Dissecting cellular and molecular routes to pancreatic cancer phenotypes through genome-wide genetic screening in mice
Principal Investigator: Prof. Roland Rad (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM)
We propose to use transposon-technologies developed by us to perform genome-wide in vivo screens in different compartments of the mouse pancreas: acinar, ductal, endocrine and progenitor cells. This will (i) uncover molecular networks required for neoplastic transformation of each cell type; and (ii) unravel if and how the cell of origin and its mutational landscape affect molecular, histopathologic and clinical cancer phenotypes. We will functionally characterize new candidates in vivo using our novel CIRSPR/Cas9-based methods for somatic genome engineering. These studies will give comprehen-sive novel insights into the molecular and phenotypic complexity and plasticity of pancreatic cancer.
A porcine model of pancreatic ductal adenocarcinoma
Principal Investigators: Dr. Tatiana Flisikowska, Prof. Angelika Schnieke (Lehrstuhl für Biotechnologie der Nutztiere, Wissenschaftszentrum Weihenstephan, TUM)We will generate a model of human PDAC in pigs to translate advances in basic research into benefits for patients. The model is based on expression of inducible mutant TP53R167H and KRASG12D in pancreas. Pig lines carrying these genes have already been generated. Various means of activating pancreas-specific expression will be explored. The model will first be validated by comparison to the human disease, then further refined by establishing a pancreas organoid culture system for in vitro analysis, and efficient methods of cell-specific mutagenesis of PDAC-related genes in vivo and in culture.
Molecular determinants of acinar-ductal metaplasia and PanIN lesions
Principal Investigator: Prof. Roland M. Schmid (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM)In response to oncogenic signalling acinar cell undergo transdifferentiation to duct-like progenitor cell (acinar-ductal metaplasia (ADM)), form pancreatic intraepithelial neoplasia (PanIN) and progress to PDCA. Mice with oncogenic gain of function mutations of Kras, Pik3ca, Map2k1, and Braf develop ADM and PanIN with similar morphology. We will perform holistic studies to determine genetic, epigenetic, molecular, metabolic and signaling requirements to induce this highly plastic cellular state of ADM. The overall aim is to decipher the roadmap to ADM and PanIN and the heterogeneity of early lesions, which will help to understand PDAC subtypes with the potential of therapeutic intervention.
Functional characterization of the atypical member of the IkappaB inhibitor Bcl-3 in pancreatic ductal adenocarcinoma
Principal Investigator: Prof. Hana Algül (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM)The atypical member of the Inhibitor of kappa-B (IkappaB) family of proteins B-cell CLL/lymphoma 3 (Bcl-3) is an established oncogene in hematologic malignancies. Here we demonstrate for the first-time hyperphosphorylation of Bcl-3 in human and murine PDAC. Using various genetic tools we report that hyperphosphorylated Bcl-3 negatively regulates pancreatic carcinogenesis and metastasis. We will elaborate the previously unrecognized role for Bcl-3 and its phosphorylated sites in PDAC and highlight new aspects of Bcl-3 in this disease.
Dissecting Toll-like receptor 3 function in pancreatic regeneration and neoplastic transformation
Principal Investigators: PD Dr. Guido von Figura (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM), Prof. Bernhard Holzmann (Klinik und Poliklinik für Chirurgie, Klinikum rechts der Isar, TUM)The Toll-like receptor (TLR) pathways are frequently altered at the genetic level in PDAC. Based on this and our own preliminary work, which shows a defective pancreatic regeneration and pronounced Kras-driven inflammation in the context of TLR3 loss, we hypothesize that the nucleic acid sensor TLR3 plays an essential and compartment-specific role during regeneration and carcinogenesis in the pancreas. In this research initiative, we aim to dissect the specific role of TLR3 in these processes. The overall aim is to identify new druggable pathways that critically influence pancreatic tumorigenesis.
