Projects Virus Interaction Proteomics

Characterization of CD81 receptor interactors in hepatitis C virus and Plasmodium liver cell entry

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). 

Enveloped virus entry factor discovery

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).

HCV – arenavirus 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 arena- and bunyavirus outbreaks. 

This work is funded by the German Liver Foundation (S1 63/10135/2017) 

Collaboration partners: Stefan Kunz (CHUV, Lausanne, Switzerland), , Friedemann Weber (Universität Gießen), Magnus Evander (Umea University, Sweden), Clas Ahlm (Umea University, Sweden).


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

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 Hanover Medical School, TWINCORE Centre for Clinical and Experimental Infection Research in Hanover, L3S Research Center in Hanover and Helmholtz Centre for Infection Research in Braunschweig. All consortia partners are associated with the newly established Centre for Individualised Infection Medicine, CiiM, in Hanover. 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.     

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)

Collaborating groups: Prof. Dr. Thomas F. Schulz (Hannover Medical School, Institute for Virology), Prof. Dr. Lennart Svensson (Linköping University Sweden), Nihal Altan-Bonnet, Ph.D. (National Heart, Lung, and Blood Institute, NIH, Bethesda, USA) 




Characterization of CD81 receptor interactors in hepatitis C virus and Plasmodium liver cell entry

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

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


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.

Virus entry factor discovery: Mosquito-borne viruses and enteric viruses

Uckeley ZM*, Moeller R*, Kuhn LI, Nilsson E, Robens C, Lasswitz L, Lindqvist R, Lenman A, Passos V, Voss Y, Sommerauer C, Kampmann M, Goffinet C, Meissner F, Overby AK, Lozach PY#, Gerold G# (2019) Quantitative proteomics of Uukuniemi virus - host cell interactions reveals GBF1 as proviral host factor for phleboviruses. Mol Cell Proteomics 18(12): 2401-2417.*,#equally contributed

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

Zapatero-Belinchon FJ, Dietzel E, Dolnik O, Dohner K, Costa R, Hertel B, Veselkova B, Kirui J, Klintworth A, Manns MP, Pohlmann S, Pietschmann T, Krey T, Ciesek S, Gerold G, Sodeik B, Becker S, von Hahn T (2019) Characterization of the Filovirus-Resistant Cell Line SH-SY5Y Reveals Redundant Role of Cell Surface Entry Factors. Viruses 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. 


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

HCV - arenavirus co-infection and modulation of innate sensing mechanismus

Moreno H, Möller R, Fedeli C, Gerold G, Kunz S (JVI00160-19) Comparison of the innate immune response to pathogenic and non-pathogenic Clade B New World arenaviruses J Virol. 2019 Jul 3. pii: JVI.00148-19. doi: 10.1128/JVI.00148-19. [Epub ahead of print]

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.