of Dr. Oing are gathered to promote the current bidirectional translational research project entitled DRAGOON. In DRAGOON, several high throughput techniques in various in vivo, in vitro and ex vivo models will be employed to address features of dissemination and metastasis as well as endocrine resistance in this context, by examining incurable prostate cancer at

(i) an early (hormone-sensitive, node-positive) (subproject 1) and also (ii) a late (metastatic castration-resistant) (subproject 2) stage of metastatic disease as a special exemplar, taking into account the complexity of advanced prostate cancer. Both subprojects aim to identify distinct signatures within the genome, transcriptome, epigenome and/or kinome, which contribute to prostate cancer dissemination, metastasis and progression to castration resistant phenotype, which will assist in developing new biomarkers and novel, biomarker-driven, personalised therapeutic strategies (subproject 3).

Contact and affiliation:

PD Dr. rer. nat Wael Y. Mansour

Laboratory for Radiobiology & Experimental Radiation Oncology
w.mansour@uke.de
+49 (0) 40 7410 56520
Website: Regulation of DNA Double Strand Break Repair in Tumors

Dr. med. Christoph Oing

II. Medical Clinic and Polyclinic for Oncology and Hematology
c.oing@uke.de
+49 (0) 40 7410 55139

NUAcT Fellow
NU Translational & Clinical Research Institute
CRUK NU Drug Discovery Unit
Newcastle University

Honorary Consultant Medical Oncologist Early Clinical Trials Unit
Sir Bobby Robson Cancer Trials Research Centre
Northern Centre for Cancer Care@Freeman Hospital
Newcastle Hospitals NHS Foundation Trust

Dissecting the metastatic cascade to improve the clinical management of ovarian cancer

Phenotypically and genetically heterogeneous tumors are more aggressive, have a higher relapse rate, and are more resilient to therapy. Metastatic outgrowth of disseminated tumor cells is thought to be fueled by clonal cooperation of tumor clones from early and late stages of cancer evolution. Circulating tumor cells (CTCs) can be detected in the peripheral blood and peritoneal liquid in most of the ovarian cancer patients in the different stages of the disease. Literature suggests that CTCs that found their way into the blood stream may in general not be able to extravasate and colonialize outside the peritoneal cavity, but may in fact return to the primary site to perform so called self‐seeding. The process of growing the tumor from the “utside in”maybe one of the mechanisms of maintaining tumor heterogeneity and has significant clinical implications. This study will elucidate the metastatic cascade of ovarian cancer by investigating the natural cancer evolution through the analysis of single tumor cells on genomic, transcriptomic, proteomic, and methylomic (multi‐omic) level. Dissecting the metastatic cascade in ovarian cancer and understanding the evolutionary pathways of how and when metastasis takes place will greatly help in the management of cancer patients.

Contact and affiliation:

PD Dr. med. Katharina Prieske

Department of Gynecology
k.prieske@uke.de
+49 (0) 40 7410 54355

Dysplasiezentrum am Krankenhaus Jerusalem
Moorkamp 2-6
20357 Hamburg
+49 (0) 40 44190 450
Website

PD Dr. Simon A. Joosse, PhD

Department of Tumor Biology
s.joosse@uke.de
+49 (0) 40 741051970
Website

Histological, molecular and clinical characterization of MYB/MYBL1-altered gliomas

Currently, only few data on gliomas with alterations of the MYB or MYBL1 genes are available. Thus, they are often misdiagnosed, the prognosis is unclear and targeted therapies do not exist. I will assemble a patient cohort of approximately 100 MYB/MYBL1-altered gliomas to define subgroups with cluster analyses of DNA methylation data. Next, I will analyse the histology and immunohistochemical profile of these gliomas to investigate whether the histology, and also MRI features, correlate with the molecular subgroups and differ from other glioma entities. These data will also give insight whether MYB/MYBL1-altered gliomas, especially those with a high proliferation, may show a wider dissemination and metastasize. RNA sequencing will identify gene fusions in these gliomas, reveal differential gene expression compared to other gliomas and unravel signalling pathways active in MYB/MYBL1-altered gliomas. I will then express the gene fusions detected in murine cell lines in vitro to study the individual effect on cell growth. All these data will lead to a better understanding of the tumor biology of MYB/MYBL1-altered gliomas, improve diagnostic pipelines while also integrating molecular data, and indicate therapeutic targets for clinical use. Finally, I will analyse clinical data of patients with MYB/MYBL1-altered gliomas to get insight into the prognosis.