Defining and targeting subtype specific inflammatory drivers in pancreatic cancer
Principal Investigators: Prof. Dieter Saur (Chair of Translational Cancer Research & Klinik und Poliklinik für Innere Medizin II, TUM), Prof. Marc Schmidt-Supprian (Klinik und Poliklinik für Innere Medizin III, Klinikum rechts der Isar, TUM)
The molecular and cellular mechanisms linking the immune system with pro-tumorigenic signaling pathways in PDAC cells and their microenvironment are poorly defined. We will systematically dissect inflammatory signals derived from defined tumor subtypes and their microenvironment and investigate how these signals influence PDAC progression, therapeutic efficacy and treatment resistance. With our experiments, we aim to uncover targetable inflammatory drivers and develop novel treatment strategies for translation into the clinic in the future.
Functional characterization of neuropathy and neural invasion (NI) in genetically engineered mouse models (GEMM) of pancreatic ductal adenocarcinoma (PDAC)
Principal Investigator: Prof. Güralp O. Ceyhan (Klinik und Poliklinik für Chirurgie, Klinikum rechts der Isar, TUM)
The aim of our project is a comprehensive mechanistic understanding of the nerve-cancer interactions in PDAC. Neuroinvasion will be characterized in a panel of GEMM in vivo and in vitro to define genetic and signalling events mediating NI. PiggyBac transposon mutagenesis will be used as an unbiased approach to discover novel genes driving NI. Targets will be validated on human tissue cohorts and functionally tested using organoid-neuron co-cultures. The long-term goal is the development of therapeutic concepts interfering with NI.
Effects of obesity and anti-obesity treatments on pancreatic cancer
Principal Investigators: Dr. Kerstin Stemmer, Prof. Matthias H. Tschöp (Institute for Diabetes and Obesity (IDO), Helmholtz Zentrum München)
P08 aims to understand the impact of obesity, diet, caloric restriction and gastric sleeve surgery on formation and progression of PDAC. Mechanistic studies include the detailed characterization of inter-organ communication between adipose tissue and the tumor mediated by inflammatory adipokines and extracellular vesicles. The long-term aim is to develop novel prevention and treatment strategies tailored towards obese patients.
Identification of tumor-borne mediators of pancreatic cancer-associated cachexia
Principal Investigators: Dr. Mauricio Berriel Diaz, Prof. Stephan Herzig (Institute for Diabetes and Cancer (IDC), Helmholtz Zentrum München)
Cancer cachexia is a catabolic syndrome characterized by dramatic weight loss and tissue wasting that affects up to 80% of patients suffering from pancreatic cancer. Cachexia substantially impairs therapy response and worsens prognosis of affected subjects. The underlying mechanisms are yet unresolved and adequate biomarkers have not been defined. Furthermore, no curative treatment options are available in the clinics. Project P09 aims at identifying novel tumor-borne factors during cachexia, in order to define prognostic biomarkers as well as potential therapeutic targets for innovative treatment approaches.
Mechanisms promoting coagulation responses and thrombosis in pancreatic cancer metastasis
Principal Investigators: Prof. Bernd Engelmann (Institut für Laboratoriumsmedizin, Klinikum der Universität München, LMU), Prof. Steffen Massberg (Medizinische Klinik und Poliklinik I, Klinikum der Universität München, LMU)
The mechanisms triggering intravascular coagulation at sites of pancreatic cancer metastasis and those inducing cancer-associated deep vein thrombosis (DVT) are largely unknown. We will clarify the molecular circuits underlying tumor-triggered microthrombosis and reveal how such thrombi and extracellular vesicles control metastasis using intravital 2-photon fluorescence microscopy. In parallel, procoagulant reactions of immune cells during metastasis will be uncovered. The new molecular targets identified will be examined in models of DVT to enable specific inhibition of cancer-associated thrombosis.
