Virus Entry von HCV
© Prof. Dr. Gisa Gerold
Einwirkungen auf Patienten
© Dr. Graham Brogden
Flussdiagramm der Projektplanung
© Prof. Dr. Gisa Gerold

Current Projects

Characterization of Hepatitis C Virus Entry and the Influence of Host Genetics on Infection

Fluoreszenzmiskroskopische Aufnahme
Localization of SRFBP1 in Human Hepatoma Cells. SRFBP1 is an HCV Entry Factor Identified by Quantitative Proteomics
© Prof. Gisa Gerold

Hepatitis C virus (HCV) infects 71 million individuals worldwide and can cause fibrosis, cirrhosis and liver cancer. The virus uses the human transmembrane protein CD81 to infect liver cells. Since CD81 lacks signaling domains, we had hypothesized that it coordinates HCV uptake through protein interactions with membrane proximal signaling adaptors and cytoskeleton regulators. Our previous work identified 33 CD81 protein interactions in human hepatoma cells and could show that at least ten of the CD81 protein interactors are required for HCV infection. We now aim at understanding the molecular function of new host factors as well as the impact of human genetic variations in these factors on susceptibility and disease progression. We further analyze if the host factors are differentially used by the seven HCV genotypes and by other RNA viruses. Finally, we ask if the host factors contribute to the narrow tissue and species tropism of HCV. This work will identify entry mechanisms of an important human pathogen and may reveal genetic causes of differential disease progression and therapy success in hepatitis C patients.

Collaboration Partners

Felix Meissner (Max-Planck-Institut für Biochemie, Martinsried),
Lars Kaderali (Universität Greifswald),
Jan Phillip Machtens (EZ Jülich).

Funding

DFG (GE  2145/3-1), DFG (GE  2145/3-2), DFG (SFB900-C7), DZIF (TI 07.003), Deutsche Leberstiftung (S163/10073/2011), HFSPO (LT000048/2009-L), Leopoldina (LPDS 2009-9), Excellence Cluster RESIST

Project-related Publications

Single-nucleotide variants in human CD81 influence hepatitis C virus infection of hepatoma cells.
Alberione MP, Moeller R, Kirui J, Ginkel C, Doepke M, Ströh LJ, Machtens JP, Pietschmann T, Gerold G. Med Microbiol Immunol. 2020 Apr 22:1-16.

Hepatitis C Virus Entry: Protein Interactions and Fusion Determinants Governing Productive Hepatocyte Invasion.
Gerold G, Moeller R, Pietschmann T. Cold Spring Harb Perspect Med. 2020 Feb 3;10(2):a036830.


Bruening J, Lasswitz L, Banse P, Kahl S, Marinach C, Vondran FW, Kaderali L, Silvie O, Pietschmann T, Meissner F*, Gerold G* (2018) Hepatitis C virus enters liver cells using the CD81 receptor complex proteins calpain-5 and CBLB. PLoS Pathog 14(7): e1007111.. *these authors contributed equally

Lasswitz L, Chandra N, Arnberg N*, Gerold G* (2018) Glycomics and Proteomics Approaches to Investigate Early Adenovirus-Host Cell Interactions. J Mol Biol 430(13): 1863-1882. Review*these authors contributed equally

Banse P, Moeller R, Bruening J, Lasswitz L, Kahl S, Khan AG, Marcotrigiano J, Pietschmann T, Gerold G (2018) CD81 Receptor Regions outside the Large Extracellular Loop Determine Hepatitis C Virus Entry into Hepatoma Cells. Viruses 10(4).

Gerold G, Bruening J, Weigel B, Pietschmann T (2017) Protein interactions during the flavivirus and hepacivirus life cycle. Mol Cell Proteomics 16(4 suppl 1): S75-S91.

Gerold G, Meissner F, Bruening J, Welsch K, Perin PM, Baumert TF, Vondran FW, Kaderali L, Marcotrigiano J, Khan AG, Mann M, Rice CM, Pietschmann T (2015) Quantitative Proteomics Identifies Serum Response Factor Binding Protein 1 as a Host Factor for Hepatitis C Virus Entry. Cell Rep 12(5): 864-878.

Emerging Virus Host Factor Discovery

Flussdiagramm der Projektplanung
Quantitative Proteomics to Study Virus Host Cell Interactions.
© Prof. Dr. Gisa Gerold

Mass spectrometry based interaction proteomics has become highly sensitive and quantitative. Using state of the art proteomics, we search for cell surface proteins engaged by human pathogenic viruses. Specifically, we employ label free quantification (LFQ) to define the interactome of known host factors such as the SARS-CoV-2 receptor ACE2 and the promiscuous virus attachment factor TIM-1. In a complementary approach, we use proximity labeling (APEX2) and targeted crosslinking approaches to identify proteins, which interact with viral and host proteins during infection. In follow up experiments, we address the functional role of interaction partners using CRISPR/Cas9 knockout, RNA silencing and blocking techniques.