Contact and affiliation:

Dr. med. Annika Wefers

Institute of Neuropathology
Phone: 0152-22827602
Email: a.wefers@uke.de

Evaluation of innovative antibody fragment based targeted constructs as first line or combination treatment option for medulloblastoma and rhabdomyosarcoma using an in vitro cell-based Blood-Brain-Barrier Technology Platform

Partner: Dr. Ole Pless

The paediatric solid tumours medulloblastoma (MB, brain tumour) and rhabdomyosarcoma (RMS, soft tissue tumour) show overexpression of the epidermal growth-factor receptor (EGFR) and mutations or abnormal signalling in the PI3-kinase-pathway, which can lead to disseminated tumour growth. Both cancers metastasize; in RMS brain metastasis can develop. Conventional radio-and chemotherapy often results in severe long-term side-effects in young patients, such as problems with growth, learning or brain development. Therefore, targeted treatment options are urgently needed. We previously developed targeted therapies using antibody fragments (scFvs), targeting the EGFR. EGFR-specific scFvs as part of immunoconjugates, namely bispecific T-cell engagers (BiTEs) and immunotoxins (ITs), kill selectively tumour cells without affecting healthy cells. Using them in MB or RMS patients with brain metastasis, they must cross the Blood-Brain-Barrier (BBB), a protective layer in the brain. To evaluate the diffusion kinetics of the components, we want to test them in an innovative in vitro-BBB-model obtained from the cooperation partner. EGFR overexpression and constitutive PI3-kinase signalling are connected with maintenance of cancer stem-cells. Consequently, both EGFR-immunoconjugates should be used in combination with a well-known PI3-kinase inhibitor with established BBB penetrance. Having functional scFv-immunotherapeutics available, alternative cancer stem-cell related target proteins can be tested by exchanging the scFvs.

Contact and affiliation:

Dr. rer. nat. Judith Niesen
Research Institute Children’s Cancer Center Hamburg and Department of Pediatric Hematology and Oncology
j.niesen@uke.de
+49 (0) 40 426051214

Dr. rer. nat. Ole Pless
Institute for Molecular Biology and Applied Ecology at Fraunhofer IME Screening Port Hamburg
ole.pless@ime.fraunhofer.de

An integrated approach for drug target validation

Partner: Prof. Dr. Matthias Wilmanns

Distant metastasis is the major cause of cancer related death. Low RAI2 gene expression was initially identified at ITB/UCCH to be correlated with poor patient survival and in particular with the detection of disseminated tumor cells in the bone marrow of patients that were otherwise free of overt metastases. So far, the RAI2 protein function has not been annotated; but joint efforts of the applicant and the research team at EMBL funded by DFG have already shown that this protein acts as transcriptional co-regulator and that its inactivation in breast cancer cells directly leads and to dedifferentiation and deregulation of the estrogen response. This resulted in a high impact publication in Cancer Discovery (impact factor: 26.4). More recently both research teams showed that RAI2 counteracts oncogenic CtBP activity. Because active CtBPs suppress epithelial differentiation and propel cancer progression, we hypothesize that RAI2 mediated negative CtBP regulation might be exploited to treat cancer patients more efficiently. The major aim of the proposed project is to validate the potential of RAI2-mediated CtBP inactivation as new therapeutic concept using their integrated drug target validation approach and unique infrastructure at EMBL which will then become also available to other members of the MSNZ/UCCH consortium.

Contact and affiliation:

Dr. rer. nat. Stefan Werner
Department of Tumor Biology
st.werner@uke.de
+49 (0) 40 7410 57947

Prof. Dr. Matthias Wilmanns, PhD
Head of EMBL (European Molecular Biology Laboratory) Hamburg
EMBL Wilmanns Group

Promotion of bone-seeking tumors through the mechanosensory network in bone

Partner: Dr. Anton Davydok

Bone is a highly dynamic tissue undergoing a constant matrix remodeling activity regulated by mechanical stimuli. When breast and prostate cancers metastasize to bone, a favored site, the fine-tuned bone remodeling can hijacked while fragility fractures increase. Yet, the crucial role of the mechano-regulated bone turnover is still to be determined in the context of bone metastases. In this project we aim to assess the role of the mechanosensory lacuno-canalicular network of osteocytes on tumor progression with bone metastasis. We have designed an interdisciplinary project utilizing high-resolution imaging techniques, including synchrotron radiation-based small- and wide- angle X-ray scattering (SAXS/WAXS), to analyze the local processes whereby impaired local bone quality contributes to tumor progression.

Contact and affiliation:

Dr. rer. nat. Katharina Jähn-Rickert
Heisenberg Research Group Department of Osteology and Biomechanics
University Medical Center Hamburg-Eppendorf
Lottestr. 55a 22529
Hamburg Germany
Mail: k.jaehn@uke.de
Phone: 0049 (0)40 7410 26301

Dr Anton Davydok
Institut für Werkstoffphysik, Helmholtz-Zentrum Hereon
Outstation DESY Notkestraße 85,
22607 Hamburg
www.gems.hereon.de

Composition of the tumor microenvironment and immune repertoire in pediatric ALK-positive anaplastic large cell lymphoma

Partners: Prof. Dr. Wolfram Klapper and Prof. Dr. Monika Brüggemann, UKSH Campus Kiel

