ECRT Projekte

Das ECRT will insbesondere den wissenschaftlichen Nachwuchs beim Aufbau eigener Forschungsvorhaben unterstützen. Dafür bietet das ECRT Einstein Kickboxen mit einer Startfinanzierung für erste Experimente zur Validierung der Forschungsideen sowie ECRT Research oder Consumable Grants zur Weiterentwicklung der Forschungsvorhaben.

Die Zusammenfassungen der Projekte stehen nur auf Englsich zur Verfügung.

Sie befinden sich hier:

Frühere Einstein Kickbox Projekte

Einstein Kickbox - Förderperiode 2020

Extracellular matrix formation in pro- versus anti- inflammatory milieus – a key modulator in bone regeneration?

Team: Sophie Görlitz, Lisa-Marie Burkhardt, Aaron Herrera, Julia Berkmann, Christian Bucher

Funding: Einstein Kickbox - Young Scientists

The outset of all regenerative processes is defined by an inflammatory reaction. Inflammatory cells remove necrotic tissue, release inflammatory and proangiogenic factors and exert chemotactic effects. A tight regulation of the immune reaction is a prerequisite for an undisturbed healing. A prolonged pro-inflammatory reaction and an imbalance towards pro-inflammatory T cells impaired bone healing, while the transfer of anti-inflammatory T cells supported healing. Consequently, a bivalent influence of pro-/ anti- inflammatory T cells on fracture healing is suggested. Besides the known chemotactic and angiogenic functions, recent work describes a novel role of T cells in regulating extracellular matrix formation during bone healing. It was suggested that T cells help to organize the collagenous matrix and slow down the process of cartilage mineralization. Here we hypothesize that negative or positive effects of pro- or anti- inflammatory T cells, respectively, are at least partially related to modulations of ECM deposition. The link between T cells and ECM formation is however still poorly understood.

Hence, in this study we are planning to further dissect the influence of pro-/anti-inflammatory cytokines on extracellular matrix formation in a well-characterized in vitro 3D tissue formation model.  We aim to identify target cytokines which hinder or promote ECM formation. The expected insights are of high relevance for a better understanding of the crosstalk between the immune system and early ECM formation processes in bone healing and might contribute to develop future therapeutic concepts.

Pharmacologic inhibition of CRLR as a novel approach to treat obesity

Team: Jessica Appelt, George Soultoukis, Denise Jahn, Paul Köhli

Funding: Einstein Kickbox - Young Scientists

Obesity is a worldwide epidemic that has developed into a major global health problem affecting all ages and socioeconomic groups. This condition is often associated with an increased risk of stroke, diabetes mellitus type 2 as well as a reduced healing capacity. Interestingly, a weight reduction of only 5-10% can circumvent severe health conditions such as diabetes. It can be achieved through changes in diet, exercise or bariatric surgeries. However, adjustments in lifestyle often fail and surgeries are associated with increased risks for obese patients. Additionally, to date only a small number of drugs have been approved for clinical use. Thus, there is a demand for alternative strategies such as the development of efficient and safe anti-obesity drugs.

In this regard, a group of small peptides derived from the Calca gene may be of interest as targets for anti-obesity therapies – Procalcitonin (PCT) and alpha Calcitonin gene related peptide (αCGRP). Recently, we showed that mice lacking PCT and αCGRP kept on a high fat diet have a significantly improved health status compared to wild-type controls. Furthermore, both peptides are suggested to bind to the calcitonin receptor like receptor (CRLR), a G-protein coupled receptor and therefore an excellent drug target.

Hence, in our study we are planning to analyse the eligibility of CRLR antagonists as possible strategy for the fight against obesity, associated comorbidities and the related drop in regeneration capacity.

Are they in contact? - Image evaluation of the interfaces between different materials

Team: Ana Prates Soares, Andreia Sousa da Silveira, Heilwig Hinzmann

Funding: Einstein Kickbox - Young Scientists

In biomedical sciences, the interaction between organism and biomaterials is defined at the interface. When body and material enter in contact, they react to each other. As a result, rejection, absorption, oxidation, bonding, integration, failure or success can occur. The amount of contact between materials can be a good parameter for evaluating how stable their attachment is. Our aim is to create an image-computational tool that can calculate and display the contact surfaces between materials and tissues. In the present project, non-destructive imaging data from classical X-rays, Cone Beam Computed Tomography (CBCT), Micro-CT and Phase contrast-enhanced micro-CT (PCE-CT) will be used. We will develop an image-computational tool to assess the contact at the interface between materials from grayscale images to obtain a statistical quantification of interfacial contact. The output of the proposed approach will reveal detailed information about the superficial area of contact between the biomaterial and hard tissue. In the future, the use of our tool may help getting better insights about the correlation between interfacial contact and biomechanical properties. Thus supporting the development of new materials and design optimization.

iPSC-Derived Cardiac Progenitors - Who Are You?

