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JAPANESE SUMMARY (和文要旨)

The Association of Thyroglobulin Single Nucleotide Polymorphism with Miniature DachshundsSpecific Inflammatory Colorectal Polyps and its Involvement in Interleukin-6 Amplifier Induced

Chronic Inflammation

(ミニチュアダックスフンドに特異的な炎症性結直腸ポリープに関連するサイログロブ

リンの一塩基多型とインターロイキン 6 増幅回路に誘導される慢性炎症への関与)

犬の炎症性結直腸ポリープ（Inflammatory colorectal polyp, ICRP）は、本邦

のミニチュアダックスフンド（Miniature Dachshund, MD）に好発する特徴的な炎症性

腸疾患である。症例は主に血便、しぶりおよび粘液便などの臨床徴候を呈し、下部消化

管内視鏡検査では下行結腸から直腸の粘膜において、出血を伴う孤立性または多発性の

炎症性ポリープが観察される。病理組織検査では好中球、マクロファージ、リンパ球浸

潤を伴う杯細胞の過形成を認め、この組織像および免疫抑制療法に良好な反応を示すこ

とから、ICRP の病態には遺伝的な免疫異常が関与していることが示唆されている。近

年、病態解明を目的とした ICRP の病変部における炎症性サイトカインの発現を調べた

研究が多数報告されているものの、MD に特異的かつ炎症を誘導する遺伝子は明らかに

なっていない。そこで本研究は、MD に特異的かつ炎症を引き起こす遺伝子を明らかに

し、その炎症機序を解明することを目的とした。

第１章では、先行研究で特定されたリスク遺伝子データを再検討した。MD に特

異的な ICRP の疾患に関連するより多くの感受性遺伝子を検出するため、非 MD、MDICRP および MD-Control 群それぞれにサンプルを追加した。以前の研究と比較して、TG

(exon22:c.C4567T:p.R1523W)と FBN1 (exon10:c.C1205T:p.P402L)の SNP が追加で検出

され、TG に ICRP の発症機序との有意な関連性が示された。

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第２章では、マウスの疾患モデルや慢性炎症、ならびに医学領域の自己免疫疾患

で確立されている炎症経路であるインターロイキン 6 増幅回路(IL-6 アンプ)と、第１

章で同定された ICRP の病因候補遺伝子の関与を評価した。サイレンシング RNA を用い

て TG をノックダウンしたところ、非免疫細胞系である H4 細胞において IL-6 mRNA の発

現が抑制されることが証明された。さらに、非免疫細胞に対して recombinant TG を処

理したところ、濃度依存的な IL-6 mRNA の発現が顕著に誘導された。また、TG は IL-6

アンプを介する NF-κB の標的分子であったため、TG のポジティブフィードバックルー

プが炎症を増悪する可能性が示唆された。これらの結果から、TG は、従来、甲状腺ホ

ルモン生成にのみ関与するタンパク質と考えられていたものの、免疫細胞以外の慢性炎

症に影響を与える IL-6 アンプの主要な調節遺伝子であることが示された。

最後に、第３章では ICRP に罹患した MD において全身的あるいは局所的な TG の

発現を調べるため、末梢血の TG 濃度および非炎症性大腸粘膜における TG の発現を

ELISA キットと qPCR で解析した。サンプルは ICRP 罹患犬とコントロール犬の血清およ

び結腸組織とした。ポリープ状病変には、免疫細胞の浸潤による過剰な炎症が引き起こ

されていると予想されたため、内視鏡検査と病理組織学的検査により正常と診断された

大腸粘膜サンプルを qPCR の発現解析の対象とした。末梢血の TG 濃度には、ICRP 罹患

群およびコントロール群で有意な差は認められなかった。さらに成犬および子犬のアレ

ル群間での比較においても明らかな差は認められなかった。一方で結腸組織において

は、Taqman プローブ定量 PCR を用いた非炎症性大腸粘膜中の TG mRNA 発現量には、T/T

および C/C アレル群間で有意な差が見られた。さらに、IL-6 アンプも ICRP 罹患群の

T/T リスクアレルを有する場合に活性化されることが証明され、NF-κB の標的である

IL-6 および CCL2 の発現が増加することが示された。以上のことから、TG 発現が IL-6

アンプの活性化を誘導する上で重要であり、ICRP の発症機序に関与していることが示

唆された。

本研究では、TG c.4567C>T SNP の機能評価を通じて、MD の犬種特異性と ICRP の

遺伝的関連性が ICRP の炎症性病態の確立につながることが証明された。さらに興味深

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いことに、非典型的な炎症分子である TG が慢性炎症に関連していることが示された。

これは医学領域で甲状腺疾患を有する患者に IBD が高頻度で発生することの根拠となる

可能性がある。また、本研究を通じて、動物の疾患の研究は、汎動物学の考え方に沿っ

て、人の疾患の研究に発展する可能性があることも示唆された。

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