Anaplastic lymphoma kinase (ALK)-fusion protein-positive malignancies such as ALK-positive non-small cell lung cancer and ALK-positive anaplastic large cell lymphoma (ALCL) represent an ideal model forstudying tumor immunology: The ALK fusion protein is the driver of tumorigenesis, is almost exclusively expressed in the tumor cells and is recognized by the patient´s immune system. Pediatric ALK-positive ALCL patients mount an immune response against ALK characterized by the production of anti-ALK-antibodies and ALK-specific T-lymphocytes. The strength of this immune response is of high prognostic value. Little is known on the composition and competence of the immune cells in the microenvironment of pediatric ALCL. We propose a cooperative project together with the UKSH Kiel to (1) quantitatively describe the composition of the tumor microenvironment, (2) assess its functional capacity using gene expression analysis and (3) examine the T-cell receptor repertoire comparatively in blood and the microenvironment. Further elucidation of the host immune response against ALK-positive ALCL increases our understanding of the mechanisms behind the inter-individual strength of tumor-immunity. This forms the basis for the development of individualized therapeutic options like vaccination, check-pointinhibition or stem cell transplantation. Clinically, T cell receptor (TCR) repertoire analysis could represent a novel opportunity for long-term monitoring of patients.

Contact and affiliation:

Dr. med. Fabian Knörr
Research Institute Children’s Cancer Center Hamburg and Department of Pediatric Hematology and Oncology
f.knoerr@uke.de
+49 (0) 40 7410 51219

Prof. Dr. Monika Brüggemann
Department of Internal Medicine II, UKSH, Campus Kiel
Hämatologie Labor Kiel

Prof. Dr. Wolfram Klapper Institut for Pathology, Section Hematopathology, UKSH, Campus Kiel Institut für Pathologie Kiel

Immune determinants of melanoma metastasis to the brain

Despite recent advances in the treatment of advanced melanoma, MBMs remain a therapeutic challenge. Tothis day, the mechanisms of much worse prognosis for patients with MBMS than for patients with non-cranialmetastases remain unclear. We hypothesize that a less efficient anti-tumor immune response to MBMsaccounts for these different outcomes and propose to examine the systemic immunophenotype and localimmune reactions. Using existing blood samples from established repositories at the UKE, we plan on usingflow cytometry to describe the peripheral immune phenotypes of patients with MBM and compare these topatients with other distant melanoma metastases excluding the brain. Using existing tumor samples, we planon using regular immunohistochemistry, multiplex immunofluorescence and T-cell receptor sequencing toexamine the local anti-tumor immune response. This project proposal is a continuation of my previous researchand thesis work on primary brain tumors in the Department of Neurosurgery and establishes a newcollaboration between the Departments of Dermatology, Neurosurgery and Pathology.

Contact and affiliation:

Dr. med. Alessandra Rünger
Department of Dermatology and Venerology
In collaboration with “Labor für Hirntumorbiologie” – Department of Neurosurgery
a.ruenger@uke.de

The Role of innate (NK) and adaptive (T) effector lymphocytes in themicroenvironment of soft-tissue sarcomas: a comprehensive investigation usinghuman-derived sarcoma-organoids

Sarcomas are a group of heterogeneous malignancies rising from mesenchymal tissues. Despite theirrarity, a lack of sufficient systemic therapy strategies make sarcomas a challenging and clinically relevantsubject of basic scientific research6,7. While upcoming immunotherapies in different oncological fieldsshow promising results, up to this day, such approaches don’t play a relevant role in sarcoma-oncology8.With this project we would like to elucidate a new field of immunotherapy where it ironically got startedover a hundred years ago: in sarcomas9. To do so we aim to achieve a broader understanding of theimmunological microenvironment of sarcomas to investigate mechanisms which allow for immuneescape from both the innate and adaptive immune system. To study such mechanisms, we are currentlyanalyzing tumor infiltrating lymphocytes as well as cancer cells from patient-derived human sarcomasamples. We established patient-derived three-dimensional tumoroid models to evaluate humanlymphocyte-sarcoma interaction in vitro using the donors’ own tissue- and blood-derived lymphocytes.By expanding the understanding of lymphocyte-sarcoma interaction in those three-dimensional in vitromodels,we want to provide new approaches for immuno- and/or personalized sarcoma-therapy.

Contact and affiliation:

Dr. med. Adrian Sagebiel
Klinik und Poliklinik für Allgemein- Viszeral- und Thoraxchirurgie
a.sagebiel@uke.de

FrailPANC – Neoadjuvant treatment in frail patients with pancreatic adenocarcinoma – A prospective, randomized multi-centre Phase II AIO/ CHIR-Net Trial

The majority of patients with pancreatic adenocarcinoma (PDAC) presents itself around 70 years, often in a frail physical condition (ECOG ≥2). Still, most clinical studies neglect this important patient subgroup. The aim of the FrailPANC trial is to evaluate the role of neoadjuvant chemotherapy in frail patients with resectable or borderline resectable PDAC. The primary endpoint is the overall survival, followed by the overall treatment utility (OTU) as a co-primary endpoint. In order to enable the trial a short project proposal will be submitted to the Deutsche Forschungsgemeinschaft (DFG) by February 2020. If the proposal is positively reviewed, the full proposal will be prepared within the requested funding period of the MSNZ (estimated start 06/2020). During the funding period the recruitment of participating sites and the integration of translational projects will be addressed. One of the translational projects will be in cooperation with the Department of Tumour Biology at the University Medical Center Hamburg-Eppendorf (UKE) with regard to their expertise concerning circulating tumour cells, single cell analytics and establishment of tumour organoids in cooperation with the Dutch Pancreatic Cancer Study Group. A second translational project will be established with the Department of Anaesthesiology regarding their preliminary work within the perioperative care of the elderly patient. In case of rejection of the proposal by the DFG, we will submit a grand proposal to the Deutsche Krebshilfe (DKH) during the MSNZ funding period.