Team: Ana Garcia Duran, Timo Nazari-Shafti, Sebastian Neuber, Andranik Ivanov

Funding: Einstein Kickbox - Young Scientists

The regenerative potential of the heart is not sufficient to repair damage caused by myocardial infarction. Thus, there is an urgent need for approaches that reverse cardiac remodeling, e.g. by providing a pro-regenerative environment or generating de novo heart muscle cells. Research on multipotent cardiac progenitor cells (CPCs) has gained traction as they can differentiate into cardiomyocytes, endothelial cells, and smooth muscle cells in vivo, potentially overcoming the limitations of current cell therapy strategies. In humans, various populations of CPCs, including Sca-1+, have been shown to improve cardiac function in preclinical models of myocardial infarction, contributing to the regeneration of the infarcted area. However, limitations associated with these cells, particularly poor accessibility and ethical concerns, hamper clinical translatability. Human induced pluripotent stem cells (hiPSCs) represent a readily available source of autologous fetal-like CPCs. We found that Sca-1+ cells isolated from hiPSCs during cardiomyocyte differentiation exhibited a stable phenotype through in vitro cultivation and were similar to human fetal CPCs in terms of morphology and gene expression of cardiac markers such as GATA4 and Nkx2.5. Our aim is to further characterize Sca-1+ cells to gain a better understanding of different Sca-1+ subpopulations and to identify GMP-compliant isolation and expansion strategies using single-cell RNA sequencing and cell surface marker screening.

The influence of Short Chain Fatty acids and other metabolites produced by commensal bacteria on macrophage biology – a high throughput screening approach

Team: Elena von Coburg, Caitlin Jukes, Philipp Burt, Liviana Ricci

Funding: Einstein Kickbox - Young Scientists

Commensal bacteria play a hugely important role in maintaining gut health by producing essential vitamins and breaking down non-digestive carbohydrates. Previous studies have shown that the microbial communities and their metabolites provide crucial signals upon inflammatory insults that alter the immune response of the host. Of note, the microbiome itself and its metabolites can be altered in human diseases such as inflammatory bowel disease (IBD). It is thought that these metabolic changes can impact the phenotype of immune cells within the gut. Intestinal macrophages are highly sensitive immune cells reacting to environmental cues. They adjust their pro-inflammatory and anti-inflammatory properties to modulate immune responses and maintain mucosal homeostasis. Moreover, numerous studies have shown the anti-microbial impact of bacterial metabolites on macrophages and their pro-inflammatory properties in IBD patients. To date, it is unknown which bacterial species are able to drive this impact and whether this effect can be recapitulated in vivo. In our project, we aim to identify a common molecular signature that drives an anti-microbial phenotype in macrophages by combining functional bacteria killing assays with high throughput techniques for gene expression and cytokine secretion. This will allow us to rapidly screen bacteria, which have regenerative therapeutic potential based on their ability to ameliorate the pro-inflammatory macrophage phenotype found in inflammatory diseases.

Unravel fibronectin tensional state – how does 3D physiological microenvironment impact early ECM formation?

Team: Matthias Kollert, Georgios Kotsaris, Julia Mehl

Funding: Einstein Kickbox - Young Scientists

The interplay of cells with their surrounding matrix is essential in homeostasis, regeneration and diseases. Apart from cell morphology being affected by mechanical cues from their microenvironment, it was found that early native extracellular matrix (ECM) development affects cell function. The interactions of structural ECM components were shown to be mechanoregulated as cells mediate the remodeling of fibronectin (Fn) matrix into a predominantly collagen matrix.

Fn is one of the most abundant ECM proteins and exhibits different conformational states depending on local strains in early ECM development which directly affects ECM formation. For example, in vitro experiments in 2D systems revealed that the tensional state of Fn regulates the nucleation of other ECM proteins such as collagen I. Here, we want to use a physiological 3D microenvironment with viscoelastic material properties to investigate early ECM formation in vitro.

ECM is a dynamic network undergoing constant modifications during regeneration. Understanding the mechanoregulation of early ECM formation by elucidating the spatiotemporal tensional states of the Fn matrix could answer why Fn is crucial for the assembly of other ECM proteins and how it is affected by tissue mechanical properties. We propose a novel strategy combining 3D alginate hydrogels with Fn visualization techniques which will be beneficial to regenerative therapies and tissue engineering and could be extrapolated to model degenerative diseases.

