eRapport

Autoimmune Thyroid Diseases: Immuogenetic loci and use of new methods for detection of virus in thyroid tissue with impact on novel therapy methods.

Prosjekt
Prosjektnummer
2014078
Ansvarlig person
Knut Dahl-Jørgensen
Institusjon
Oslo universitetssykehus HF
Prosjektkategori
Postdoktorstipend
Helsekategori
Inflammatory and Immune System, Disputed aetiology and other
Forskningsaktivitet
2. Aetiology
Rapporter
2017
In an in-vitro study we have tested the effect of Hepatitis C virus E2 Protein, and HCV infection in thyrocytes. Our results shows that thyroid cells express HCV receptors; and HCV infection induce upregulation of pathways involved in immune response. We are currently planning for a RTC to test the effect of antiviral treatment in thyroiditis.Our working hypothesis is that viral infection may induce thyroid autoimmunity. In the first part of the study (in cooperation with Prof. Tomer, NY) HCV infection in thyrocytes and islets was used as a model for viral response. A) HCV E2 protein induced increased expression of IL-8, IL-6 and TNFα in in human primary thyrocytes. Using Illumina, RNAseq was performed. Thyrocytes treated with E2 showed increased expression of genes involved in IL-8 pathways both in thyrocytes and hepatocytes. However, in thyrocytes, E2 protein induced genes involved in IFNα pathways and increased expression of several heat shocks that have been reported to be associated with thyroid autoimmunity. The result of this study was published in JCEM, and was reported in American Association Meeting 2017 as the most important discovery in thyroid autoimmunity during last year. B) HCV infection of thyrocytes induced increased expression of pro-inflammatory cytokines, IL-8, TNF α, and IL-6 in thyrocytes. Interestingly, the results in thyrocytes differed with those in hepatocytes. While Infection induced modification in only a minor number of genes, mainly genes involved in apoptotic pathways in hepatocytes, infection induced activation of immune response in thyrocytes. The results of this paper will soon be submitted to JCEM for publication. Our results showed that infection induced upregulation of expression of mir-122 (unpublished data). This may be of the major reason for the high prevalence of thyroid autoimmunity in patients with chronic hepatitis C. Moreover, new therapies such as antimir-122 (already developed and under testing in HCV infection) can be a potential treatment for autoimmunity in this group of patients. C) HCV infection of islets: In HCV-infected islets, TNFα and IL-6 levels increased after infection. Interferon and an HCV polymerase inhibitor reduced viral replication in islet cells. Overall, the expression of miR-122 was low in islets but increased after viral infection. Based on our findings a) detection of a persistent low grade enterovirus infection and/or b) increased expression of viral footprint in newly diagnosed Graves’ disease and Hashimoto thyroiditis and, c) the results in our cell study; we are planning for a randomized clinical trial with antiviral treatment vs. placebo. The protocol for this study is ready. We are currently negotiation the delivery of the study medication/placebo. Patients with newly diagnosed Graves’ disease or Hashimoto thyroiditis will be included. Our primary endpoint will be the number of patients with normal thyroid function at 12 months. Secondary Endpoints: mean thyroid hormone secretion at 3,6,12,24 and 36 months; and the levels of TRAb, TPO-Ab and Tg-Ab at 3, 6, 12, 24 and 36 months. We are continuing the search for virus and viral footprint in already collected thyroid tissue in patients with Graves’ disease and Hashimoto thyroiditis. In collaboration Exeter Medical School (Dr. Sarah Richardson, and Prof. Noel Morgan), we are currently working on detection of EV capsid protein Furthermore, we are analyzing the expression of HLA class I, STATA 1, and PKR. Increased expression of these markers shown to be associated with IFNα expression, possibly in response to viral infection. Furthermore, we will use newest Methods for search for viral RNA (in collaboration with prof. Antonio Toniolo; Varese) in already collected material.