Contact and affiliation:

Dr. med. Mara Goetz
Department of General, Visceral and Thoracic Surgery
m.goetz@uke.de
+49 (0) 40 7410 54403

Studying the impact of tumor microenvironment and intestinal microbiota on CD4+ IL17+ heterogeneity in order to increase immunotherapy efficacy in colorectal cancer

Recently, immunotherapies targeting co-inhibitory receptors (CIRs) have revolutionized the treatment of many types of cancer. However, in colorectal cancer (CRC), the third most common cancer, immunotherapies did not meet the expectations. We hypothesize that two key factors contribute to the poor efficacy of the immunotherapy in CRC: (I) the functional heterogeneity of the targeted cells, (II) the local microenvironment. Our unpublished data show that non-conventional IL17A+ Foxp3+ CD4+ T cells promote tumor growth and express several CIRs. This has to be taken into consideration since this population of protumorigenic cells can be unleashed by immunotherapies obtaining the opposite effects: growth of tumors. By characterizing this pro-tumorigenic cell population, we seek to determine the best immunotherapy regimen which is able to selectively impair the activity of these cells in CRC patients. Furthermore, we seek to understand in what manner the composition of tumor invading intestinal microbiota shapes the expression of CIRs on these cells and thereby, directing the efficacy of immunotherapy.

Contact and affiliation:

Dr. med. Leonie Konczalla
Department of General, Visceral and Thoracic Surgery
l.konczalla@uke.de
+49 (0) 152 228 27 721

MATEO: Maintenance therapy vs. observation in FOLFIRINOX treated metastatic pancreatic ductal adenocarcinoma (mPDAC) patients - A prospective, randomized multi-center phase IIl AIO trial

For initiation of the project, a study synopsis will be finished with a description of objectives and endpoints, study design, methodology and statistical considerations. This synopsis will serve as a first proposal submission at DFG. Further preparations for the final grant applications include preparation of relevant study documents (protocol, patient information material, ethics approval), recruitment of study centers within the AIO (Arbeitsgemeinschaft Internistische Onkologie) and initiation of the organization and management of the trial. Additionally, the preparation of a trials in progress manuscript for a medical journal is planned. Complementing, the development of a concomitant translational project is planned with utilizing the ideal starting point of this study for translational research with a homogenous study cohort, prospective collected data and design with a treatment and non-treatment arm. We plan to monitor therapy responses with assessment of circulating tumor markers (CA 19-9) and circulating tumor DNA (ctDNA) as possible therapy predictors along with the routine clinical and radiological assessments to establish subgroups for future therapy choices. I personally will conduct the described preparation of the grant application and organization of the trial with its accompanying translational program with PD Dr. Marianne Sinn as a supervisor and leading principal investigator of the study.

Contact and affiliation:

Dr. med. Martin Schönlein
II. Medical Clinic and Polyclinic for Oncology and Hematology
m.schoenlein@uke.de
+49 (0) 152 228 72 093

Kinase activity profiles as prognostic marker for immunotherapy in head and neck squamous cell carcinoma

Immune checkpoint inhibitors (ICI) have already been approved as 1st and 2nd line therapy of relapsed or distant metastatic head and neck squamous cell carcinoma (HNSCC). However, only 13-18% respond to therapy with no definitive predictive markers available resulting in a great need to establish predictive tests identifying responding and non-responding patients. For ICI therapy of advanced melanoma it has been demonstrated, that specific kinase activity clusters of peripheral blood mononuclear cells (PBMC) can be predictive. These clusters result from functional kinome profiling and might be promising in HNSCC. To test this, liquid biopsies will be collected from HNSCC patients before and under ICI treatment. PBMC will be isolated and functional kinome profiling will be performed. After cluster analysis the kinase activity clusters of the PBMC will be compared with the survival data of the patients. This method may also allow to monitor the patients` individual response during ICI treatment enabling the early detection of a progresses. Such a predictive test would support to identify responding versus non-responding patients, guide non-responders to a more appropriate therapy. This enables a more efficient economical use of the expensive therapeutics. In the future, a predictive test may also be transferred to additional entities.

Contact and affiliation:

Dr. med. Lara Bußmann
Department of Otorhinolaryngology
l.bussmann@uke.de

Cholangiocarcinoma complicating PSC – which T cells are promoting cancer dissemination?

