Ex vivo technologies
Genetic engineering is the manipulation of an organism’s genes using biotech methodologies and it includes several applications that are widely adopted to study cardiac biology. In the past years, important advances have been achieved in the field (viral vector-mediated gene transfer, CRISPR cas9 gene editing etc) making these technologies rapid, efficient and more affordable. These methods are now used to answer specific research questions and they are applied to both in vitro and in vivo research models. The group is testing these novel technologies in a regenerative medicine perspective.
Working closely with the other research groups within Prof Thomas Thum's laboratory, and their great expertise in non-coding RNAs (ncRNAs), the group research interest is mainly focused on the role of ncRNAS in regeneration and the evaluation of both structural and functional parameters which are affected by the therapeutic treatments. The group uses several in vitro and ex vivo models, with a particular focus on living myocardial slice. In a translational perspective, human myocardial slices are also extensively used. The multicellular environment and preserved functionality during in vitro culture make living myocardial slices the ideal model to develop and test new therapeutic strategies.
Ongoing research projects
HCM is the most frequent inherited cardiomyopathy with a prevalance of at least 1:500 in population. Thickening of left ventricular wall and interventricular septum and highly disordered cardiomyocytes (CMs) and myofibrils, cellular hypertrophy and interstitial fibrosis are hallmarks of HCM. Patients develop symptoms like angina pectoris, dyspnea and life threatening cardiac arrhythmias, leading to sudden cardiac death or heart failure. Most patients with a confirmed mutation carry mutations in genes encoding sarcomeric proteins in β-myosin heavy chain (MYH7, β-MyHC), cardiac myosin-binding protein C (MYBPC3, cMyBPC), cardiac troponins I and T (TNNI3, cTnI, and TNNT2, cTnT).
In our group we use different HCM-associated disease models like animal models, induced pluripotent stem cell-derived cardiomyocytes with HCM-associated mutations and living myocardial slices generated from HCM-patient´s tissue. Using these models we aim to elucidate changes of genetic network and non-coding RNA expressions in HCM which are linked to cardiomyocyte dysfunction, hypertrophy and fibrosis. We aim to develop novel therapeutic strategies to prevent HCM initiation and progression.
Unlike traditional therapeutic strategies for heart failure, regenerative cardiology aims to repair myocardial damage, and particularly to generate new cardiomyocytes to compensate cardiomyocyte loss. This approach aims to regenerate cardiomyocytes in situ through activation of cardiomyocyte proliferation. Recent studies demonstrated that neonatal mammals possess a remarkable regeneration potential that allows them to recover after severe cardiac tissue damage. This capacity is lost during the first week of life, however, it suggests the presence of regenerative circuits, which could be reactivated and used to induce or enhance cardiomyocyte proliferation. MicroRNAs (miRNAs) and non-coding RNA (ncRNA) have potent effects on gene expression and can alter cell phenotype by co-ordinately targeting multiple components in important cellular pathways. This project aims at identifying non-coding RNA networks that are critically involved in cardiomyocyte proliferation and cardiac regeneration, and target them to device novel therapeutic strategies able to boost cardiac regeneration and enhance tissue repair.
Investigation of resident cardiac macrophages in living ventricular myocardial slices and their manipulation using novel molecular tools.
Testing of novel therapeutics against cardiovascular diseases in LMS.
Current group members
- Natalie Weber, MD, PhD, Group Leader
- Naisam Abbas, medical doctor, PhD student
- Marida Sansonetti, PhD student
- Marco Bentele, PhD student
- Florian Johann Gustav Waleczek, medical student, research associate
- Jonas Haas, medical student, research associate
- Jan Weusthoff, medical student, research associate
- Sören Volkerey, medical student, StrucMed
- Noah Richard Scheller, medical student, StrucMed
- Kaja Solveig Menge, medical student, StrucMed
- Angelika Pfanne, MTLA
- Myocardial slice preparation and ex vivo culture (mouse, rat, pig, human)
- Minituarization of LMS for high throughput approaches
- Isometric force measurement in LMS
- Calcium and voltage imaging (Coop Prof. C. Wahl-Schott, Prof. I. Gruh)
- Optimization of AAV-infection, transfection and treatments with substances in LMS
- Biochemical assays (metabolic activity, live/dead staining, expressions of genes and proteins involved in contraction, calcium handling and ion channel activity)
- Histology and morphology analysis of heart tissue
- Retrograde Langendorff-heart perfusion and primary cardiomyocyte isolation
- Primary cardiomyocyte culture
- Measurements of sarcomere shortening and intracellular calcium transients (IonOptix, Coop Prof. T. Kraft, Dr. M. Ricke-Hoch)
- hiPSC-CM differentiation and culture (Coop Dr. R. Zweigerdt)
- Contraction analysis with SarcTrack Software (Coop Dr. C. Toepfer, Dr. Y. Psaras; Prof. E. Ponimaskin, Dr. A. Zeug)
- Calcium imaging
- Ex vivo imaging (Immunofluorescence, live cell imaging)
- Prof. Dr. L. Carrier (Institute for Experimental Pharmacology and Toxicology, UKE, Hamburg)
- Prof. Dr. T. Kraft (Insitute of Molecular- and Cell Physiology, MHH)
- Prof. Dr. C. Wahl-Schott, Prof. Dr. Ponimaskin, Dr. A. Zeug (Insitute of Neurophysiology, MHH)
- Prof. Dr. J. Bauersachs, Prof. Dr. K. C. Wollert, Dr. M. Ricke-Hoch (Department of Cardiology and Angiology, MHH)
- Dr. R. Zweigerdt, Prof. U. Martin (LEBAO, MHH)
- Dr. A. Koroleva, Prof. B. Chichkov (Institute of Quantum Optics, Leibniz Universität, Hannover)
- Dr. C. Toepfer, Dr. Y. Psaras (Radcliffe Department of Medicine, University of Oxford)
- Dr. E. Castillo, Prof. Dr. Beth Pruit (Center for Bioengineering, University of California, Santa Barbara)
We have available projects for StrucMed students, bachelor and master students as well as for FWJ students. Please contact us for further information
Natalie Weber, Weber.Natalie@mh-hannover.de; Phone: +49 511 532-5276