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Laboratory of in vitro modeling systems of pulmonary and thrombotic diseases
Welcome to the laboratory of "in vitro modeling systems of pulmonary and thrombotic diseases" which was founded during the corona pandemic in 2020 by Dr. Robert Szulcek following his appointment as Professor of Physiology at the Institute of Physiology at the Charité – Universitätsmedizin Campus Mitte and the German Heart Center Berlin, Germany.
We aim to bridge the translational gap between animals and humans with the help of humanized disease models. Our mission is to design animal-free test platforms and biological in vitro models that allow for the study of patient-derived cells (translatability) in their (patho)physiological micro-environment (cellular plasticity) by considering the cellular and structural vascular build-up and biomechanical forces within different vascular compartments (cellular and structural vascular heterogeneity). Thereby a special focus lies on enabling the recording of continuous data (live-cell recordings) with high spatial and temporal resolution.
Our vision is that these platforms are universally used to improve strategies for diagnosis, to explore pathological mechanisms, to test drugs and treatments, and to guide therapy (point-of-care: therapy monitoring and bedside testing).
We have established international collaborations with specialists in clinical fields like pneumology, cardiology, and surgery; as well as in basic and translational research; and bioinformatics. In these networks we can perform everything from multi-center pre-clinical experimentation (Van Der Feen et al., 2019) to small proof-of-principle clinical trials with quick turnaround times (Botros et al., 2020). Our partners include but are not limited to the German Heart Center Berlin; Free University Berlin; Max Planck Institute for Heart and Lung Research Bad Nauheim; Comprehensive Heart Failure Center Würzburg; Amsterdam University Medical Centers; Leiden University Medical Center, University of Barcelona; and the Brigham and Woman’s as well as the Massachusetts General Hospital in Boston, USA.
Mission
- Animal-free: Design in vitro test platforms and humanized disease models
- Cell plasticity: Study cells in their (patho)physiological micro-environment
- Heterogeneity: Consider cellular and structural vascular build-up
- Translatability: Utilize patient samples
- Time-dependency: Record continuous data with high temporal and spatial resolution
Research Portfolio
Our research portfolio focuses on two main thematic areas. Program 1: The interaction of blood components with vascular cells and their extracellular matrix (ECM). Program 2: Cell plasticity and heterogeneity in control of lung vascular remodeling, thrombosis, and hemostasis. Specifically, projects comprise the therapeutic normalization of platelet function in COVID-19; mechanisms of in situ thrombosis in chronic thromboembolic pulmonary hypertension (CTEPH); pharmacological modulation of endothelial kinase profiles in pulmonary arterial hypertension (PAH); and the study of altered lung ECM in PAH.
Program 1: Interaction of blood components with vascular cells and their ECM.
Program 2: Endothelial plasticity and heterogeneity in control of lung vascular remodeling.
Methodological Approach
Our work builds upon close collaborations with clinical departments, the use of patient material, and the isolation and culture of a variety of primary vascular and blood circulating cells (Table 1). We use different state-of-the-art optical and electrical real-time measurements, and multi-omics approaches to characterize these biological samples (Figure 1). The latter includes single-cell and global RNA-sequencing, as well as proteome, and kinome profiling. The generated insights are used to model hallmarks of disease (e.g., extracellular matrix composition, fluid flow profiles) and thereby uncover disease-specific cellular dysfunctions. In general, our methodological and technological approaches cover a wide spectrum of biological experimentation with a methodological focus on in vitro thrombosis modelling (Manz et al., 2020), application of fluid flow (Szulcek et al., 2016), and real-time electric characterization of cell responses to wounding, mechanical and chemical stimuli (Szulcek et al., 2014). These model systems allow exploring pathomechanisms ex-vivo, to study the interaction of vascular compartments and cell types in vitro, and to test new therapeutic strategies (Manz et al., 2022).
Workflow
- Collection of biological material and isolation of patient-derived cells; considering vascular compartments
- Characterization of biological samples by modern “omics” and real-time approaches
- In vitro modelling of biomechanical aspects of pathology to unmask dysfunction
- Exploration of mechanisms of dysfunction and test compounds to restore physiological function under these microenvironments
- Translational focus on pulmonary and thrombotic diseases
Equipment & Facilities
Cell culture facilities
Wet lab facilities
EVOS M5000 fluorescence imaging system
Ibidi climate chamber for live cell imaging and generation of hypoxic conditions
Ibidi and syringe pumps for generation of fluid shear stress
Electric Cell-substrate Impedance Sensing (ECIS) System
Access to all core-facilities of Charité and Berlin Institute of Health (BIH)
Key-publications and Editorial Work
- Research Topic in Frontiers of Physiology - Bridging Techniques: Basic Science of Molecules, Cellular Systems, and Whole-Organ Physiology https://www.frontiersin.org/research-topics/16429/bridging-techniques-basic-science-of-molecules-cellular-systems-and-whole-organ-physiology#articles
- Epigenetic Modification of the VWF Promotor Drives Platelet Aggregation on the Pulmonary Endothelium in Chronic Thromboembolic Pulmonary Hypertension. Manz XD, Szulcek R, Pan X, Symersky P, Dickhoff C, Majolée J, Kremer V, Michielon E, Jordanova ES, Radonic T, Bijnsdorp IV, Piersma SR, Pham TV, Jimenez CR, Vonk Noordegraaf A, de Man FS, Boon RA, Voorberg J, Hordijk PL, Aman J & Bogaard HJ (2022). Am J Respir Crit Care Med. [IF2020= 21.405]
- Methods Collection in JOVE - In Vitro Modeling Systems of Respiratory Illnesses and Diseases www.jove.com/de/methods-collections/343
- Exacerbated Inflammatory Signaling Underlies Aberrant Response to BMP9 in Pulmonary Arterial Hypertension Lung Endothelial Cells. Szulcek R, Sanchez-Duffhues G, Rol N, Pan X, Tsonaka R, Dickhoff C, Yung LM, Manz XD, Kurakula K, Kiełbasa SM, Mei H, Timens W, Yu PB, Bogaard HJ & Goumans MJ (2020). Angiogenesis 23, 699–714. [IF2020=9.596]
- Delayed Microvascular Shear Adaptation in Pulmonary Arterial Hypertension. Role of Platelet Endothelial Cell Adhesion Molecule-1 Cleavage. Szulcek R, Happé CM, Rol N, Fontijn RD, Dickhoff C, Hartemink KJ, Grünberg K, Tu L, Timens W, Nossent GD, Paul MA, Leyen TA, Horrevoets AJ, Man FS de, Guignabert C, Yu PB, Vonk-Noordegraaf A, van Nieuw Amerongen GP & Bogaard HJ (2016). Am J Respir Crit Care Med 193, 1410–1420. [IF2016=13.204]
- Szulcek R, Bogaard HJ & van Nieuw Amerongen GP (2014). Electric cell-substrate impedance sensing for the quantification of endothelial proliferation, barrier function, and motility, J Vis Exp. [IF2014=1.113]
Temporal Staff and Alumni
- Milena Droll - Student intern
- Daniel Steiert - Student assistant Bioinformatics & master thesis Bioinformatics
- Claudia Abad Baucells - Master internship Molecular Medicine
- Lena Kampen - Master internship Molecular Medicine
- Felix Aulenbach - Master thesis Bioinformatics
- Xue D. Manz - PhD student