Next generation CAR-T – A novel virus-free approach to generate safe and effective CAR-T cell products for third-party use

Team: Dimitrios L. Wagner, Jonas Kath und Leila Amini

Funding: Einstein Kickbox - Young Scientists

T cells are a major part of our adaptive immunity, being able to eliminate infected or transformed target cells and are increasingly exploited as powerful tools in oncology. They can be redirected to recognize and eliminate cancer cells using Chimeric Antigen Receptors (CARs), fusion proteins of antibodies for antigen recognition and T cell-specific signaling domains. While already effective in patients suffering from certain blood cancers, the generation of an individual autologous CAR-T-Cell product for a single patient is a difficult and costly endeavor. Treatment delay due to production processes and high costs hinder their broad application in the clinic. This kickbox application aims to test a novel approach to create CAR-T-Cell product from healthy donors that are unable to attack the patients cells via their endogenous T cell receptors in an allogenic treatment setting. Therefore, this "next generation" of CAR-T cells may enable the generation of potent but cost-effective treatments of cancers. 

Photo-crosslinkable cytokine-release system for enhanced bone regeneration

Team: Norman Drzeniek, Paul Köhli

Funding: Einstein Kickbox - Young Scientists

The only commercially available drug for the treatment of non-union of long bone fracture, local recombinant BMP-7, was taken off the market due to serious side effects, leaving orthopedic surgeons without any further pharmacological solutions for a challenging pathology.

Since then, IGF-1 has been investigated among other cytokines as a promising alternative for bone healing. However, the key limitation of cytokine therapy remains the short in vivo half life of just a few minutes and so far no alternative to BMP-7 made it to clinical application.

To address this issue we want to design a novel type of "in vivo drug factory" that would prolong the release of IGF-1 in vivo but could also be easily assembled during surgery and injected into the fracture gap. Our strategy involves the combination of mRNA-transfected hMSC as a transplantable source of IGF-1 with a biomaterial vehicle that would interact with cell-secreted cytokines and modulate their release kinetics.An additional engineering challenge lies in combining cell transfection with cell encapsulation and material injection into a simple procedure that respects the clinical setting of use.

The role of the circadian clock in bone healing

Team: Denise Jahn, Serafim Tsitsilonis, Achim Kramer, Johannes Keller

Funding: Einstein Kickbox - Advanced Scientists

Impaired bone healing occurs in up to 10 percent of fractures and leads to pain, a long-term reduction in the quality of life and high socio-economic costs. More than 50 years ago, clinicians observed the phenomenon of traumatic brain injury (TBI) accelerating fracture healing. Until now, the underlying pathophysiology remained unknown, but we previously found compelling evidence that the stimulatory effect of TBI on fracture healing is transmitted through an increased adrenergic signaling, caused by an interrupted circadian clock. Several studies have shown a circadian rhythm in bone metabolism, affecting osteoclast activity, osteocytes and osteoblast function. Furthermore, the circadian clock regulates the body physiology through the sympathetic nervous system, which has a strong impact on bone tissue. Although there are a number of studies investigating the influence of the circadian clock on bone metabolism, the impact on fracture healing is largely unknown. In our project, we would like to investigate the role of the circadian clock in bone healing. Therefore, we will use a mouse line with a disrupted circadian clock in our standard osteotomy model. In addition, we will monitor the circadian rhythm of these mice as well as the released catecholamine. The understanding of the role of the circadian clock in fracture healing holds great potential to aid in the development of new drugs to improve fracture healing.

Developing menstrual blood–derived mesenchymal stem cell transplantation as an anti-inflammatory and proregenerative therapy for osteoarthritis by silencing TLR2 expression

Team: Ping Shen, Max Löhning, Hans-Dieter Volk, Tobias Winkler, Tazio Maleitzke, Lisa Grunwald

Funding: Einstein Kickbox - Advanced Scientists

Osteoarthritis (OA) is a chronic joint disease featured by cartilage deterioration and chronic pain. Regardless of the complexity of the initial causes, chondrocytes residing in cartilage are exposed to endogenous TLR agonists that are generated during cartilage breakdown. For instance, the 32-mer and 29-kDa fibronectin fragment, enzymatic products of matrix proteins, have been detected in the synovial fluid of OA patients and revealed an anti-anabolic, pro-catabolic and pro-inflammatory function by activating TLR2-mediated signalling pathways. Mesenchymal stem cells (MSC) hold great promise as treatment for the inflammation-accelerated degenerative disease OA, as they simultaneously carry regeneration potency and immunomodulatory capacity. On the other hand, MSCs are sensitive and respond to the inflammatory and degenerative cues, such as TLR agonists in the diseased microenvironment, to secrete inflammatory cytokine IL-6, IL-8 and G-CSF. We plan to overcome this limitation by endowing MSCs with a resistance to the local TLR2 agonists by silencing TLR2 expression. We will use menstrual blood–derived mesenchymal stem cells (MenSC) which hold, among other benefits, the advantage of non-invasive and periodical acquisition. Thus, we will transplant TLR2ko MenSCs to the knee joints of the OA-diseased guinea pigs and evaluate whether the knockout of TLR2 will endow MenSCs with an advanced therapeutic effects comparing to wild type MenSCs.