Dette prosjektet er i samarbeid med vårt internasjonal partner professor Yaron Tomer og hans forskings lab i New York, USA. I forbindesle med siste artikkelen som er under arbeid og vil bli snarlig sendt til publikasjon har Sara Hammerstad hatt ett opphold i NY.

2016
Stimulation of human thyroid cells with virus protein or infection of cells with virus induces activation of pathways that may induce autoimmunity by bystander mechanisms. Antiviral treatment will be used in RTC to test the hypothesis that viral infection may be an important trigger and driver for thyroid autoimmunity.In cooperation with Professor Tomer at Mount Sinai Hospital NY-USA, we have conducted cell studies to look for the cellular response to virus protein and virus infection in human thyroid cells. Studies have shown that different viral proteins may individually induce host responses to infection. Hepatitis C virus (HCV) infection was used as a model as thyroiditis is one of the most common extra hepatic manifestations of HCV. We demonstrated that thyroid cells express HCV receptors. Stimulation of human thyroid cells in culture with HCV envelop protein activates a cascade of inflammatory responses. HCV envelope protein induced production of several major cytokines and chemokines involved in autoimmunity. Furthermore, HCV envelop protein induced production of several heat shock proteins. We complemented our study with direct HCV infection of humane thyrocytes and to compare the molecular mechanisms of HCV infection in thyroid with hepatic cells. While HCV infection induces activation of mainly apoptotic pathways, infection of thyrocytes induces activation of by far more genes and cellular pathways. Our data support a key role for viral infection in triggering thyroid autoimmunity by bystander mechanisms. We have further developed a protocol for an intervention study (RTC). We want to test if antiviral treatment may reverse the development of thyroiditis in preclinical conditions, and or shorten then duration of the disease at the onset of disease. We will continue our investigation on viruses and molecular pathways in search for development of new therapy strategies.
2015
Virusinfeksjon kan være en utløsende faktor for utvikling av thyreoidea autoimmunitet. Stimulering av normale humane thyreoidea celler med virus protein eller infisering av cellene induserer produksjon av sentrale cytokiner som er involvert i utvikling av autoimmunitet.Hovedformål med denne studien er å undersøke hvordan virus infeksjon kan bidra til utvikling av thyreoidea autoimmunitet. Årsak til autoimmune thyreoidea sykdommer er ikke helt kartlagt men det er økende evidens at virus infeksjon kan være utløsende faktor i genetisk disponerte personer. Vår tidligere studie viste at virus eller virus markører som interferon, var hyppigere til stede i vevsprøven tatt fra thyreoidea hos pasienter med autoimmune thyreoidea sykdommer (Graves’ hyperthyreose og Hashimoto thyreoiditt) enn i friske personer. I denne studien og i samarbeid med professor Yaron Tomer ved Mount Sina Hospital i NY og Professor Jason Blackard ved Univesity of Cincinnati, har vi undersøkt humane thyreoidea celler og om disse kan bli infisert med virus, og hvilke immunologisk respons som oppstår etter infeksjonen. I dette prosjektet har vi brukt hepatitt C virus (HCV) som en modell for virus infeksjon. Grunnen er at hepatitt C infeksjon ofte har vært assosiert med autoimmune tilstander. Autoimmune thyreoidea sykdommer har vært en av de hyppigste former for ekstrahepatiske manifestasjoner ved Hepatitt C. Hepatitt C virus er et RNA virus som koder for 10 forskjellige proteiner. Studier har vist at de forskjellige virus proteiner alene kan indusere immunologiske reaksjoner. I den første delen av studien har vi påvist at thyreoideaceller på lik linje med hepatocytter uttrykker den hoved reseptor for HCV, CD-81. Stimulering med HCV E2 protein (envelope protein 2) induserer overekspresjon av de fleste HCV reseptorene i