Research

Funding: SprinD GmbH, 263.866 EUR

Project Details:
The viral genome of an RNA virus is released into the cell, where the viral genome is amplified and its transcripts translated into proteins to form more copies the virus. CRISPR/Cas13 enzymes cleave different sites of the viral genome and its transcripts through a combination of crRNAs. Proliferation of the virus is blocked and transcripts to be translated into viral proteins are reduced.

Cooperation Partners:

Universitätsmedizin Göttingen (UMG), Prof. Elisabeth Zeisberg

Funding: MWK über UMG Göttingen, 44.446 EUR

Project Details:
SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus Type 2) is the causative agent of COVID-19. The course of the COVID-19 pandemic so far has emphatically shown that a coordinated bundling of interdisciplinary and complementary expertise is necessary in order to decode the diverse aspects of the biology, pathology and epidemiology of SARS-CoV-2 and to use the knowledge gained clinically for the treatment of patients as well as for modeling the course of infection in the population. In order to be able to provide and implement such a holistic approach, the federal state of Lower Saxony offers ideal conditions with its internationally renowned science locations. We are therefore applying for financial support for collaborative research to set up a COVID-19 research network in the state of Lower Saxony (COFONI).

Funding: Leibniz-Informationszentrum, Technik und Naturwissenschaften, Technische Universitätsbibliothek (TIB), Hannover, 3.560 EUR

Project Details:
In order to enable the students to think outside the box, the already existing offer at the TiHo is to
Instructional/learning videos are expanded. Film sequences are planned that contain basic information on the aquatic and terrestrial animal species researched at ITAW, with learning units on their anatomical and physiological aspects
Particularities. Film sequences on autopsies of the respective animal species are intended to further deepen the learning content and illustrate the fields of activity of the veterinarians at the ITAW. The approach is interdisciplinary
and can offer veterinary medicine and biology students as well as other interested parties insights into anatomical, physiological and pathological contexts. The basics are available and can be processed by a technically experienced assistant into an online module lasting about 90 minutes.

Funding: MWK Niedersachsen, 69.925 EUR

Project Details:
Since the first description of SARS-CoV2 and the related disease COVID-19 in December 2019 various in vitro and in vivo models were utilized to investigate viral life cycle and pathophysiology as basis for effective treatment strategies. In vivo models used to investigate SARS-CoV2 or respiratory viral infections in general range from small animals models to non-human primate models (Munoz-Fontela et al., 2020). But next to the ethical questions accompanying animal models in scientific research also not all questions related to human respiratory epithelium can be examined with (small) animal models. Immortalized human cell lines like Caco-2, Calu-3 or HEK293T which are regularly used to investigate respiratory viruses like influenza or Human Respiratory Syncytial virus (RSV) and also Corona Virus infections (SARS, MERS) are frequently applied to investigate infection and replication of SARS-CoV2; however, although these cell lines are comparatively easy and cost effective to grow, they only poorly recapitulate physiological conditions of the respiratory epithelium. Even cell lines such as A549 cells barely reflect the phenotype of lung epithelium. Human primary bronchial epithelial cells (hBECs) represent a more physiological in vitro model and are also utilized in SARS-CoV2 research (Hoffmann et al., 2020). These cells can be isolated from donor lung tissue, expanded in vitro and matured in Air-Liquid-Interface (ALI) cultures (Hoffmann et al., 2020). While the main focus of our group is scalable generation and utilization of cardiovascular and respiratory derivatives from induced pluripotent stem cells for disease modelling, drug screening and cellular therapies (Merkert et al., 2019; Katsirntaki et al., 2015; Kempf et al., 2014; Schmeckebier et al., 2013; Zweigert et al., 2011), we have recently also established isolation of primary human airway cells of consistent high quality: Hannover Medical School is the leading German lung transplantation center with about 100 lung transplantation per year. In close collaboration with our transplant surgeons and the group of Prof. Danny Jonigk (Pathology, MHH), we are routinely preparing hBECs from bronchial tissue that can be culture-expanded and cryopreserved for later applications. Also maturation of these hBECs is already well established in our group and matured proximal epithelial cells are already provided to partners for investigation of SARS-CoV2 as well as other respiratory viral infections (e.g. RSV). While these primary hBECs represent a superior in vitro model for SARS-CoV2 infection studies and COVID disease modelling, they do represent only one respiratory compartment. Infection by SARS-CoV2 not only occurs in the upper throat and nasal tract, trachea and bronchi, but also in the alveolar epithelium. Therefore also organotypic in vitro models that reflect the distal lung compartment are urgently required. While maintenance and expansion of isolated type II alveolar epithelial cells (AT2 cells) was so far impossible, substantial progress was very recently made, and protocols that enable production of such cells in larger numbers have been developed. So called Alveolospheres, organoids consisting of alveolar type 1 and 2 cells, allow expansion and maturation of isolated distal epithelial cell in vitro and were recently used to model SARS-CoV2 infection (Karsura et al., 2020; Salahudeen et al., 2020). Although organoid cultures enabled in vitro culture of AT1 and AT2 cells, we believe ALI cultures would represent a more physiological system with air exposure of the epithelial cells and could subsequently allow co-culture systems with endothelial cells as well as macrophages to create more complex organotypic in vitro models of the alveolar lung compartment. By utilizing our experience from differentiation of induced pluripotent stem cells (Katsirntaki et al., 2015; Schmeckebier et al., 2013) we aim to establish isolation, 2D expansion and cryopreservation of human AT2 cells, and further maturation and differentiation in type I alveolar epithelial (AT1) cells in an ALI culture system for in vitro modelling of SARS-CoV-2 infections.

