Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing
High-throughput RNA sequencing (RNA-seq) uses massive parallel sequencing technology, allowing the unbiased analysis of genome-wide transcription levels and tumor mutation status. Immunoglobulin G4-related ophthalmic disease (IgG4-ROD) is a fibroinflammatory disease characterized by the enlargement...
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doaj-8d2851e9c5c541ef92768a31d74a1a082020-11-25T03:52:36ZengMDPI AGJournal of Clinical Medicine2077-03832020-10-0193458345810.3390/jcm9113458Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA SequencingMasaki Asakage0Yoshihiko Usui1Naoya Nezu2Hiroyuki Shimizu3Kinya Tsubota4Kazuhiko Umazume5Naoyuki Yamakawa6Tomohiro Umezu7Hirotsugu Suwanai8Masahiko Kuroda9Hiroshi Goto10Department of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Molecular Pathology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Diabetes, Metabolism and Endocrinology, Tokyo Medical University, 6-7-1 Nishi-shinjuku Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Molecular Pathology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanDepartment of Ophthalmology, Tokyo Medical University, 6-7-1 Nishi-shinjuku, Shinjuku-ku, Tokyo 160-0023, JapanHigh-throughput RNA sequencing (RNA-seq) uses massive parallel sequencing technology, allowing the unbiased analysis of genome-wide transcription levels and tumor mutation status. Immunoglobulin G4-related ophthalmic disease (IgG4-ROD) is a fibroinflammatory disease characterized by the enlargement of the ocular adnexal tissues. We analyzed RNA expression levels via RNA-seq in the biopsy specimens of three patients diagnosed with IgG4-ROD. Mucosa-associated lymphoid tissue (MALT) lymphoma, reactive lymphoid hyperplasia (RLH), normal lacrimal gland tissue, and adjacent adipose tissue were used as the controls (<i>n</i> = 3 each). RNA-seq was performed using the NextSeq 500 system, and genes with |fold change| ≥ 2 and <i>p</i> < 0.05 relative to the controls were defined as differentially expressed genes (DEGs) in IgG4-ROD. To validate the results of RNA-seq, real-time polymerase chain reaction (PCR) was performed in 30 IgG4-ROD and 30 orbital MALT lymphoma tissue samples. RNA-seq identified 35 up-regulated genes, including matrix metallopeptidase 12 (MMP12) and secreted phosphoprotein 1 (SPP1), in IgG4-ROD tissues when compared to all the controls. Many pathways related to the immune system were included when compared to all the controls. Expressions of MMP12 and SPP1 in IgG4-ROD tissues were confirmed by real-time PCR and immunohistochemistry. In conclusion, we identified novel DEGs, including those associated with extracellular matrix degradation, fibrosis, and inflammation, in IgG4-ROD biopsy specimens. These data provide new insights into molecular pathogenetic mechanisms and may contribute to the development of new biomarkers for diagnosis and molecular targeted drugs.https://www.mdpi.com/2077-0383/9/11/3458IgG4-related diseaseIgG4-related ophthalmic diseaseRNA sequencingMMP12SPP1orbital lymphoproliferative disorders |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Masaki Asakage Yoshihiko Usui Naoya Nezu Hiroyuki Shimizu Kinya Tsubota Kazuhiko Umazume Naoyuki Yamakawa Tomohiro Umezu Hirotsugu Suwanai Masahiko Kuroda Hiroshi Goto |
spellingShingle |
Masaki Asakage Yoshihiko Usui Naoya Nezu Hiroyuki Shimizu Kinya Tsubota Kazuhiko Umazume Naoyuki Yamakawa Tomohiro Umezu Hirotsugu Suwanai Masahiko Kuroda Hiroshi Goto Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing Journal of Clinical Medicine IgG4-related disease IgG4-related ophthalmic disease RNA sequencing MMP12 SPP1 orbital lymphoproliferative disorders |
author_facet |
Masaki Asakage Yoshihiko Usui Naoya Nezu Hiroyuki Shimizu Kinya Tsubota Kazuhiko Umazume Naoyuki Yamakawa Tomohiro Umezu Hirotsugu Suwanai Masahiko Kuroda Hiroshi Goto |
author_sort |
Masaki Asakage |
title |
Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing |
title_short |
Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing |
title_full |
Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing |
title_fullStr |
Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing |
title_full_unstemmed |
Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing |
title_sort |
comprehensive gene analysis of igg4-related ophthalmic disease using rna sequencing |
publisher |
MDPI AG |
series |
Journal of Clinical Medicine |
issn |
2077-0383 |
publishDate |
2020-10-01 |
description |
High-throughput RNA sequencing (RNA-seq) uses massive parallel sequencing technology, allowing the unbiased analysis of genome-wide transcription levels and tumor mutation status. Immunoglobulin G4-related ophthalmic disease (IgG4-ROD) is a fibroinflammatory disease characterized by the enlargement of the ocular adnexal tissues. We analyzed RNA expression levels via RNA-seq in the biopsy specimens of three patients diagnosed with IgG4-ROD. Mucosa-associated lymphoid tissue (MALT) lymphoma, reactive lymphoid hyperplasia (RLH), normal lacrimal gland tissue, and adjacent adipose tissue were used as the controls (<i>n</i> = 3 each). RNA-seq was performed using the NextSeq 500 system, and genes with |fold change| ≥ 2 and <i>p</i> < 0.05 relative to the controls were defined as differentially expressed genes (DEGs) in IgG4-ROD. To validate the results of RNA-seq, real-time polymerase chain reaction (PCR) was performed in 30 IgG4-ROD and 30 orbital MALT lymphoma tissue samples. RNA-seq identified 35 up-regulated genes, including matrix metallopeptidase 12 (MMP12) and secreted phosphoprotein 1 (SPP1), in IgG4-ROD tissues when compared to all the controls. Many pathways related to the immune system were included when compared to all the controls. Expressions of MMP12 and SPP1 in IgG4-ROD tissues were confirmed by real-time PCR and immunohistochemistry. In conclusion, we identified novel DEGs, including those associated with extracellular matrix degradation, fibrosis, and inflammation, in IgG4-ROD biopsy specimens. These data provide new insights into molecular pathogenetic mechanisms and may contribute to the development of new biomarkers for diagnosis and molecular targeted drugs. |
topic |
IgG4-related disease IgG4-related ophthalmic disease RNA sequencing MMP12 SPP1 orbital lymphoproliferative disorders |
url |
https://www.mdpi.com/2077-0383/9/11/3458 |
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