thyreoideaceller inkludert CD81. Videre E2 protein induserer en sterk økning i viktige cytokiner og potente kjemokiner i normale humane thyreoidea celler . Disse markørene er kjent for å være involvert i utvikling av autoimmunitet. RNA sekvensering med de nyeste metodene med transkriptom analyse viser oppregulering av flere tusen gener. Mange av disse genene er dokumentert medvirkende i utvikling av autoimmunitet. Ved hjelp av Ingenuity Pathway program identifiserer man mange oppregulert mange pathways. Av særlig interesse er at det ble konstatert en overekspresjon av flere «heat shock proteiner» (HSP). HSP har vist å være involvert i utvikling av autoimmune thyreoidea sykdommer. I den andre delen av studien har vi vurdert effekten av HCV infeksjon på thyreoidea celler sammenlignet med hepatocytter. HCV induserer apoptose i hepatocytter mens thyreoideaceller lettere overlever infeksjonen. Derimot setter infeksjon i thyreoideaceller i gang en rekke immunologiske responser. Resultatene fra RNA sekvensering er under arbeid.
2014
The cause of AITD is not clear but both susceptibility genes and environment factors play an important role. In this study we look for genetic factors and the effect of viral infection in development of thyroid autoimmunity.Autoimmune thyroid diseases (AITD) are prevalent and complex diseases affecting up to 6% of the population. AITD mainly include Graves' disease (autoimmune hyperthyroidism), and Hashimoto thyroiditis (autoimmune hypothyroidism). Studies have shown that GD and HT share some common. AITDs are multifactorial diseases and despite decades of basic research the cause of the disease is still unclear. However, it is of general acceptance that the disease is caused by interplay between genetic factors and environmental triggers. Epidemiological studies, twin studies and family studies and other genetic studies support the importance of environmental factors. In this study we focus on both genetic factors and an important environmental factor, viral infection. In this international project, we are working with highly qualified group with many years of research within genetics, immunology and virology. We have been a part of a study looking for the susceptibility genes in patients with young-age of onset Graves' disease. HLA haplotype has so far shown the strongest association with AITD in all previous studies. The results of our study revealed new susceptibility genes such as BTNL2, NOTCH4, TNFAIP3, and CXCR4, in addition to the well known HLA class II loci. Not surprising, pathway analysis shows the importance of the involvement of immunological cells T helper cells, B cells and antigen presenting cells. The results of this study are published in highly ranked journal JCEM. Lately, it has become clear that the biology and the mechanisms behind diseases are important factors for development of new therapeutic regiments. In collaboration with Dr. Tomer's research lab at Mount Sinai Hospital in NY (Postdoc candidate Sara Hammerstad), we are currently working on the effect of viral infection on thyroid tissue. It is known that 40% of patients with hepatitis C infection present thyroid antibodies, also up to 40% of these patients develop thyroid diseases after treatment with IFNa. In the current study we are studying the mechanisms that viral infection induces autoimmunity. Specifically, we are looking for the effect of hepatitis C virus infection in thyroid cell lines in terms of expression and secretion of cytokines, involved immunological pathways, and epigenetic changes. The results of this study will be presented and published during 2015. Furthermore, we aim to start an intervention study to evaluate the effect of anti viral treatment. We are currently working on development of protocols. If viruses are important for the development of autoimmune thyroid disease, new therapeutic regiments might be developed.
Vitenskapelige artikler
Blackard JT, Kong L, Lombardi A, Homann D, Hammerstad SS, Tomer Y

