胃癌と十二指腸潰瘍-ピロリ菌感染の場を起点として解く異なる二つの胃疾患発症機序

• Project/Area Number: 21K07898
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Oita University
• Principal Investigator: 赤田 純子 大分大学, 医学部, 助教 (30346548)
• Co-Investigator(Kenkyū-buntansha): 山岡 吉生 大分大学, 医学部, 教授 (00544248) ; 松本 昂 大分大学, 医学部, 助教 (50609667) ; 塚本 善之 大分大学, 医学部, 助教 (00433053) ; 村上 和成 大分大学, 医学部, 教授 (00239485)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2023: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2022: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2021: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: ヘリコバクター・ピロリ / 胃癌 / 十二指腸潰瘍 / GWAS解析 / 胃オルガノイド

Outline of Research at the Start

東アジアの胃癌死亡率は世界平均の2倍近くと、胃癌死の低減へは道半ばである。胃癌の主要な誘導因子はHelicobacter pylori（ピロリ菌）である。我々は東アジアピロリ菌株を用い、対照的な胃疾患である胃癌と十二指腸潰瘍にそれぞれ由来する菌株を、ゲノムワイド遺伝子関連解析にて比較し、胃癌株に優位に関連する１遺伝子変異（SNP）を持つ11遺伝子を選択した。本研究では、ピロリ菌感染の場であるヒト胃粘膜組織、および胃オルガノイド感染実験系を用いて、十二指腸潰瘍および胃癌発症に関与するピロリ菌病原体因子と、関連する宿主因子を明らかにする。

Outline of Annual Research Achievements

胃癌の主要な誘導因子はHelicobacter pylori（ピロリ菌）である。我々はこれまでに、東アジアピロリ菌株を用い、対照的な胃疾患である胃癌（GC）と十二指腸潰瘍（DU）に由来する菌株をゲノムワイド遺伝子関連解析（GWAS）にて比較し、胃癌株に優位に関連する１遺伝子変異（SNP）を持つ11遺伝子を選択した。新たに導入した胃オルガノイド感染実験系を用いて、十二指腸潰瘍および胃癌発症に関与するピロリ菌病原体因子と、関連する宿主因子を明らかにしてゆく。

今年度は、昨年度確立した日本人胃幽門部由来の二次元胃オルガノイドを用いた感染実験を行い、感染オルガノイドのmRNAを回収しアレープレートを用いたRT-qPCR発現解析を行った。その結果、ピロリ菌感染により、サイトカイン遺伝子５種、DNA修復機構遺伝子４種、がん関連遺伝子10種に２倍以上の発現上昇を認めた。候補遺伝子については確認実験を行うと共に、感染実験のタイムコースなど詳細な実験条件をなお設定中である。そのために胃上皮系AGS細胞感染実験も並行して進めたが、mRNA実験条件最適化によりサイトカイン遺伝子11種においては感染細胞では10倍以上の発現上昇を認めた。さらに解析を進めてゆく。

また、ピロリ菌病原因子解析においては、GWAS-SNPを持つ11遺伝子のうちのdsbG遺伝子について26695株を親株として用い、dsbG遺伝子破壊株、GC-SNP含有dsbG遺伝子相補株、DU-SNP含有dsbG遺伝子相補株を作成し、酸性条件下でのみ遊走能に差があることを明らかにした。In vitro機能評価実験と共に、病原性の評価がしやすいIn vivoスナネズミ感染実験に移行することも視野に入れ、スナネズミ馴化東アジア株TN２株を用いて、dsbG遺伝子改変株を作成中である。またdsbG以外にも３つのGWAS抽出遺伝子改変株を作成中である。

