Research

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: 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: 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: 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: 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

Funding: QS-Wissenschaftsfonds, vertreten durch Qualität und Sicherheit GmbH, Bonn, 39.000 EUR

Project Details:
Tierwohl und Tiergesundheit haben in den letzten Jahren einen stetig wachsenden Stellenwert auf allen Stufen der Produktionskette von Nutztieren eingenommen. So wird in vielen Bereichen eine zentrale Tiergesundheitsdatenbank gefordert. Zwar ist in verschiedenen Bereichen der Tiergesundheit eine standardisierte Erfas-sung von Daten gesetzlich vorgeschrieben bzw. durch private Unternehmen abge-deckt, jedoch erfolgt bisher keine zentrale und vor allem keine gesamthafte Aus-wertung, die die Wechselwirkungen der Informationen berücksichtigt.
In diesem Projekt werden Daten von Mastschwein haltenden Mitgliedsbetrieben der Qualität und Sicherheit GmbH aus drei Jahren ausgewertet. Dabei werden verschiedene Gesundheitsvariablen aus den Bereichen Antibiotikaeinsatz, Salmo-nellenmonitoring, Schlachtbefunde und Biosicherheit miteinander verknüpft und sowohl deskriptiv als auch in statistischen Modellen ausgewertet, um strukturelle Zusammenhänge und Wechselwirkungen zu erkennen. Der Erkenntnisgewinn kann als Entscheidungsmatrix zur Identifikation von Betrieben genutzt werden, die bei rein kumulativer Betrachtung einzelner Merkmale nicht auffällig sind, aber den-noch eine spezifische Option zur Verbesserung der Tiergesundheit besitzen (z.B. Betriebe mit geringem Antibiotikaeinsatz bei gleichzeitiger gesundheitlicher Problematik).

Funding: Niedersächsisches Ministerium für Wissenschaft und Kultur, 50.000 EUR

Project Details:
Mit dem Innovation plus-Projekt wird ein Training klinisch-praktischer und kommunikativer Fertigkeiten mit anschließender formativer elektronischer eOSCE-Prüfung (Electronic Objective Structured Clinical Examination) für Studierende, die ihren Zyklus des Praktischen Jahres in der Klinik für Heimtiere, Reptilien, Zier- und Wildvögel der Stiftung Tierärztliche Hochschule Hannover absolvieren, konzipiert und umgesetzt werden. Das Training dient der Vorbereitung der Studierenden auf ihr Praktisches Jahr sowie der tierärztlichen Tätigkeit im Anschluss an das Studium. In dem Zusammenhang werden Modelle und Simulatoren sowie Lernstationen für den Heimtiersektor erstellt für die Vermittlung klinisch-praktischer und kommunikativer Fertigkeiten. Im Anschluss an das praktische Training erfolgt eine Überprüfung der erlernten Fertigkeiten mittels eOSCE-Prüfung.

Funding: BMBF, 114.582 EUR

Project Details:
Obwohl auf Genomebene in Fledertieren eine Vielzahl von Viren nachgewiesen wurden, gibt es nur eine geringe Zahl, die in Form infektiöser Virusisolate vorliegen. Da der Nachweis von Fledertierviren sehr häufig aus Kotproben erfolgt, sollten differenzierte Darmzellen die Zielzellen vieler dieser Viren sein, weshalb sie für die Isolierung von Fledertierviren aus Kotproben besonders geeignet erscheinen.
Ziel des Projektes ist die Etablierung eines Kultursystems bestehend aus intestinalen Organoiden von Fledermäusen, mit dem es ermöglicht werden soll, Fledermausviren zu isolieren und ihre Vermehrungseigenschaften sowie ihr zoonotisches Potential näher zu charakterisieren.

Results:

https://journals.asm.org/doi/10.1128/spectrum.03098-22

Funding: EU Commission, Directorate-General for Health and Food Safety, 724.000 EUR

Project Details:
Finanzielle Zuwendung für die Arbeiten am EU Referenzlabor für Klassische Schweinepest
(Work program according to Annex IV of the Council Directive 2001/89/EC)

Cooperation Partners:

Dr. Christoph Staubach (FLI Riems)