A preliminary analysis of hepatitis C virus in pancreatic islet cells.

Virol J 2017 Dec 20;14(1):237. Epub 2017 des 20

PMID: 29258547

Lambertini L, Saul SR, Copperman AB, Hammerstad SS, Yi Z, Zhang W, Tomer Y, Kase N

Intrauterine Reprogramming of the Polycystic Ovary Syndrome: Evidence from a Pilot Study of Cord Blood Global Methylation Analysis.

Front Endocrinol (Lausanne) 2017;8():352. Epub 2017 des 18

PMID: 29326659

Hammerstad SS, Stefan M, Blackard J, Owen RP, Lee HJ, Concepcion E, Yi Z, Zhang W, Tomer Y

HEPATITIS C VIRUS E2 PROTEIN INDUCES UPREGULATION OF IL-8 PATHWAYS AND PRODUCTION OF HEAT SHOCK PROTEINS IN HUMAN THYROID CELLS.

J Clin Endocrinol Metab 2016 Nov 18. Epub 2016 nov 18

PMID: 27860532

Tomer Y, Dolan LM, Kahaly G, Divers J, D'Agostino RB, Imperatore G, Dabelea D, Marcovina S, Black MH, Pihoker C, Hasham A, Hammerstad SS, Greenberg DA, Lotay V, Zhang W, Monti MC, Matheis N,

Genome wide identification of new genes and pathways in patients with both autoimmune thyroiditis and type 1 diabetes.

J Autoimmun 2015 Jun;60():32-9. Epub 2015 apr 27

PMID: 25936594

Lee HJ, Li CW, Hammerstad SS, Stefan M, Tomer Y

Immunogenetics of autoimmune thyroid diseases: A comprehensive review.

J Autoimmun 2015 Nov;64():82-90. Epub 2015 jul 30

PMID: 26235382

Hammerstad SS, Grock SF, Lee HJ, Hasham A, Sundaram N, Tomer Y

Diabetes and Hepatitis C: A Two-Way Association.

Front Endocrinol (Lausanne) 2015;6():134. Epub 2015 sep 14

PMID: 26441826

Brown Rosalind S, Lombardi Angela, Hasham Alia, Greenberg David A, Gordon Joshua, Concepcion Erlinda, Hammerstad Sara S, Lotay Vaneet, Zhang Weijia, Tomer Yaron

Genetic analysis in young-age-of-onset Graves' disease reveals new susceptibility loci.

J Clin Endocrinol Metab 2014 Jul;99(7):E1387-91. Epub 2014 mar 31

PMID: 24684463

Hammerstad Sara Salehi, Jahnsen Frode Lars, Tauriainen Sisko, Hyöty Heikki, Paulsen Trond, Norheim Ingrid, Dahl-Jørgensen Knut

Immunological changes and increased expression of myxovirus resistance protein a in thyroid tissue of patients with recent onset and untreated Graves' disease.

Thyroid 2014 Mar;24(3):537-44. Epub 2014 jan 15

PMID: 24032645

Hammerstad Sara Salehi

Hepatitt C og Diabetes: Et Tosidig Forhold

BestPractice Infeksjonsmedisin 2016

Hammerstad SS and Tomer Y

Epidemiology and Genetic factors in Graves' Disease. In book: Graves' Disease A comprehensive Guide for Clinicians

Chapter: 3, Publisher: Springer, Editors: Rebecca S. Bahn, pp.21-37ease and Graves' Ophthalmopathy, April 2015

Hammerstad SS, Ronald Villanueva, Tomer Y

Infection and Autoimmune thyroid Diseases

Book Infection and Autoimmunity Second Edition, Publisher Elsevier Editors Shoenfeld, Agmon-Levin and Rose. February 2015

Deltagere
  • Noel Morgan Prosjektdeltaker
  • Sarah Richardson Prosjektdeltaker
  • Yaron Tomer Prosjektdeltaker
  • Knut Dahl-Jørgensen Hovedveileder
  • Sara Salehi Hammerstad Postdoktorstipendiat (finansiert av denne bevilgning)
  • Frode Lars Jahnsen Forsker
  • Heikki Hyöty Forsker

eRapport er utarbeidet av Sølvi Lerfald og Reidar Thorstensen, Regionalt kompetansesenter for klinisk forskning, Helse Vest RHF, og videreutvikles av de fire RHF-ene i fellesskap, med støtte fra Helse Vest IKT

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