Research Products

• [Int'l Joint Research] Cho Ray Hospital/108 Military Central Hospital(ベトナム)
• [Int'l Joint Research] Mongolian National Univ. of Med. Sci.(モンゴル)
• [Int'l Joint Research] Universitias Airlangga(インドネシア)
• [Int'l Joint Research] Universty of Mbujimai(コンゴ民主共和国)
• [Int'l Joint Research] Cho Ray Hospital(ベトナム)
• [Int'l Joint Research] Mongolian National Univ. of Med. Sci.(モンゴル)
• [Int'l Joint Research] Universitas Airlangga(インドネシア)
• [Int'l Joint Research] University of Mbujima(コンゴ民主共和国)
• [Int'l Joint Research] Cho Ray Hospital/108 Military Central Hospital(ベトナム)
• [Journal Article] Comparative genomics of two Vietnamese Helicobacter pylori strains, CHC155 from a non-cardia gastric cancer patient and VN1291 from a duodenal ulcer patient. (2023)
  • Author(s): Phuc BH, Tuan VP, Binh TT, Tung PH, Tri TD, Dung HDQ, Thuan NPM, Fauzia KA, Tshibangu-Kabamba E, Alfaray RI, Saruuljavkhlan B, Matsumoto T, Akada J, Yamaoka Y.
  • Journal Title: Sci Rep Volume: 13 Issue: 1 Pages: 8869-8869
  • DOI: 10.1038/s41598-023-35527-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Study of Helicobacter pylori Isolated from a High-Gastric-Cancer-Risk Population: Unveiling the Comprehensive Analysis of Virulence-Associated Genes including Secretion Systems, and Genome-Wide Association Study (2023)
  • Author(s): Saruuljavkhlan B, Alfaray RI, Oyuntsetseg K, Gantuya B, Khangai A, Renchinsengee N, Matsumoto T, Akada J, Azzaya D, Davaadorj D, Yamaoka Y.
  • Journal Title: Cancers (Basel) Volume: 15 Issue: 18 Pages: 4528-4528
  • DOI: 10.3390/cancers15184528
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Helicobacter pyloriの環境に適応する病原因子とはなんですか? (2023)
  • Author(s): 赤田 純子
  • Journal Title: Helicobacter Reserch Volume: 27 Pages: 129-135
• [Journal Article] The Helicobacter pylori Genome Project: insights into H. pylori population structure from analysis of a worldwide collection of complete genomes (2023)
  • Author(s): Kaisa Thorell, Zilia Y. Munoz-Ramirez, Difei Wang, Santiago Sandoval-Motta, Rajiv Boscolo Agostini, Silvia Ghirotto, Roberto C. Torres, HpGP Research Network, Daniel Falush, M. Constanza Camargo & Charles S. Rabkin
  • Journal Title: Nature Communications Volume: 14 Issue: 1 Pages: 8184-8184
  • DOI: 10.1038/s41467-023-43562-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Utilization of an Automated Latex Agglutination Turbidity Assay for Assessing Gastric Mucosal Alteration during Helicobacter pylori Infection (2023)
  • Author(s): Khangai Ayush、Akada Junko、Saruuljavkhlan Batsaikhan、Gantuya Boldbaatar、Azzaya Dashdorj、Oyuntsetseg Khasag、Davaadorj Duger、Uchida Tomohisa、Matsumoto Takashi、Yamaoka Yoshio
  • Journal Title: Gut and Liver Volume: 18 Issue: 1 Pages: 60-69
  • DOI: 10.5009/gnl220464
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 日本国内より分離されたHelicobacter pylori の集団構造解析 (2023)
  • Author(s): 友成航平, Batsaikhan S, Alfaray, RI, Fauzia KA, 松成修, 赤田純子, 松本昂, 珍田大輔, 下山克, 金城渚, 金城徹, 金城福則, 山岡吉生
  • Journal Title: 大分県医学会雑誌 Volume: 29 Pages: 100-108
  • Int'l Joint Research