Phenotype and transcriptional landscape of T cells in acute and chronic joint inflammation

Team: Caroline Peine, Nayar Durán, Tazio Maleitzke, Philipp Burt

Funding: Einstein Kickbox - Advanced Scientists

Osteoarthritis (OA) is a severe disease with a very high prevalence world-wide. Only recently, the involvement of an inflammatory component in OA was recognized. Inflammatory cell infiltrates are frequently found in the synovium and the adjacent infrapatellar fat pad of the affected joints. In addition, also joint traumata are associated with a phase of synovial inflammation and it is generally accepted that patients that have experienced such a trauma have an increased likelihood to develop OA in the respective joint later in their life. Intraarticular soft tissue inflammation can be detected very early in the course of OA and is correlated with more rapid cartilage destruction. Besides macrophages and neutrophils, infiltrates of T cells are found in OA-associated joint inflammation. Mouse models have shown a substantial contribution of CD4+ and CD8+ T cells to the development of posttraumatic OA. Hence, using flow cytometry, we will study which T cell subtypes are present in the inflamed intraarticular soft tissues early after traumatic events such as meniscal tears compared with those in late-stage OA patients undergoing knee-replacement surgery. Thus, we can potentially identify specific pathogenic T cell subpopulations that are present both in acute and chronic joint inflammation and that could present a potential therapeutic target for the early treatment of posttraumatic OA.

Models deciphering mechanical force-induced tenocyte-immune cell signaling

Team: Franka Klatte-Schulz, Birgit Sawitzki, Sara Checa Esteban, Georg Duda, Gäbel Christiane, Serafim Tsitsilonis

Funding: Einstein Kickbox - Advanced Scientists

Tendon pathologies are very common and represent a significant burden for the patient. The pathomechanisms of chronic tendon pathologies (Tendinopathy) as well as failed tendon regeneration after acute rupture are not fully understood. Therefore, to date there exist no treatment option to support tendon healing, or to prevent the development of tendinopathy.

Excessive mechanical forces can cause acute ruptures and recurrent mechanical overload induces micro ruptures that lead to chronic conditions. Furthermore, inflammation is crucial for tendon healing and the development of tendon pathologies. In autoimmune-driven Achilles tendon enthesitis, IL23 Receptor (IL23R) expressing γδ T-cells that secrete IL17 have a driving pathogenic role. Previously, we showed that IL17/IL23R signaling is also present in non-autoimmune tendon pathologies. We hypothesize that mechanical strain together with inflammatory signals can modify the tenocyte-immune cell communication.

We will use an ex-vivo model of mechanical force-induced tenocyte-immune cell communication and gather information on factors influencing the IL17/IL23R signaling. In addition to varying mechanical strain or time, we will test different inflammatory triggers and cellular compositions on the secretion of IL6/IL23/IL17 associated cytokines and MMP classes. The obtained results will be used to develop a mechano-biochemical mathematical model, which aims on investigating the spatio-temporal regulation of IL6/IL23/IL17 signaling under mechanical and inflammatory stimulation.

Clinically Usable Allograft Cell-based Assay for the Assessment of Alloreactive T cells in Kidney Transplant Patients

Team: Constantin Thieme, Nina Babel, Toralf Roch

Funding: Einstein Kickbox - Advanced Scientists

Suppressing unfavorable immunity towards the allograft and the complications that this immune suppression entails are main challenges in organ transplantation. Several immunosuppressive agents and protocols are currently in use and applied according to a rough risk assessment using the HLA-mismatch, the level of antibodies against alloantigens, and individual experience of the treating clinician. Thus, the initial treatment can be adjusted with the onset of complications due to too strong or too weak immunosuppression. However, complications harming the allograft are currently only detected when the graft function is already impaired, often leading to irreversible graft damage. This deteriorates the long-term graft survival. Therefore, there is an urgent clinical need for guidance of immunosuppressive therapy. The research project that we will execute with support of the Einstein Kickbox grand is based on a method established and patented in our group that uses kidney transplant cells collected and cultivated from urine of kidney transplant patients. We envision redefining our protocol in order to make it more simple and applicable in a clinical context. The Einstein Kickbox grant with the BioThinking support will enable us to generate ideas and hypothesis that facilitate the assay optimization under consideration of the clinical and diagnostic demands. It also will help to find support from funding institutions and potential collaboration partners in the industry.