Our major pathogens of interest are zoonotic, re-emerging, mosquito-borne human pathogens of the alphavirus and phlebovirus genus, including Chikungunya virus and Rift Valley Fever Virus. Identified entry and replication factors will be tested for their specificity to various virus strains and their expression in different human tissues targeted by the virus. Lastly, we will analyze if orthologs in mosquitoes and vertebrates, which serve as transmission and reservoir hosts, also function as host factors. The work will shed light on how zoonotic viruses infect host cells with putative implications for antiviral strategy development.

Collaboration Partners

Margaret Kielian (Albert Einstein College, New York, NY, USA),
Pierre-Yves Lozach (Heidelberg University Hospital, Germany),
Charles M. Rice (Rockefeller University, New York, NY, USA),
Niklas Arnberg (Umea University, Sweden)

Funding

DFG (GE  2145/3-2), German Liver Foundation (S163/10135/2017), DAAD, ZIB, Friends of the MHH, Knut and Alice Wallenberg Foundation, Kempe Foundation

Project-related Publications

Quantitative Proteomics of Uukuniemi Virus-host Cell Interactions Reveals GBF1 as Proviral Host Factor for Phleboviruses.
Uckeley ZM*, Moeller R*, Kühn LI, Nilsson E, Robens C, Lasswitz L, Lindqvist R, Lenman A, Passos V, Voss Y, Sommerauer C, Kampmann M, Goffinet C, Meissner F, Överby AK, Lozach P#Y, Gerold G#. Mol Cell Proteomics. 2019 Dec;18(12):2401-2417. *,#equally contributed

Dynamic Dystroglycan Complexes Mediate Cell Entry of Lassa Virus.
Herrador A, Fedeli C, Radulovic E, Campbell KP, Moreno H, Gerold G, Kunz S. mBio. 2019 Mar 26;10(2):e02869-18

Characterization of the Filovirus-Resistant Cell Line SH-SY5Y Reveals Redundant Role of Cell Surface Entry Factors.
Zapatero-Belinchón FJ, Dietzel E, Dolnik O, Döhner K, Costa R, Hertel B, Veselkova B, Kirui J, Klintworth A, Manns MP, Pöhlmann S, Pietschmann T, Krey T, Ciesek S, Gerold G, Sodeik B, Becker S, von Hahn T. Viruses. 2019 Mar 19;11(3):275.


Fedeli C, Torriani G, Galan-Navarro C, Moraz ML, Moreno H, Gerold G, Kunz S (2018) Axl Can Serve as Entry Factor for Lassa Virus Depending on the Functional Glycosylation of Dystroglycan. J Virol 92(5).

von Schaewen M, Dorner M, Hueging K, Foquet L, Gerges S, Hrebikova G, Heller B, Bitzegeio J, Doerrbecker J, Horwitz JA, Gerold G, Suerbaum S, Rice CM, Meuleman P, Pietschmann T, Ploss A (2016) Expanding the Host Range of Hepatitis C Virus through Viral Adaptation. MBio 7(6)

Scull MA, Shi C, de Jong YP, Gerold G, Ries M, von Schaewen M, Donovan BM, Labitt RN, Horwitz JA, Gaska JM, Hrebikova G, Xiao JW, Flatley B, Fung C, Chiriboga L, Walker CM, Evans DT, Rice CM, Ploss A. (2015) Hepatitis C virus infects rhesus macaque hepatocytes and simianized mice. Hepatology. 62(1):57-67.

Gerold G, Bruening J, Pietschmann T. (2015) Decoding protein networks during virus entry by quantitative proteomics. Virus Research. 218:25-39.

Kapoor A, Simmonds P, Gerold G, Qaisar N, Jain K, Henriquez JA, Firth C, Hirschberg DL, Rice CM, Shields S, Lipkin WI. (2011) Characterization of a canine homolog of hepatitis C virus. PNAS. 108(28):11608-13.

HCV – arena- and bunyavirus co-infection and modulation of innate sensing mechanisms

HCV is a small enveloped RNA virus and the causative agent of hepatitis C. It affects 71 million individuals worldwide and can cause severe liver disease including cirrhosis, fibrosis and hepatocellular carcinoma. The virus is underdiagnosed and thought to be highly prevalent in African regions, where outbreaks of hemorrhagic fever viruses such as the arenavirus Lassa virus and of bunyaviruses occur. Since chronic HCV infection alters the immune status of the liver, which is also a target organ for arena- and bunyaviruses, we hypothesized that a co-infection with HCV and a secondary virus may alter the severity of disease. Using cell culture models of hepatoma cells and primary hepatocytes, we address how a co-infection impacts virus propagation, cellular innate immune responses and sensitivity to licensed antiviral drugs. These efforts will allow the assessment of risks associated with underlying chronic hepatitis during outbreaks with emerging and re-emerging arena- and bunyaviruses.