Cooperation Partners:

Prof. Dr. Ulrich Martin, Dr. rer. nat. Ruth Olmer

Leibniz-Forschungslaboratorien für Biotechnologie und künstliche Organe, Klinik für Herz-, Thorax-, Transplantations- und Gefäßchirurgie - Medizinische Hochschule Hannover

Funding: Deutsche Forschungsgemeinschaft (DFG), Bonn plus ein Doktorandenstipendium (HGNI, 1 Jahr, je 1300 ?), 75.850 EUR

Project Details:
CoVid-19 is characterized by infection of the airways by SARS-CoV-2. Apart from the respiratory tract, other organs are also involved, e.g. the intestinal tract. The importance of the intestinal infection is increasingly recognized. In a large proportion of pediatric patients, virus was detected in rectal swabs and virus shedding from the intestine was found even when oral swabs had become negative. Therefore, prolonged virus shedding and fecal-oral transmission have to be considered. This notion is supported by detection of the virus in wastewater.
The aim of this short project is to apply intestinal cell cultures to characterize the infection of differentiated intestinal epithelial cells by SARS-CoV-2 and thereafter intestinal orgranoids.
This in vitro infection approach will target the following aims:
1. Characterization of the replication efficiency of SARS-CoV-2 (virus yield, virus exit, virus entry, apical, basolateral).
2. Localization of the cellular receptor(s) in human intestinal Caco-2 cells
3. Investigation of the trafficking of the cellular receptor(s), determination and subsequent modulation of their mode of interaction with membrane microdomains (lipid rafts, LRs)
4. Effects of glycosylation modulators on the spike glycoprotein and its interaction with intestinal cells
5. Implication of virus infection on the trafficking and function of crucial enzymes of the intestinal physiology (APN, SI, LPH, DPP4).
This project will provide substantial information on the replication of SARS-CoV-2 in intestinal epithelial cells, evaluate its effects on the intestinal function and provide solid hypotheses on the molecular and biochemical basis for the symptoms elicited by SARS-CoV-2 infections. These hypotheses can be then examined at a later stage in intestinal organoids. Further, unravelling the biosynthetic pathway, glycosylation pattern and mode of interaction of the SARS-CoV-2 receptors and its modulation could constitute exquisite targets for potential therapy.

Funding: This work was supported by COVID-19 Research Network of the State of Lower Saxony (COFONI) with funding from the Ministry of Science and Culture of Lower Saxony, Germany (14-76403-184) and by PRACTIS?Clinician Scientist Program of Hannover Medical School, funded by the German Research Foundation (DFG-ME3696/3-1)., 50.000 EUR

Project Details:
Vaccination is an important measure in controlling the COVID-19 pandemic. However, there were pharmocovigilance concerns as a very rare but potentially disastrous complication was identified after application with the ChAdOx1 nCoV-19 vaccine, i.e. cerebral sinus thrombosis, but also further thrombotic complications including ischemic stroke. Meanwhile, the most likely mechanism has been identified as platelet factor 4 (PF4) antibody mediated immune thrombotic thrombocytopenia, a mechanism which is already known from heparin-induced thrombocytopenia (HIT). Interestingly NETs were suggested to be critical for thrombus formation in the HIT-pathophysiology as well. Markers of NETosis were observed in platelet-rich thrombi in the lungs of mice with HIT. Therefore, NETs might also be involved in vaccine-induced immune thrombotic thrombocytopenia (VITT) mediated thrombotic complications and might therefore be a common trunk in the pathogenesis of COVID-19 and vaccine-induced complications. Moreover, biomarkers reflecting NETs burden in the circulation and within tissue specimens (e.g. cerebral thrombi) may be suitable as diagnostic, therapeutic or prognostic targets in thrombotic diseases. Most importantly, there is a tremendous need to identify patients at risk for cerebrovascular complications due to both COVID-19 and VITT. The proposed project ISI-VITT is intended to contribute to addressing this issue.