Einstein Kickbox - Förderperiode 2019

Generating micro-hearts with disease-specific phenotypes as testing platform for cardiac regenerative therapies

Preclinical testing of cardiac regenerative therapies such as direct reprogramming of fibroblasts into cardiomyocytes (CM) usually starts in vitro, with factor screens in 2D-cultured healthy cells, and proceeds in vivo, with candidate evaluation in animal models of heart disease. 2D models insufficiently emulate the real situation, whereas 3D models more accurately mimic cardiac multicellularity and extracellular matrix (cECM). However, an in vitro 3D testing platform that bridges the gap between 2D models and the in vivo scenario by adequately modeling cardiac pathologies is not yet available.

Therefore, the main goal of the proposed project is to engineer for the first time μ-hearts that mimic acute myocardial infarction (AMI) and chronic heart failure (CHF) and to demonstrate their utility for therapy testing. To do so, uniformly-sized spheroid μ-hearts will be assembled from human iPSC-derived cardiac cells and cECM at disease-specific ratios and culture conditions. After viability confirmation and immunohistochemical characterization, in situ genetic CM reprogramming will be tested. Finally, contractile motion analysis and imaging mass cytometry (IMC) will be evaluated for outcome analysis.

In the future, using iPSCs from patients with genetic cardiac defects will render this innovative testing platform suitable for personalized medicine. Eventually this may help to accelerate therapy development and maximize its effect while reducing the number of animal experiments.

Einstein Kickbox Projects - Förderperiode 2018

A Quantitative Approach to ECM Function in Aged Muscle Regeneration – Towards Predictive Modeling of Cell-Matrix Interplay

Ageing affects the regenerative abilities of skeletal muscle resulting in compromised healing associated with fibrosis, impairing mobility and affecting quality of life. Current research is largely focused on intrinsic muscle stem cell function, but local extrinsic changes are mostly neglected. However, skeletal muscle repair relies on a dynamic interplay between muscle satellite cells (SCs) and the extracellular matrix (ECM) microenvironment. We have preliminary evidence for a transient developmental-like ECM during early phases of muscle regeneration. We believe that this creates a biomechanical three-dimensional micro-niche conducive to SC expansion, differentiation and self-renewal during regeneration, and that its derailment in aging or disease contributes to SC malfunction. However, the mechanical properties of the pro-regenerative ECM, especially during aging, has not been analyzed and the influence of the bio-mechanical properties of the aged ECM on stem cell behavior remains unclear. By combining experimental and mathematical analysis in an interdisciplinary approach, we aim to comparatively determine and quantify the dynamic spatio-temporal structure, composition and functionality of the transitory ECM in young and old mice. Iterative mathematical modeling via integration of biological data in a stepwise fashion will in the future develop a predictive landscape to achieve a new level of understanding of ageing processes prospectively transferable to muscle-degenerative disorders.

Team: Sigmar Stricker, Arunima Murgai, Georgios Kotsaris, Max von Kleist, Vikram Sunkara

Funding: Einstein Kickbox - Advanced Scientists

Alterations in endothelial cell mechanics affect nuclear morphology and chromatin conformation: the mechanogenomic code

In this project we aim to establish a new scientific research focus at the ECRT, mechanogenomics. Multiple redundant interactions connect the plasma membrane to the nucleus allowing for external mechanical forces to alter nuclear morphology, a process which has been shown to have influence on the positioning of chromatin within the nucleus. Chromatin that associates with the nuclear membrane, so called lamina-associated domains (LADs), contains mostly inactive genes. We therefore hypothesize that mechanical forces directly influence gene expression via re-positioning chromatin in 3D nuclear space. Alterations in the extracellular matrix (ECM) composition might therefore modulate gene transcription during disease and regeneration. Preliminary data with CRISPR/Cas9 edited BMP receptor mutant endothelial cells show striking changes in cellular mechanics, ECM and nuclear lamina composition, as well as increased F-actin bundling and tubulin acetylation, resulting in increased cellular tension and nuclear deformation. In this context, we suggest that altered BMP signaling accounts only for a subset of differential gene expression, while other genes are regulated as a consequence of nuclear adaptation to novel mechanical inputs. Here, we aim to characterize the nuclear modifications with classical biochemical techniques and combine this, together with genome-wide identification of LADs and the corresponding transcriptome. Understanding the changes in the underlying regulatory gene networks and the synergism between BMP and mechano-signaling would provide novel insights for basic science as well as opportunities to translate this knowledge towards future treatment options in the clinics.

