Full Proposal for the German Cancer Aid Priority Program 'Translational Oncology' (2st call) 2015 Lead Applicants: Prof. Dr. med. Magnus von Knebel Doeberitz Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg Prof. Dr. med. Jens Peter Klussmann Department of Otorhinolaryngology, Head and Neck Surgery University of Giessen Prof. Dr. med. Markus Löffler Institute for Medical Informatics, Statistics and Epidemiology (IMISE) University of Leipzig Coordinator: Dr. med. Miriam Reuschenbach Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg The Human Papillomavirus Infection and Oropharyngeal Cancer Consortium - HOCC

HPV-associated oropharyngeal cancer (OPSCC) HPV-associated malignant diseases cervix vulva vagina penis anus oropharynx skin o HPV in ~50% with increasing incidence o overall good outcome, but still cancer related death in 20% o substantial treatment-related morbidity → urgent need for therapeutics targeting the molecular characteristics of HPV-associated OSCC

HPV-associated oropharyngeal cancer (OPSCC) secondary alterations HPV E6/E7 overexpression Immunologic alterations Immune evasion Immunologic alterations Immune evasion Genomic alterations Activating mutations (oncogenes) HPV integration Genomic alterations Activating mutations (oncogenes) HPV integration Epigenetic alterations Methylation of viral regulatory regions Methylation of tumor suppressors Epigenetic alterations Methylation of viral regulatory regions Methylation of tumor suppressors

The Mission of HOCCTranslational relevance of the proposed research program HOCC Immune system (SP3) HOCC Genome (SP2) HOCC Epigenome (SP1) HOCC- Core (SP4) HOCC- Core (SP4) Immunologic alterations Activation of antigen-specific immunity, inhibition of immune evasion Trials with E6/E7, p16 INK4a vaccination; anti-PD-1 Immunologic alterations Activation of antigen-specific immunity, inhibition of immune evasion Trials with E6/E7, p16 INK4a vaccination; anti-PD-1 Genomic alterations Inhibition of activating pathways (e.g. PIK3CA) Trials on genomics-driven targeted therapy for HNSCC in general Genomic alterations Inhibition of activating pathways (e.g. PIK3CA) Trials on genomics-driven targeted therapy for HNSCC in general Epigenetic alterations Inhibition of DNA methyl transferases (e.g. decitabine) Phase I study initiated by applicants in Heidelberg NCT 3.0 program Epigenetic alterations Inhibition of DNA methyl transferases (e.g. decitabine) Phase I study initiated by applicants in Heidelberg NCT 3.0 program Validation of identified biomarkers and compounds in clinical trials. Evaluation of antigen- specific vaccination and immune checkpoint blockade in clinical trials. Concepts for combination therapies. Validation of prognostic and predictive markers. Identification of therapeutic biomarkers for DNA methyltransferase inhibitor treatment Identification of tumor subgroups responding to candidate compounds Identification of promising combinations of antigen-specific vaccination and immune checkpoint inhibitors Targets, interventions, status Deliverables of HOCC Subsequent steps

The Mission of HOCCThe structure of the proposed research program Whole genome methylation HPVE2BS methylation # Gene expression* # Whole genome methylation HPVE2BS methylation # Gene expression* # Test Cohort Patients with HPV+ OPSCC from existing cohorts (LIFE n=28 and HIPO-POP n=15), Test Cohort Patients with HPV+ OPSCC from existing cohorts (LIFE n=28 and HIPO-POP n=15), Validation Cohort Patients with newly diagnosed OPSCC, prospectively recruited, HPV+ n=150 Validation Cohort Patients with newly diagnosed OPSCC, prospectively recruited, HPV+ n=150 Methylation panel Therapy model Methylation panel Therapy model *data exist/generated alread from LIFE # data exist/generated already from HIPO Epigenome (SP1) Whole exome sequencing* # Copy number alterations # HPV integration Whole exome sequencing* # Copy number alterations # HPV integration Mutation panel Copy number alterations Therapy model Mutation panel Copy number alterations Therapy model *data exist/generated alread from LIFE # data exist/generated already from HIPO Genome (SP2) RNA sequencing Identification of antigens Antigen-spec. immunity Systemic and local immune profile RNA sequencing Identification of antigens Antigen-spec. immunity Systemic and local immune profile Antigen-specific immunity panel Systemic and local immune profile Antigen-specific immunity panel Systemic and local immune profile Immune system (SP3) integrative analysis HPV analysis data documentation material tracking integrative analysis HPV analysis data documentation material tracking HOCC-Core (SP4)

The Mission of HOCCSubproject Epigenome HOCC Epigenome (SP1) Objective 1: Establishment of methylation panels to enable biomarker-driven selection of patient subgroups for decitabine therapy Objective 2: Identification of therapeutic biomarkers to monitor efficacy of decitabine treatment Objective 3: Reciprocal effect between methylation patterns, the mutational landscape and the host immune system Identification of therapeutic biomarkers for DNA methyltransferase inhibitor treatment

The Mission of HOCCSubproject Genome HOCC Genome (SP2) Identification of tumor subgroups responding to candidate compounds Objective 1: Definition of genetic subgroups in HPV-positive OPSCC Objective 2: Development of a diagnostic NGS gene panel for patient risk stratification and identification of patient subgroups which benefit from PI3K/mTOR signaling pathway inhibition Objective 3: Validation of the NGS panel in preclinical model systems for drug response

The Mission of HOCCSubproject Immune System HOCC Immune system (SP3) Identification of promising combinations of antigen-specific vaccination and immune checkpoint inhibitors Objective 1: Identification of novel target antigens Objective 2: Analysis of humoral and cellular immunity against novel and known target antigens. Objective 3: Profiling of the functional activation state of circulatory and tissue-infiltrating immune cells

The Mission of HOCCSubproject HOCC-Core HOCC- Core (SP4) HOCC- Core (SP4) Objective 1: Establishment of a common consortium data and biomaterial base (cohorting) Objective 2: Coordinating the diagnostic and experimental work up (diagnostics and sample tracking) Objective 3: Conduction of integrative bioinformatic analyses (bioinformatics)

To directly bring the findings on HPV-associated OSCC into further clinical trials within the HOCC network Thereby complementing de-escalation trials in patients with HPV- associated OSCC And finally contribute to adequate treatment protocols for HPV- associated OSCC, which will become the dominant group of OSCC in the near future. The Mission of HOCCLong-term opportunities of the proposed program