Cholangiocarcinoma (CCA) is a dreadful disease with a poor prognosis due to lacking therapeutic options, leading to death in 80 – 100 %. In Primary Sclerosing Cholangitis (PSC), a chronic and progressive inflammatory biliary disease, up to 40% of the patients develop CCA. The pathogenesis is poorly understood, but there is evidence that the immune system contributes towards promoting CCA. Histological studies have shown biliary intraepithelial metaplasia and dysplasia in PSC livers, suggesting an inflammation-driven metaplasia-dysplasia sequence CCA development Additionally, altered composition of the biliary microbiota from patients with PSC has been shown. The impact of the microbiota on T cell function - determined by the secretion of cytokines and expression of coinhibitory receptors - has been implicated in carcinogenesis. Thus, we hypothesize that protumorigenic T cells promote CCA development and dissemination. In order to define the functional phenotype, the co-inhibitory receptor profile, the origin of tumor infiltrating T cells and their interaction with tumor and biliary epithelial cells, we will perform single cell sequencing from blood, liver and tumor infiltrating T cells in combination with interactome analysis. Results will contribute to the understanding of the pathogenesis and reveal new immunotherapeutic options to directly serve patients suffering from this dreadful disease.

Contact and affiliation:

Dr. med. Jenny Krause
I. Medical Clinic and Polyclinic
je.krause@uke.de

Intratumoral heterogeneity of cancer stem cell features in hepatocellular carcinoma are decisive for indication of adjuvant treatment

The genomic and histopathological intratumoral heterogeneity of HCC (hepatocellular carcinoma) is of increasing translational interest. Dismal prognosis is frequently linked to high invasiveness and metastatic potential of HCC harboring CSC-features, indicated by aberrant EpCAM-expression. However, to what extent intratumoral distribution of CSC-features impacts on recurrence after curative treatment remains unknown. We hypothesize that only homogeneous positivity will lead to higher invasiveness, early dissemination and dismal outcome, identifying patients in urgent need for adjuvant treatment. Analog to colorectal cancer, high or low risk features for recurrence could be decisive for adjuvant treatment. This proposal aims to investigate the spatial heterogeneity of EpCAM-expression within HCC-nodules and its impact on a) local invasiveness, b) early dissemination, and c) clinical outcome measured by time-to-recurrence (TTR) and recurrence-free-survival (RFS). Complementary, we aim to decipher the mutational background of CSC-features, addressing the mutational profile of the most frequent oncogenic drivers1,2 and focusing on the non-canonical role of telomerase on Wnt/β-catenin signaling through activating TERT-promoter mutations. For our preliminary experiments we already designed a heterogeneity-TMA of 82 patients (341 spots). EpCAM-staining, data on CTC-detection and serum levels of AFP, AFP-L3, DCP as well as marker for Wnt/β-catenin activation have been completed. The mutational profile is expected for 01/2020.

Contact and affiliation:

Dr. med. Kornelius Schulze
I. Medical Clinic and Polyclinic
k.schulze@uke.de

Unraveling resistance mechanisms to immune-checkpoint inhibitors via molecular characterization of CTC in metastatic melanoma patients

Immune-checkpoint blockade therapy has revolutionized the treatment of patient with unresectable malignant melanoma. However, clinical outcome of the patients is obstructed by limited response rates and immune-related severe side effects. Therefore, there is an urgent but still unmet need for the identification of predictive biomarkers of clinical response. The aim of this project is to isolate circulating tumor cells from peripheral blood of metastatic melanoma patients treated by immune checkpoint inhibitors via a new enrichment method and to characterize them at the DNA and RNA level. Results from enumeration and molecular characterization will be correlated to clinical outcome and to other biological parameters in order to unravel diverse resistance mechanisms to improve the therapy regime. As melanoma is used as the prime model cancer entity for immunotherapy, this project will open new avenues for minimal invasive, blood-based, “liquid biopsy” biomarkers in other tumor entities (e.g., lung cancer) for which immunotherapy has also become a therapeutic option. This project is part of the LiquiMel Biobank which is organized by the applicant Julia Stadler and is a continuum of the UCCH fellowship of 2019. A close collaboration between the department of Dermatology and Tumorbiology supports the project.

Contact and affiliation:

Dr. med. Julia-Christina Stadler
aktuell: Klinik für Dermatologie
Medizinische Hochschule Hannover

stadler-j@outlook.de

Unraveling tumor dormancy in humans by autopsies and post mortem biopsies

In the cooperative research project between the Institute of Tumor Biology (ITB) and the Institute for Legal Medicine of the University Medical-Center Hamburg-Eppendorf (UKE), under the direction of Prof. Klaus Pantel and Prof. Klaus Püschel, I want to examine two scientific questions:

1. What is the percentage of the population carrying dormant tumor cells at the time of death without being diagnosed with cancer ? This would provide an estimate about the rate of the occult early dissemination of tumor cells before cancer diagnosis. Routinely obtained (anonymized) bone marrow as well as peripheral blood samples collected from deceased persons at the risk age of developing cancer (> 45years) will be analyzed for the presence of disseminated tumor cells (DTCs) and circulating tumor cells (CTCs).