Cooperation Partners:

Cerebrovascular Research Group (Head: Prof. Dr. Karin Weissenborn), Deptartment of Neurology, Medical School Hannover: Dr. Ramona Schuppner & Dr. Gerrit M. Grosse

Prof. Dr. Christine Falk, Institute for Transplantation Immunology, Medical School Hannover

Funding: SprinD, 684.523 EUR

Project Details:
Current antiviral agents mainly target stages in the viral life cycle such as viral attachment to host cell or replication of viral RNA and DNA. Most of the available antiviral agents are effective against replicating viruses only, and due to the lack of specificity, many have adverse side effects. Particularly in endemic and pandemic disease outbreaks, there is additional challenge through virus mutagenesis and the development of viral variants. Therefore, approaches, which target various virus variants are urgently needed. Current paradigms in antiviral treatment involve usage of small molecules and/or therapeutic antibodies. Small molecules often have secondary targets and thus can cause side effects. Antibodies are expensive, their administration is mostly limited to clinical settings, and also they are affected by mutations. In an endemic or pandemic situation, therapies which have broad coverage within virus families, prevent transmission, and can be safely applied during mild and moderate illness are especially valuable.

Our novel approach aims to fill this need through the use of CRISPR/Cas13, an enzyme from bacteria, which cleaves RNA, including the viral genome (of RNA viruses) and of viral mRNA, thereby blocking viral replication and generation of viral proteins. Our therapeutic strategy has no secondary targets, and can be produced at GMP level at low cost. Through a specific combination of so-called crRNAs, Cas13 is directed to different viral mRNAs and to different sites within the viral genome. The crRNAs are selected such that no human RNA is targeted, and therefore no adverse side effects can be expected. This technology can easily be adapted for any single stranded RNA virus. Through targeting different and highly conserved regions, this approach antagonizes also emerging variants of the original virus.

Cooperation Partners:

Prof. Dr. Elisabeth Zeisberg, Universitätsmedizin Göttingen

Funding: Niedersächsischen Ministeriums für Wissenschaft und Kultur , 3.452 EUR

Project Details:
The aim was to create a course on the topic of "Basic training in the use of fluorescence microscopy" for students of biology and veterinary medicine via the documentation system in the TiHo Moodle platform. This module can be completed by the students in self-study. It is underpinned by texts, videos and subject-specific questions that must be answered after processing and viewing the videos. Essential parts of the content are also made available online as summarizing units as OER in the Twillo portal and are particularly suitable for students from natural science courses, pupils and trainees in (veterinary) medical professions.

Funding: COVID 19 Forschungsnetzwerk Niedersachsen COFONI, 48.638 EUR

Project Details:
Early detection of individuals with a SARS-CoV-2 infection remains the most important tool to interrupt the chain of infection. Several studies have already proven canines? extraordinary olfactory acuity to detect SARS-CoV-2-infected individuals. However, with increasing knowledge about the virus, the variety and chronicity of the disease expression and an increase in the number of people that are vaccinated, some questions have not yet been answered. We will test a variety of hypotheses to further explore the use of detection dogs as a screening method for SARS-CoV-2-infections.

Results:

Dogs can distinguish SARS-CoV-2-infections from other viral infections. However, compared to earlier studies our scent dogs achieved lower diagnostic sensitivities. To deploy COVID-19 detection dogs as a reliable screening method it is therefore mandatory to include a variety of samples from different viral respiratory tract infections in dog training to ensure a successful discrimination process.

https://doi.org/10.3389/fmed.2021.749588

Cooperation Partners:

Medizinische Hochschule Hannover

Funding: Niedersächsische Ministerium für Wissenschaft und Kultur, 1.225.000 EUR

Project Details:
Auf das Coronavirus trainierte Spürhunde werden in diesem Herbst bei mehreren Musikveranstaltungen eingesetzt und mit Antigen- und rtRT-PCR-Tests verglichen. Dies wird weitere Erkenntnisse darüber liefern, wie Corona-Spürhunde am besten bei der Detektion infizierten Menschen eingesetzt werden können.

Results:

www.backtoculture.de

Cooperation Partners:

Medizinischen Hochschule Hannover,

Universitätsklinikum Hamburg-Eppendorf,

Hannover Concerts,

Proevent,

Awias Aviation Services