Proteomics subtyping and circadian dynamics of PDACs to develop novel therapeutic strategies
Principal Investigators: Prof. Maria S. Robles (Institute for Medical Psychology (IMP), LMU), Prof. Dieter Saur (Chair of Translational Cancer Research & Klinik und Poliklinik für Innere Medizin II, TUM)
We will comprehensively study the proteome and phosphoproteome of genetically defined PDAC subtypes over time to achieve mechanistic insights into the cellular metabolism and signaling pathway networks driven by the circadian clock and different oncogenic mutations such as KrasG12D, KrasG12D/Trp53R172H, BrafV637E and Pik3caH1047R. The functional characterization of PDAC subtype specific candidate targets obtained from our proteomics/phosphoproteomics and circadian studies will allow us to pinpoint cancer markers and novel drug targets with the goal to develop subtype-specific chronotherapeutic treatment strategies for this devastating disease.
Targeting epithelial plasticity in pancreatic cancer
Principal Investigators: Dr. Maximilian Reichert (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM), Dr. Christina Scheel (Institute of Stem Cell Research (ISF), Helmholtz Zentrum München)
Project 12 presents a multi-pronged experimental strategy to target master regulators of epithelial plasticity in PDAC in vitro and in vivo using pancreatic cancer patient-derived organoids (PDOs). In work package 1 (WP1), the investigators will define a plasticity score. In WP2 and WP3, selected plasticity genes will be genetically or pharmacologically targeted followed by functional analysis. Using PDO-xenografts (PDOXs), promising candidates will be tested in vivo (WP4). The goal is to gain powerful mechanistic insights that enable the direct targeting of epithelial plasticity in pancreatic cancer patients.
Development of epigenetic-based therapies for PDAC
Principal Investigators: PD Dr. Günter Schneider (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM), Prof. Gunnar Schotta (Adolf-Butenandt-Institut, LMU)
Targeting epigenetics is a promising therapeutic concept. We want to follow two leads to develop new epigenetic therapies. Firstly, we will define Histone deacetylase (HDAC) 1, 2, 3 isoenzyme specific functions using novel genetic models. In combination with pharmacological screens we aim of developing novel HDAC inhibitor-based therapies. Secondly, we want to identify essential PDAC driver genes based on a specific epigenetic signature (super-enhancers). Mechanisms leading to forced expression of these driver genes will be identified through mapping of open chromatin regions within the super enhancers. Goal is to translate key findings into therapeutic concepts.
The biological role of Sphingolipids in pancreatic cancer
Principal Investigator: Prof. Julia Mayerle (Medizinische Klinik und Poliklinik II, Klinikum der Universität München, LMU)
Sphingolipid metabolism regulates cell death, differentiation, senescence, autophagy and migration. Employing metabolomics we delineated a metabolic signature discriminating between chronic pancreatitis and pancreatic ductal adenocarinoma (PDAC) suggesting a role of sphingolipid metabolism in PDAC development. Within this project, we will characterize the role of a key enzyme of sphingolipid metabolism for PDAC progression, metastasis formation and therapy resistance using genetically engineered mouse models. The long-term aim is the mechanistic understanding and therapeutic targeting of sphingolipid metabolism in PDAC.
Identification of biomarkers to predict individual radiosensitivity using pre-clinical pancreatic cancer models and development of innovative chemo-radiation therapy concepts
Principal Investigator: Prof. Stephanie Combs (Klinik und Poliklinik für RadioOnkologie und Strahlentherapie, Klinikum rechts der Isar, TUM)
Standard radiotherapy treatment of pancreatic cancer patients is performed without considering the individual tumor radiosensitivity, although differences in intrinsic radiosensitivity exist. The screening of radioresistance of murine PDAC cell lines and human organoids and the detailed information on the genome, transcriptome, proteome and phosphoproteome as well as the drug response will contribute to the identification of predictive biomarkers for radioresistance and the development of innovative treatment concepts.