2. Why are distant metastases in the skeletal muscles so rare even in patients with advanced cancer? Are CTCs not able to extravasate into muscle tissue or do DTCs exist in muscle tissue but remain in a dormant stage unable to grow out ? To investigate homing of CTCs to skeletal muscle (e.g. M. iliopsoas) originating from carcinoma patients, muscle samples of metastatic cancer patients (incl. solid tumors: gastro-intestinal and urogenital cancers, NSCLC and breast cancer, etc.) will be obtained during autopsies.

Contact and affiliation:

Dr. med. Antonia Fitzek
Institute for Legal Medicine
a.fitzek@uke.de
+49 (0) 40 7410 - 57840

Generation of an anti α and β chain chimeric antigen receptor (αβ CAR) to develop CAR-T cells and CAR-NK cells for therapy of T cell leukemia

Acute lymphoblastic leukemia (ALL) is the most common cancer in children. Even though ALL has a good overall prognosis, the more aggressive subgroup of relapsed T cell leukemia displays a poor outcome. Despite improvements in intensive chemotherapy and allogeneic hematopoietic stem cell transplantation (allo-HSCT), up to one third of pediatric T-ALL patients die of relapse or treatment failure. Therefore, new therapy options are urgently needed. In the last decades, immunotherapy became a turning point in cancer therapy. Antibodies targeting specific tumor antigens allow a highly specific and less toxic therapy. Moreover, cytotoxic immune cells modified with chimeric antigen receptors are able to kill tumor cells efficiently. Therefore, we aim to develop a chimeric antigen receptor targeting the alpha and beta chain of T cell receptors (αβTCR-CAR) to specifically destroy malignant T cells. Fratricide of CAR-T cells will be prevented by targeted disruption of the TCRβ locus.

Contact and affiliation:

Dr. med. Kerstin Schütze
Department of Pediatric Hematology and Oncology

The impact of hypo-fractionated locally ablative radiotherapy, surgery, chemotherapy, and radio-chemotherapy on the development of metastases in two prostate cancer xenograft tumor models

It is well known that radiotherapy can locally eliminate cancer cells. In addition to the local effects, radiotherapy also eliminates distant cancer cells, the so-called abscopal effect. However, its pathophysiological basis is not well understood and there is no information available on how to induce abscopal effects of radiotherapy that also influence the numbers of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs). In recent experiments with two small cell lung cancer xenograft tumor models we could demonstrate that hypo-fractionated high dose radiotherapy can induce such abscopal effects lowering the numbers of CTCs and DTCs. But the mechanism underlying these observations are very poorly understood. Hence, we want to expand our experimental database with different prostate cancer xenograft models to investigate whether abscopal effects can also be seen in this additional clinically important tumor entity and how cells of the innate immune system are influenced by radiation.

Contact and affiliation:

PD Dr. med. rer. nat. Thorsten Frenzel
Outpatient Center and Department for Radiotherapy and Radiation Oncology
+49 (0) 40 7410 54031
frenzel@uke.de

Relevance of SWI/SNF chromatin remodeling complex for gene expression and for regulation of transcription factors in human cancer - its role in the pathogenesis of acute myeloid leukemia as a model of malignant transformation in general -

The aim of this project is to characterize the relevance of the so called SWI/SNF complex for gene expression and for regulation of transcription factors in human cancer. This complex is a chromatin remodeler, consisting of about 20 different proteins, acting as a destabilizer of histone-DNA interactions and thereby changing transcription activity of several genes. SMARCA4 (SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4, synonym: BRG1) is the central ATPase containing enzyme and of upmost interest for this project because it is well known to mediate tumor suppressive effects in solid tumors. However, in the setting of acute myeloid leukemia (AML), the basic field of research of this project, SMARCA4 seems to play a different role, acting as a proliferation stimulus for myeloid blasts making it an interesting target for new therapeutic approaches. The examination of its function via in vitro and in vivo analysis in combination with clinical data assessment in hematopoiesis and pathogenesis of AML could give deeper insights into involved pathways and SWI/SNF mediated function in tumorigenesis, proliferation and dissemination. AML could serve as a model for malignant transformation in general, extending the gained data to several other tumor entities.

Contact and affiliation:

Dr. med. Franziska Modemann

II. Medical Clinic and Polyclinic for Oncology and Hematology
f.modemann@uke.de

Characterization of the liquorigenic seeding and identification of metastatic risk factors in medulloblastoma with morpho-molecular profiling and machine learning

Partner: Prof. Dr.- Ing. Andreas Schlaefer, TU Hamburg

Medulloblastoma is the most frequent malignant pediatric brain tumor. Metastatic dissemination occurs primarily via the cerebrospinal fluid (CSF). Accurate detection of metastases including microscopic disease in the CSF is key for an adequate therapy stratification. However, this is a difficult and time-consuming task and cytological examination is frequently unreliable. Here, we propose to establish a machine learning pipeline to automatize and improve diagnostic evaluation of CSF samples. Further, we will perform an integrated morpho-molecular analysis to study the relation between morphological features of tumor cells in the CSF and molecular as well as clinical properties of the primary tumor.