Team: Petra Knaus, Stefan Mundlos, Michael Robson, Jerome Jatzlau

Funding: Einstein Kickbox - Advanced Scientists

Systems Biology pipeline for discerning the disease phenotype of chondrocytes in osteoarthritis of the knee

Osteoarthritis of the knee (OAK) is one of the top five most disabling conditions that affects more than one-­third of people over 65 years of age and over 100 million individuals globally. OAK is characterised by an erosion of the articular cartilage, which leads to inflammation and pain accompanied by joint stiffness and immobility. Despite its prevalence, the molecular mechanisms triggering OAK are poorly characterised and current therapies cannot halt-­ or revert OAK. Consequently, knee joint replacement is often the only rehabilitative intervention. While animal models of OAK exist, the causative mechanisms leading to cartilage erosion are poorly understood. It is often unclear which of the OA-­associated factors are causing the disease, and which are a consequence or a mere side-­effect of it. Backed by literature evidence and own preliminary results on chondrocyte dynamics, we hypothesise that a disease-­associated chondrocyte (DAC) phenotype exists in early OA, causing subsequent cartilage erosion. Thus, identifying and characterizing DACs would pave the way for personalized diagnostic and therapeutic strategies to tackle an unmet clinical need. Within this project, we will identify the transcriptome signature of DACs from a mixed sample (healthy + DAC + noise). To do this, sophisticated bioinformatics and mathematical tools will be developed and applied to new experimental data from early OA time points, ensuring that DACs are sampled. Candidate transcripts will be compared to previously reported early OA-­associated factors, which typically contain mixtures of causative and consequential factors.

Team: Vikram Sunkara, Annemarie Lang, Max von Kleist, Christoph Schütte, Birgit Sawitzki

Funding: Einstein Kickbox - Advanced Scientsits

Control of Human Tissue Homeostasis and Immunity by Helper T cells (TiSSueHeLP)

The human body is constantly challenged with manifold pathogenic, chemical and physical stimuli. One of the major tasks of the immune system is to distinguish between harmless and harmful challenges in particular at barrier sites such as the skin. How do local T cells in such situations sense “danger”, localize challenges at defined tissues or cells and finally dose and center a right suitable reaction? How are regenerative processes induced, and do they differ in the context of pathogen- versus non-pathogen-type challenges? It seems to be logical that while a virus-infected cell has to be eliminated by cytotoxic effector T cells, such a reaction would be unsuitable for cells that have been activated by harmless chemical or mechanical stress. The immune system has to tightly control any inexpedient cytotoxic T cell action in particular at barrier sites. However, it is so far not known, how the activation of non-cytotoxic T cells is regulated at barrier sites.

In this Einstein Kickbox application we aim to perform multidimensional cytometric and histological analyses on circulating and tissue resident T cells to decode and delineate the human universe of T cell subsets according to novel cell surface signatures we have identified in our prework. Based on the achieved results we aim in further future activities to characterize the function of non-cytotoxic helper T cells in human tissues in particular in response to non-pathogenic stressors.

Team: Andreas Thiel, Christos Nikolaou, Lucie Loyal, Kerstin Wolk, Demetrios Christou, Roland Lauster

Funding: Einstein Kickbox - Advanced Scientists

3D - ALL - The tumor microenvironment of ALL in a bone marrow model

The aim of the project is to set up an as possible true-to-life model to study the microenvironment of primary acute lymphoblastic leukemia (ALL) cells on cell survival and resistance development. It is planned to investigate the effect of different drugs especially on the microenvironment but also on the ALL cells. While most cancer research focuses mainly on the pharmaceutical impact on tumor cells, the approach of this project will concentrate on the bone marrow stroma. We plan to establish a co-culture of ALL cells with 3D cultured bone marrow Mesenchymal Stroma Cells (BM-MSCs), as published by Sieber et al. (2017). For that, a hydroxyapatit coated zirconium oxide ceramic is seeded with patient derived MSCs and ALL cells to mimic the in vivo conditions. The whole cultivation is performed dynamically in the "Multi-Organ-Chip" (MOC). The MOC-platform, which has been developed at the chair of Medical Biotechnology at Technical University, is a microfluidic device consisting of a circular channel system which connects wells for the culture of different small functional human organ units, called organoids. Currently used 2D stroma cultures fail in their ability to support primary ALL cell proliferation and are therefore limited to test regenerative potentials of MSC cultures. This minimized model of ALL could be used for drug testing and dosage estimation. The in vitro leukemia model would present a breakthrough for detailed studies of the human marrow and for personalized drug screenings.

