Gene expression profiles in granuloma tissue reveal novel diagnostic markers in sarcoidosis.
Sarcoidosis is an immune-mediated multisystem disease characterized by the formation of non-caseating granulomas. The pathogenesis of sarcoidosis is unclear, with proposed infectious or environmental antigens triggering an aberrant immune response in susceptible hosts. Multiple pro-inflammatory signaling pathways have been implicated in mediating macrophage activation and granuloma formation in sarcoidosis, including IFN-γ/STAT-1, IL-6/STAT-3, and NF-κB. It is difficult to distinguish sarcoidosis from other granulomatous diseases or assess disease severity and treatment response with histopathology alone. Therefore, development of improved diagnostic tools is imperative. Herein, we describe an efficient and reliable technique to classify granulomatous disease through selected gene expression and identify novel genes and cytokine pathways contributing to the pathogenesis of sarcoidosis. We quantified the expression of twenty selected mRNAs extracted from formalin-fixed paraffin embedded (FFPE) tissue (n = 38) of normal lung, suture granulomas, sarcoid granulomas, and fungal granulomas. Utilizing quantitative real-time RT-PCR we analyzed the expression of several genes, including IL-6, COX-2, MCP-1, IFN-γ, T-bet, IRF-1, Nox2, IL-33, and eotaxin-1 and revealed differential regulation between suture, sarcoidosis, and fungal granulomas. This is the first study demonstrating that quantification of target gene expression in FFPE tissue biopsies is a potentially effective diagnostic and research tool in sarcoidosis.
Department of Gastroenterology, Washington University School of Medicine, St. Louis, MO, United States. Electronic address: gchristo@dom.wustl.edu.
George P
GP
Christophi
Department of Pathology, SUNY Upstate Medical University, Syracuse, NY, United States.
Tiffany
T
Caza
Department of Pathology, SUNY Upstate Medical University, Syracuse, NY, United States.
Christopher
C
Curtiss
Department of Internal Medicine, Cleveland Clinic, Cleveland, OH, United States.
Divya
D
Gumber
Department of Microbiology & Immunology, SUNY Upstate Medical University, Syracuse, NY, United States.
Paul T
PT
Massa
Department of Pathology, SUNY Upstate Medical University, Syracuse, NY, United States.
Steve K
SK
Landas
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D054413:Chemokine CCL11 / Q000235:genetics / Q000378:metabolism; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D002648:Child; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D005819:Genetic Markers; D006099:Granuloma / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism; D000067596:Interleukin-33; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007378:Interleukins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D012507:Sarcoidosis / Q000175:diagnosis* / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D013048:Specimen Handling; D059467:Transcriptome; D015854:Up-Regulation; D055815:Young Adult
C515456:CCL11 protein, human; C111835:CCL2 protein, human; D054413:Chemokine CCL11; D018932:Chemokine CCL2; D005819:Genetic Markers; C506536:IL33 protein, human; D000067596:Interleukin-33; D015850:Interleukin-6; D007378:Interleukins; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D007371:Interferon-gamma; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases
Cytokines; Formalin-fixed paraffin embedded tissue; Gene expression; Granuloma; Pathology autoimmunity; RT-PCR; Reactive oxygen species; Sarcoidosis; Transcription factor