Contact and affiliation:

Dr. med. Michael Bockmayr,
Research Institute Children’s Cancer Center Hamburg and Department of Pediatric Hematology and Oncology
+49 (0) 40 7410 52725
m.bockmayr@uke.de

Prof. Dr.-Ing. Andreas Schlaefer
TU Hamburg, Medical Technology Science Systems

schlaefer@tuhh.de

Antiresorptive Drug-Related Osteonecrosis of the Jaw following systemic treatment for bone metastases – characterizationby Raman spectroscopy, histomorphometry and quantitative backscattered electron imaging

Antiresorptive agents such as bisphosphonates or the monoclonal RANKL-antibody denosumab are commonly administered in the treatment of bone metastases of solid tumors or multiple myeloma. They halt osteolysis by inhibiting osteoclast activity. A severe adverse effect is osteonecrosis of the jaw, frequently seen in oral and maxillofacial surgical patient care. After establishing a diagnosis, the treatment consists of surgical removal of necrotic bone while attempting to preserve viable bone. To date, suitable methods to accurately determine bone vitality remain deficient. Accordingly, patients are often confronted with recurrence of the disease, leading to subsequent further loss of jawbone with tremendous implications for ability of nutrition, speech, social interaction and overall quality of life. In this project, we introduce Raman spectroscopy to measure bone tissue viability, which is validated through high-resolution imaging methods. This merely optical, non-invasive technique is a promising candidate to ameliorate both diagnosis and treatment of patients affected by this debilitating disease.

Contact and affiliation:

Dr. med. Levi Matthies
Department of Oral and Maxillofacial Surgery
l.matthies@uke.de
+49 (0) 40 7410 - 53259

Understanding the local tissue quality changes within human spinal metastases of breast and prostate cancer

The underlying regulatory mechanisms of impaired bone matrix quality in metastatic spinal disease are poorly understood. Furthermore, its relation to metastatic growth, increasing the risk of paralysis due to spinal cord constriction (Figure 1) and fragility fractures, are not entirely elucidated yet. When cancer cells settle into bone, osteolytic or osteosclerotic lesions are caused by manipulating the activity of the bone-forming osteoblasts and bone-resorbing osteoclasts, at the same time benefiting from factors normally highly secreted by osteocytes i.e. sclerostin and RANKL via the metastatic cells. Bone stores growth factors and minerals that, when released, contribute to tumor progression. Bone matrix turnover is regulated by the mechanosensitive osteocyte network. This network of osteocytes might act as a central player, since I) osteocytes are regulators of bone remodeling, are II) actively participate in remodeling, and III) with their vastly active surface and interconnectivity could be the source of tumor cell-anabolic stimuli. We have designed a translational, interdisciplinary project utilizing a multitude of biological and material science techniques on precious human bone specimens of cancer patients. In this project we aim to assess the role of osteocytes in spinal bone metastases of breast and prostate cancer.

Contact and affiliation:

Dr. med. Annika Heuer
Department Trauma and Orthopedic Surgery
ann.heuer@uke.de

LiquidSWITCH – Early detection of resistance to BRAF inhibition and switch to immune checkpoint inhibition triggered by serial monitoring of cell-free DNA in blood plasma of melanoma patients.

Signal transduction inhibition as well as Immune-checkpoint inhibition (ICI) have revolutionized the treatment of patients with metastatic melanoma. However, patients with metastatic melanoma harboring BRAF mutations are still first treated with targeted therapy (TT) consisting of combined BRAF- and MEK-inhibitors until acquired resistance develops, and then switched to ICI. A radiological assessment following RECIST 1.1. criteria is the current standard definition of resistance. Nevertheless, the subsequent switch to ICI can be too late to improve patient overall survival. Therefore, there is an unmet need to identify predictive biomarkers of early resistance to TT. The aim of this project is to establish a biobank und analyze ctDNA and circulating tumor cells (CTC) from patients with non-resectable metastatic melanoma via digital droplet PCR (ddPCR) and next generation sequencing (NGS) and to characterize them at the DNA and RNA level. Within the framework of a multicenter, observational study four collaborating skin cancer centers in Northern Germany (UKE Hamburg, UKSH Lübeck, UKSH Kiel, Elbekliniken Buxtehude) will participate in patient recruitment. The analyses will be performed at the ITB (UKE) in collaboration with the UKSH Lübeck. Proof-of-principle data from this study will provide the basis for follow on studies with sufficient power to assess whether tumors responding to BRAF inhibition as defined by response in ctDNA can improve efficacy of immune therapy and vice versa.

Contact and affiliation:

Dr. med. Isabel Heidrich
Department of Dermatology/ Institute for Tumor Biology
i.heidrich@uke.de
+49 (0) 1522 - 2831042

Radiosensitization through molecular targeting in HNSCC tissue slice cultures of primary tumors and lymph node metastases

Curative intended radiochemotherapy in human papillomavirus (HPV)-negative head and neck cancer, especially when presenting with lymph node metastasis, is associated with high rates of locoregional failure and tumor relapse. This demonstrates the necessity for more efficacious concepts. Benefits made in precision cancer medicine could complement radiotherapy but the interplay of targeted agents with radiation is mostly unknown. Considering HNSCC heterogeneity and the lack of predictive biomarkers, patient-derived short-term assay systems may close this gap. In this project, we assess the potential of HNSCC tissue slice cultures of primary and lymph node metastasis as patient-specific tumor models for pretherapeutic testing of potentially radiosensitizing agents. Due to previous work we focus on two promising strategies: 1. Inhibition of hyperactivated src family kinases (SFK) and 2. Targeting of the cellular DNA damage response (DDR) and DNA repair machinery through dual inhibition of PARP and Wee1.

