Objectives of the proposal: Heart Failure (HF) is estimated to cost the EU economy €196 billion a year. Currently, patients are treated with the highest pharmaceutical doses of antiRAS agents, which cause severe side effects and reduce the patients’ quality of life. Moreover, treatment responses are highly variable. This study aims to enable imaging-guided treatment optimization by using a radiolabelled novel angiotensin 1 receptor (AT1) blocker analogue in a small animal model of myocardial infarction (MI). Positron emission tomography (PET) as a cutting-edge method of evaluating HF pathophysiology at a molecular level will be employed to optimize treatment. How the objectives will be achieved: MI will be obtained by ligation of the coronary artery in Wistar rats. Subsequently, PET imaging with a dedicated small animal system (mciroPET) will be performed. A multiple set of cardioneuronal PET tracers provided by the partner institution at Johns Hopkins University will be used to measure the extent of global and regional neurohumoral abnormalities in rat hearts following different degrees of ischemic insult. The relationship between neurohumoral system alterations and subsequent left ventricular remodelling will be determined by microPET and Magnetic Resonance Imaging to assess deterioration of ventricular geometric parameters. Using the novel AT1-ligand, imaging-guided dose-derived therapeutic concepts for Wistar rats will be established in vivo with the aim to transfer this technique to the clinic in a long term perspective. Relevance to the Work Programme: I envisage that in the future a single cardiac PET scan prior to treatment initiation will be sufficient to estimate the appropriate drug dose in any given HF patient. This new concept of individualized drug dose determination has the potential to lead to personalized treatment for one of the most frequent fatal and expensive diseases and thereby contributes to the reduction of the economic burden of the EU.

In 2020, the cell culture market was valued at $19 billion USD, with consumables comprising more than 60% of the market. The emerging 3D cell culture provides lucrative business opportunities for consumables and tissue test models, but current technology and market supply lack an easy and reliable tool for the uncomplicated creation and cultivation of vascularized and perfusable artificial 3D tissue. The current 3D cell culture consumable market offers only well plate inserts and organ-on-a-chip systems for generating 3D tissues, which are limited regarding tissue mimicry and complexity. Bioprinting, while promising, is still unreliable for reproducible vascularized tissue engineering. The EIC Transition Proposal Vasc-on-Demand aims to fill this gap by providing three easy-to-use laboratory products for the generation and cultivation of perfusable vascularized 3D tissue: 1. BasicVasc: An all-in-one bioreactor consumable for generation and cultivation of perfusable tissue culture 2. EasyVasc: Prefabricated ready-to-use vessel channel networks without cells 3. CompleteVasc: Prefabricated matured cell-containing vascularization EasyVasc and CompleteVasc both build on the BasicVasc technology. Therefore, BasicVasc will be advanced for high-throughput production of precisely manufactured structures to be ready for the market. The other products will be fully developed and characterized to allow improved vascularized tissue production with even more advanced and simpler ready-to-use systems. This grant will be carried out by a highly motivated team with synergistic expertise in technology, product engineering, and business development, working towards the foundation of a start-up for the commercialization of the technology. The proposed Vasc-on-Demand has the potential to simplify the development of sophisticated test models for research and pharmaceutical approaches like drug testing, leading to reduced animal trials and development costs.