Collaboration Partners

Stefan Kunz (CHUV, Lausanne, Switzerland),
Friedemann Weber (University of Gießen),
Magnus Evander (Umea University, Sweden),
Clas Ahlm (Umea University, Sweden).

Funding

Deutsche Leberstiftung (S163/10135/2017), DAAD, ZIB, Freunde der MHH, Kempe Foundation, Knut and Alice Wallenberg Foundation

Project-related Publications

Quantitative Proteomics of Uukuniemi Virus-host Cell Interactions Reveals GBF1 as Proviral Host Factor for Phleboviruses.
Uckeley ZM*, Moeller R*, Kühn LI, Nilsson E, Robens C, Lasswitz L, Lindqvist R, Lenman A, Passos V, Voss Y, Sommerauer C, Kampmann M, Goffinet C, Meissner F, Överby AK, Lozach P#Y, Gerold G#. Mol Cell Proteomics. 2019 Dec;18(12):2401-2417. *,#equally contributed


Moreno H, Moller R, Fedeli C, Gerold G, Kunz S (2019) Comparison of the innate immune response to pathogenic and non-pathogenic Clade B New World arenaviruses. J Virol 93(19).

Bruening J, Weigel B, Gerold G. (2017) The role of type III interferons in hepatitis C virus infection and therapy. J Immunol Res. 2017:7232361.

Virus Entry von HCV
Innate immunity to HCV infection and viral evasion mediated by the viral NS3/4A protease (modified from Weigel, Bruening, Gerold, Journal of Immunology Research, 2016).
© Prof. Dr. Gisa Gerold

Paving the Way towards Personalized Prevention and Care of Severe Norovirus Gastroenteritis

Noroviruses are a major cause of gastroenteritis and this leads to a significant economic burden. Acute outbreaks on cruise ships and in elderly care facilities as well as chronic norovirus infections in immunocompromised individuals, such as transplant patients, cause a severe health risk. To date, no vaccine or specific treatment options exist and we have limited knowledge about the inter-individual differences that influence the outcome of a norovirus infection. Determining the parameters that render a person more or less prone to norovirus infection and that determine the severity of infection is therefore important in order to devise strategies to prevent and treat norovirus gastroenteritis. The consortium PRESENt (‘Paving the Way towards Personalized Prevention and Care of Severe Norovirus Gastroenteritis’) brings together scientists from Hannover Medical School, TWINCORE, L3S Research Center and the Helmholtz Institute of Infection Research. All consortia partners are associated with the newly established Centre for Individualised Infection Medicine, CiiM. The goal of the consortium is to investigate the role of individual parameters such as age, gender, gastrointestinal microbiota and the virus associated human biomolecules in norovirus infection. The PRESENt team will evaluate these parameters in a retrospective and prospective clinical study. Furthermore, differences in disinfectant efficacy for a broad range of norovirus patient isolates will be determined using state of the art in vitro infection models (so called organoids). ‘Machine learning’ methods and data intensive technology will identify predictive signatures for severe norovirus infection. The knowledge gained will ultimately guide the development of personalized strategies to individually predict, prevent and treat severe norovirus gastroenteritis.

Project Partners

Prof. Dr. Wolfgang Nejdl (Leibniz Universität Hannover & Technische Universität Braunschweig, Forschungszentrum L3S Hannover),
Prof. Dr. Dr. Michael Marschollek (Medizinische Hochschule Hannover, Peter L. Reichertz Institut für Medizinische Informatik der TU Braunschweig und der Medizinischen Hochschule Hannover),
Prof. Dr. Till Strowig, Prof. Dr. Lothar Jänsch (Helmholtz-Zentrum für Infektionsforschung Braunschweig),
PD Dr. Benjamin Heidrich (Medizinische Hochschule Hannover, Klinik für Gastroenterologie, Hepatologie und Endokrinologie Hannover),
Prof. asoc. inv. Dr. Gisa Gerold (Medizinische Hochschule Hannover, TWINCORE - Zentrum für experimentelle und klinische Infektionsforschung Hannover)

Speaker: Prof. asoc. inv. Dr. Gisa Gerold
Deputy speaker: PD Dr. Benjamin Heidrich

Collaboration Partners

Prof. Dr. Thomas F. Schulz (Medizinische Hochschule Hannover, Institut für Virologie), 
Prof. Dr. Lennart Svensson (Linköping University Schweden), 
Nihal Altan-Bonnet, Ph.D. (National Heart, Lung, and Blood Institute, NIH, Bethesda, USA)

Funding

Niedersächsisches Ministerium für Wissenschaft und Kultur und Volkswagen Stiftung im Rahmen der Ausschreibung "Big Data in den Lebenswissenschaften der Zukunft".

Einwirkungen auf Patienten
Integrated Multimodel Framework to Classify Norovirus Disease Severity.
© Dr. Graham Brogden

Further Projects of the Group