Contact and affiliation:

Dr. med. Henrike Zech
Department of Otorhinolaryngology
+49 (0) 152 22824064
h.zech@uke.de

Deciphering the Role of the Lymphotoxin – IL-22BP Axis in CRC metastasis

We have recently described how IL-22BP (interleukin-22 binding protein), a natural antagonist of interleukin-22, bridges the antitumorigenic effects of lymphotoxin in colorectal cancer (CRC) in mice and humans (Kempski*, Giannou* et al., Gastroenterology, 2020). Furthermore, we have shown that IL-22 promotes the first steps of metastasis formation. Our hypothesis is that IL-22BP is upregulated at later stages and controls the progression of already established metastasis. Preliminary data indeed show a dysregulated expression of IL-22BP in both the primary tumor of patients with metastasized CRC and in the liver metastasis themselves. However, the cellular source and regulation of IL-22BP in liver metastasis remain unknown. Our preliminary data indicate that the major producer of IL-22BP in metastasis is a different DC subset compared to the primary tumor in the intestine. Our next aim is to examine whether lymphotoxin regulates IL-22BP expression in this DC subset. Finally, we want to study whether the pharmacological inhibition of lymphotoxin signaling or the dendritic cell-specific knock-out of RelB (part of the non-canonical NFkB signaling pathway activated by lymphotoxin) can influence liver metastasis progression and might thus represent a novel therapeutic target for the treatment of metastasized CRC.

Contact and affiliation:

Dr. med. Jan Kemspki
Department of Internal Medicine I
+49 152 22829943
j.kempski@uke.de

Project 1: Intraventricular application of sonic-hedgehog pathway inhibition as a therapeutic option in young patients with medulloblastoma

Medulloblastoma is the most common malignant brain tumor in children, arises in the posterior fossa, and disseminates via the cerebrospinal fluid. One of 4 molecular medulloblastoma subgroups is caused by Sonic Hedgehog (SHH) overactivation. The small molecule Vismodegib allosterically inhibits Smoothened (SMO), the main upstream activator of SHH. Unfortunately, Vismodegib has shown to cause severe bone growth defects in preclinical studies and a phase II clinical trial, preventing its systemic application in children. Therefore, we aimed to establish an intraventricular therapy with Vismodegib, combining the benefits of targeted drug delivery to the location of the tumor and metastases and minimizing systemic side effects. In a mouse model for SHH medulloblastoma, we compared intraventricular, oral, and placebo treatment regarding effects on survival, tumor biology and bone morphology. Our results show a significant survival benefit of intraventricularly treated mice compared to placebo and no signs of systemic side effects on bone growth. Our experimental work is almost concluded, and we will now prepare a manuscript for publication.

Project 2: Molecular and clinical characterization of atypical peripheral nerve sheath tumors in NF1patients and prediction of risk for malignant transformation and dissemination

Neurofibromatosis Type 1 (NF1) is a tumor predisposition syndrome that causes a broad spectrum of benign and malignant tumors. Most frequent are neurofibromas (NF)– benign tumors that are divided into several different subtypes and that are associated with a variety of clinical complications (Philpott et al. 2017). Atypical neurofibroma (ANF) are believed to be at risk for progression to highly aggressive malignant peripheral nerve sheath tumors (MPNST) that have a dismal prognosis and are considered incurable upon occurrence of metastasis (Korfhage und Lombard 2019). Therefore, the identification of such lesions is of high importance for risk adapted patient care and could help to reduce the mortality of NF1 patients. Based on small series, different histological and imaging criteria have been proposed to identify ANF or “atypical neurofibromatous neoplasms of uncertain biological potential” (ANNUBPs), but a consensus pathological definition has not yet been reached (Miettinen et al. 2017). Most importantly, a thorough molecular and clinical characterization is missing completely to date. In this project, we aim to characterize the molecular, histological and clinical attributes that distinguish ANF with high potential of malignant transformation and metastasis from benign NF and malignant MPNSTs. To this end, we will perform in-depth histological analysis as well as global DNA methylation, copy number profiling, and sequencing of 30 ANF and compare these data with existing reference cohorts of NF and MPNST. Finally, we will investigate clinical parameters and outcome of ANF patients, including the type of NF1 germline mutation, patients’ age, tumor localization, malignant tumor transformation, potential metastatic spread and long-term follow-up. Respective results shall serve as a solid base for a tailored surveillance and future clinical trials for patients with ANF.

Contact and affiliation:

Dr. med. Catena Kresbach
Department of Neuropathology
Research Institute Children's Cancer Center Hamburg
+49 (40) 426051220
c.kresbach@uke.de