Radioimmunotherapy of pancreatic cancer: From target validation to treatment response
Principal Investigators: Prof. Kirsten Lauber (Klinik und Poliklinik für Strahlentherapie und Radioonkologie, Klinikum der Universität München, LMU), Prof. Max Schnurr (Abt. für Klinische Pharmakologie, Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, LMU)
Pancreatic ductal adenocarcinoma (PDAC) is characterized by strong resistance against radio/chemo-therapy and potent immunosuppression. We propose to combine radiotherapy with the administration of bi-functional 5’ triphosphate siRNAs (ppp-siRNAs), which allow silencing of crucial DNA damage response (DDR) regulators and activation of retinoic acid inducible gene I (RIG-I), as a strategy to (i) sensitize PDAC to irradiation-induced cell death, (ii) activate intra-tumoral type I interferon (IFN) production, and (iii) stimulate adaptive anti-tumor immune responses.
Neo-epitopes derived from the mutated tumor suppressor gene RNF43 as targets for T cell therapy in pancreatic cancer
Principal Investigators: Prof. Markus Gerhard, Prof. Dirk H. Busch (Institut für Medizinische Mikrobiologie, Immunologie und Hygiene, TUM)
Adoptive transfer of autologous T cells genetically modified to express tumor-specific T cell receptors (TCRs) can mediate potent anti-cancer effects, particularly when neo-antigens are targeted, which also enhances the specificity of the therapy. Within this project, we aim at employing a new class of neo-epitopes derived from frameshift mutations in the tumor suppressor gene RNF43 for the treatment of pancreatic cancer. For this, we will predict and validate RNF43 neo-epitope targets, isolate TCRs specific for those targets from healthy donor repertoires, screen their in vitro functionality and finally investigate their therapeutic capacity in murine models of pancreatic cancer.
Prediction of drug sensitivity and resistance of pancreatic cancer cell lines
Principal Investigator: Prof. Bernhard Küster (Lehrstuhl für Proteomik und Bioanalytik, Wissenschaftszentrum Weihenstephan, TUM)
The central hypothesis of the project is that the overall signaling state of a pancreatic cancer cell is an important determinant for drug sensitivity or resistance and that the integration of proteomic and phosphoproteomic data with genetic information on pancreatic tumors and cell lines is a more powerful predictor of phenotypic drug sensitivity than any one approach alone. The 12 year vision is to use molecular profiling data of pancreatic cancer cell lines, organoids, spheroids and tumors to characterize their respective signaling pathways enabling the definition of subgroups and to develop subgroup-specific therapies.
Experimental resource and service platform
Principal Investigators: Prof. Roland Rad, Dr. Maximilian Reichert, PD Dr. Günter Schneider (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM), Prof. Dieter Saur (Chair of Translational Cancer Research & Klinik und Poliklinik für Innere Medizin II, TUM), Dr. Katja Steiger, Prof. Wilko Weichert (Institut für Allgemeine Pathologie und Pathologische Anatomie, TUM)
Limited accessibility of cutting edge model systems, insufficient characterization of therapeutic vulnerabilities, and the long time frames needed for functional in vivo studies are major bottlenecks of translational PDAC research. Therefore, service project S01 will provide standardized and quality-controlled multi-omics characterized murine and human in vivo and ex vivo models, core histopathologic analyses and patient cohorts, a drug-screening platform to decipher pharmacogenomics vulnerabilities, and an in vivo CIRSPR/Cas9-based rapid somatic pancreatic genome-engineering platform to support and accelerate individual research projects.
Cancer genomics, sequencing, informatics and data integration platform
Principal Investigators: Prof. Klaus A. Kuhn (Institut für Medizinische Informatik, Statistik und Epidemiologie, TUM), Prof. Roland Rad (Klinik und Poliklinik für Innere Medizin II, Klinikum rechts der Isar, TUM)
S02 is a platform for cancer genomic analyses that will provide sequencing and bioinformatics support, covering a broad spectrum of applications needed by subprojects. A second aim of S02 is to characterize the genomic landscapes of mouse and human PDAC cell line and organoid resources; and to integrate these genomic analyses with functional and phenotypic data, such as drug screens, histopathology or clinical/prognostic information. These cellular and data resources will be a critical experimental platform for CRC groups to dissect the molecular basis of various phenotypes. All data will be curated and made accessible through an intuitive data analytics and visualization platform.