Team: Kübrah Keskin, Tessa-Lara Skroblyn, Domenic Schlauch, Cornelia Eckert, Mark Rosowsky

Funding: Einstein Kickbox - Young Scientists

Linking functional outcomes with regenerative potential: Can ultrasonography identify biomarkers of early healing and long-term recovery in human Achilles tendon rupture?

The ability to walk independently and pain free directly affects mobility and quality of life1,2. An essential function involved in walking is foot plantarflexion, articulated by a muscle-tendon unit containing the Achilles tendon (AT). Acute injury, like Achilles tendon rupture (ATR), leads to lowered strength and long-term functional outcomes3–5 regardless of treatment6, and puts patients at a higher risk for further injury7,8. The incomplete regeneration of the AT is the main contributor to functional deficits9, yet few studies have directly investigated regenerative capacity with function.

The possibility to utilize biomarkers to predict tendon healing has been suggested but requires further investigation for validation. After ATR, late surgical intervention leads to an imbalance in tendon homeostasis that is time-dependent10, but has yet to be related with AT function.

Properties of the healed tissue are difficult to characterize in vivo. One available method measures AT length changes using ultrasonography during active muscle function11. The AT’s capacity to store and transfer forces is due to its compliance, which allows muscle efficiency12; following ATR, this capacity is diminished.

This research aims to link the AT’s intrinsic regenerative capacity at the time of surgery with resultant healed AT tissue function. Our goal is to identify markers predictive of human ATR healing outcomes by combining investigations of early healing events with late-stage tissue function.

Tearm: Alison Agres, Kirsten legerlotz, Britt Wildemann, Sebastian Manegold

Funding: Einstein Kickbox - Young Scientists

Einstein Kickbox Projects - Förderperiode 2017

Fighting liver cirrhosis? Establishment and analysis of decellularized human cirrhotic liver slices as a 3-dimensional model to study cell matrix interactions

Liver cirrhosis is one of the main indications for liver transplantation. Due to the organ shortage, this therapy option is limited to the minority of patients suffering from cirrhosis. Therefore, there is a need of alternative treatment options.The aim of our project is to establish a decellularization protocol for human cirrhotic livers slices, which preserves the natural extracellular matrix (ECM) of cirrhotic livers. These decellularized liver slices will serve as a 3 dimensional model to study cell matrix interactions. If we are able to establish a protocol which will preserve the ECM, we will conduct in vitro recellularization experiments to study how the cirrhotic ECM will change the genotype and phenotype of different cell types. With this knowledge we aim to modify specific cell types in vivo or vitro for example prior to cell transplantation. Our ambition is to steer the cell matrix interaction via these modified cells after their transplantation and thereby halt or even reverse the progress of liver cirrhosis. This approach may offer an alternative treatment option in the future.

Team: Karl Hildebrandt, Oliver Klein, Igor Sauer    

Funding Scheme: Kickbox

In need for neuroregeneration ! - Development of a novel targeted drug for stroke treatment beyond the time window of thrombolysis

After stroke, the brain can compensate for lost neuronal tissue functions by rewiring of the neuronal network. However, in adult brains, post-ischemic healing is compromised, since neuroplasticity is limited by the adult shape of ECM consisting of a very dense meshwork of ECM proteins. We have shown, that endothelial cells can steer the post ischemic ECM remodeling depending on the inflammatory Stat3 pathway (Hoffmann et al., 2015). In this project, we want to shape the inflammatory endothelial cell response after stroke using a targeted drug for endothelial Stat3 activation. This will steer the remodeling of the ECM towards a neuroplasticity permissive status. This might help to overcome the compromised healing and regeneration after stroke. We will evaluate the drug in mice using a transient filamentous occlusion of the middle cerebral artery (MCAo) as stroke model and behavioral tests to determine effects on long-term functional outcome. We will monitor regenerative effects by visualization of neuronal network changes by viral and chemical tracing techniques as well as DTI and resting state MRI sequences. ECM remodeling will be determined histologically with an emphasis on the perineuronal net and CNS Ranvier´s node ECM.    