Platelets are mediators of haemostasis and thrombosis, but also increasingly recognized as versatile effector cells of innate immunity. The concept of thrombo-inflammation recognizes that thrombotic and inflammatory pathomechanisms are closely intertwined. Platelets thereby act as central orchestrators of immune cell trafficking, vascular barrier function and organ integrity and contribute to organ injury in severe disorders with limited treatment options (e.g. stroke, sepsis, ARDS). The underlying molecular mechanisms are not known. I recently discovered an unexpected novel effector mechanism in platelets with potentially huge impact for understanding these cells in health and disease: Resting platelets can rapidly reorganize the entire pool of their principal adhesion receptor, integrin αIIbβ3, along with its associated tetraspanins and signalling machinery into ´disintegration´ complexes (DISCs) in distinct membrane microdomains. These DISCS serve as building blocks for a novel organelle, the ´Platelet-derived Integrin and Tetraspanin-enriched Tether´ (PITT). PITTs are loaded with signalling molecules, ribosomes and RNA and can segregate from the platelet to promote thrombo-inflammation. Also β1-integrins as well as glycoprotein (GP)VI can be integrated into DISCs and deposited at discrete adhesion points. I therefore postulate that circulating platelets have the capacity to use their principal adhesion/signalling machineries in two fundamentally different ways and thereby switch between the haemostatic and a thrombo-inflammatory effector programme. If proven correct, this would implicate a radically new paradigm in platelet biology, and open new avenues for the treatment of a wide range of diseases with major societal impact. PITT-Inflame will (i) provide a detailed molecular composition and architecture of DISCs and PITTs; (ii) decipher underlying signalling networks; (iii) identify PITT-induced effects on target cells and (iv) deduce therapeutic strategies.

Functional 3D tissue models could replace animal experiments and significantly reduce the cost of over 100 million € that pharmaceutical companies spend on failed drug development every year. The main bottleneck in the in vitro creation of 3D tissues is the need for perfusable vascularization in tissues bigger than 1 mm. So far, no generally applicable product exists for that purpose. This hampers advances in drug development as well as in research. Design2Flow will develop products, which overcome this limitation and enable the individualized on-demand creation of perfusable and therefore larger and more complex tissues. This will fundamentally reform research in life sciences by offering easy-to-use, customizable, perfusable 3D cell culture products of interest to the pharmaceutical industry and research labs. To achieve this goal melt electrowriting will be utilized to fabricate sacrificial scaffolds, which resemble the native microvasculature. The fabricated scaffolds will be combined with insert clips as easy-to-use designs for multiwell plates, which will allow a passive perfusion of the construct to keep the tissue alive without the need for a special pump – a beginner friendly way to start advanced 3D cell culture with basic laboratory equipment, without the need for expensive devices or experienced personal. For customers working with perfusion pumps, the proposed solution will be adapted to perfusion chambers for an active and controllable perfusion of the tissue by flow reactors, mimicking the in vivo blood flow even more accurately. This will provide an advanced and customizable way to create tissues as accurate as possible for physiological cultivation in bioreactors.

The oral epithelium is a unique tissue with a high degree of structural heterogeneity and distinct microenvironments (niches). However, the molecular mechanisms underlying the site-specific proliferation and differentiation of oral epithelial stem cells (OESCs) remain poorly understood. Oral squamous cell carcinoma (OSCC), one of the most common oral cancers, is a heterogeneous cancer type. Occurrences of metastatic lesions and treatment response differ from oral site to site, indicating a causal link to the heterogeneous nature of distinct OESC pools. In the OralNiche project, we will for the first time systematically and comprehensively characterise the OESC pools, dissect key mechanisms underlying oral epithelial site-specificity and define their contribution to OSCC heterogeneity. To achieve this, we will combine novel mouse models and patient material with cutting-edge methodology, including whole mount imaging, single-cell sequencing and organoids. Initially, we will profile the proliferative activities of OESCs and explore how stemness is regulated within defined niches in homeostasis, OSCC and chemotherapy-induced mucositis. Subsequently, we will functionally assess cellular cues that modulate stemness in the oral epithelia and generate a comprehensive single-cell atlas of OESCs and their cellular niches. Lastly, using OSCC patient material, we will validate key observations to define potential new biomarkers and therapeutic targets. In summary, this multidisciplinary approach will reveal how the distinct OESC pools maintain homeostasis, and how they respond to challenges, such as mucositis and OSCC. OralNiche will deliver new knowledge on the impact of tissue site-specificity on tumour heterogeneity and therapy response, which will have significant implications for OSCC patients. Moreover, the knowledge gained and the technological advances proposed will be applicable to other tissues and tumour types and thus provide a model approach in cancer research.