TeamChristian Hoffmann, Christoph Harms, Gisela Lättig, Philipp Böhm-Sturm

Funding Scheme: Kickbox

BMPingECM– BMP/TGFβ regulation within the extracellular matrix: From rare disease cellular models to altered ECM and cell- mechanics

The extracellular matrix (ECM) instructs cells to orchestrate crucial aspects of their biological behavior. As a multidisciplinary team of cell biologists, biochemists and biophysicists we set out to investigate biophysical and biochemical features of the ECM from endothelial cells with different signalling backgrounds and found striking changes in its composition and growth factor reservoir capacity. We uncovered feedback regulations affecting transcriptional responses, adaptations of the cytoskeleton and the ECM and altered mechanical properties explaining the vicious cycle in the development of vascular diseases.

The ECRT support enables us to characterize these biochemical and biophysical characteristics of the altered ECM with unprecedented resolution and to provide new mechanisms that potentially help to understand the involvement of BMPs and TGFb in successful and failing endogenous repair scenarios.

Team: Christian Hiepen, Petra Knaus, Kerstin Blank, Peter Fratzl

Funding Scheme: Kickbox

How do cellular dynamics shape vascular network structure?

Angiogenesis is a fundamental process in development, cancer, and regeneration. Complex signaling pathways regulate tip selection, tip migration, and stalk cell proliferation to shape the emerging vascular network. To improve our basic understanding of how signaling interactions within individual cells determine vascular network architecture, we need multi-scale computational models that integrate the cellular signaling dynamics into tissue level dynamics. In the kickbox phase, we will implement the software necessary to efficiently simulate intracellular dynamics in multi scale agent based models and generate a proof of concept study.In the follow up project, we aim to collaborate with experimental scientists to iteratively generate, test, and refine hypotheses on how intracellular signalling defines vascular network structure.

Team: Clemens Kühn, Judith Wodke, Sara Checa Esteban, Edda Klipp, Georg Duda

Fundind Scheme: Kickbox

Uncover the Role of IL-4 in Hyaline Cartilage Homeostasis and Osteoarthritis

The objective of the project is to uncover the function of IL-4 in cartilage homeostasis with available murine and human material applying different methods. Furthermore, we will work on a concept to implement in silico methods as a strong tool to study biological processes and signalling pathways and to use the power of an interdisciplinary collaboration towards a common goal. In a following (advanced) project, we aim to clarify the role of IL-4 in the pathogenesis of OA, to provide evidence-based approaches on how to overcome these compromised conditions and to identify new therapeutic targets towards OA to challenge an unmet clinical need that has remained unresolved for far too long. 

Team: Annemarie Lang, Katharina Schmidt-Bleek, Max Löhning, Susanna Röblitz, Frank Buttgereit

Funding Scheme: Kickbox

Quantifying the quadricep force in exercise mediated osteoarthritis therapies

Our project is aimed to prototype new mathematical models which explain the forces produced by the quadricep in exercise meditated OA therapies. We wish to extend existing isometric quadricep models to general motions by enriching the equations using prosthesis data. We are looking for groups working in biomechanics and focused on quantifying qualitatively the effects of exercise mediated therapies.

TeamVikram Sunkara, Max Von Kleist, Georg Bergmann

Funding Sceme: Kickbox

The Role of T-cell Activation Levels in Periprosthetic Joint Infection Development

With increasing life expectancy and concurrent high demands regarding personal mobility, the numbers of total joint arthroplasties (TJA) are rising. Accordingly, larger numbers of infections are documented. Periprosthetic Joint Infections (PJIs) account for the majority of implant failure, occurring in 2- 3% of all primary TJAs with even higher rates of 3-10% in revision surgeries. A consistent diagnostic after TJA is critical for the therapeutic success. Particularly the diagnostics of persistent low grade infections is still insufficient, calling for the development of highly sensitive diagnostic tools for early detection.

Immunologically reactive molecules are released from T-cells upon cell activation. We hypothesize that selected molecules can not only be used to assess the level of T-cell activation and the reactivity of the adaptive immune response, but also as a marker for lingering infections. Furthermore, their level may serve to clarify an underlying molecular predisposition for endogenous infection management and potentially even resume a protective role in infections.

Defining a biomarker on protein level, that enables local, minimally invasive and reliable diagnostics in the field of PJI while giving insight into the patients’ general immune status, is an ambitious, novel and innovative approach.

The project elements for the KickBoxSeed-Grant are in the focus area of “cellular mechanisms in compromised healing” while the long-term vision of the project additionally addresses the local modulation of inflammation. The concept provides an interface between cell-based immunology and pathogen-based infectiology/immunology with endoprosthetic infections as a clinically highly relevant field of application.

Team: Andrea Sass, Michael Fuchs, Simon Reinke, Anke Dienelt, Janine Mikutta, Andrej Trampuz, Carsten Perka, Juri Rappsilber

Funding Scheme: Kickbox