• [Journal Article] Helicobacter pylori genomes reveal Paleolithic human migration to the east end of Asia (2022)
  • Author(s): Suzuki Rumiko、Saitou Naruya、Matsuari Osamu、Shiota Seiji、Matsumoto Takashi、Akada Junko、Kinjo Nagisa、Kinjo Fukunori、Teruya Kuniko、Shimoji Makiko、Shiroma Akino、Kato Mototsugu、Satou Kazuhito、Hirano Takashi、Asaka Masahiro、Kryukov Kirill、Moodley Yoshan、Yamaoka Yoshio
  • Journal Title: iScience Volume: 25 Issue: 7 Pages: 104477-104477
  • DOI: 10.1016/j.isci.2022.104477
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genome-wide association study of gastric cancer- and duodenal ulcer-derived Helicobacter pylori strains reveals discriminatory genetic variations and novel oncoprotein candidates (2021)
  • Author(s): Tuan V P、Yahara K、Dung H D Q、Binh T T、Huu T P、Tri T D、Thuan N P M、Khien V V、Trang T T H、Phuc B H、Tshibangu-Kabamba E、Matsumoto T、Akada J、Suzuki R、Okimoto T、Kodama M、Murakami K、Yano H、Fukuyo M、Takahashi N、Kato M、Nishiumi S、Azuma T、Ogura Y、Hayashi T、Toyoda A、Kobayashi I、Yamaoka Y
  • Journal Title: Microbial Genomics Volume: 7 Issue: 11 Pages: 1-17
  • DOI: 10.1099/mgen.0.000680
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Serum Helicobacter pylori antibody reactivity in seven Asian countries using an automated latex aggregation turbidity assay (2021)
  • Author(s): Akada J,Tshibangu‐K,Tuan V,Kurogi S,Matsuo Y,Ansari S,Doohan D,Phuc B,Subsomwong P,Waskito L,Binh T,Nguyen L,Khien V,Dung H,Miftahussurur M,Syam A,Tshering L,Vilaichone R,Mahachai V,Ratanachu‐ek T,Shrestha P,Yee T,Htet K,Aftab H,Matsuhisa T,Uchida T,Okimoto T,Mizukami K,Kodama M,Murakami K,Takahashi N,Yamaoka Yoshio
  • Journal Title: Journal of Gastroenterology and Hepatology Volume: - Issue: 8 Pages: 1-10
  • DOI: 10.1111/jgh.15467
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Helicobacter pylori type 4 secretion systems as gastroduodenal disease markers (2021)
  • Author(s): Phuc Bui Hoang、Tuan Vo Phuoc、Dung Ho Dang Quy、Binh Tran Thanh、Tung Pham Huu、Tri Tran Dinh、Thuan Ngo Phuong Minh、Van Khien Vu、Trang Tran Thi Huyen、Akada Junko、Matsumoto Takeshi、Yamaoka Yoshio
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 4584-4584
  • DOI: 10.1038/s41598-021-83862-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Entroccus Hirae Attenuates chemokine and Induces Dna Damage In Gastric Epithelial Cell (2023)
  • Author(s): Saruuljavkhlan B, Gantuya B, Alfaray RI, Akada J, Matsumoto T, Tsukamoto Y, Oyuntsetseg K, Yamaoka Y
  • Organizer: DDW USA
  • Int'l Joint Research
• [Presentation] Investigation of Low－Grade Intestinal Metaplasia From Clinical Dyspepsia Patients INA Country With Low-Incidence of Gastric Cancer:From Human to Helicobactor pylori Perspective (2023)
  • Author(s): Alfaray RI, Miftahussurur M, Saruuljavkhlan B, Fauzia KA, Utomo, DH, Akada J, Matsumoto T, Yamaoka Y
  • Organizer: DDW USA
  • Int'l Joint Research
• [Presentation] Amoxicillin resistance in Helicobacter pylori: Identification of new resistance-conferring mutations in the penicillin-binding protein 1A (2023)
  • Author(s): Cimuanga-Mukanya A, Tshibangu-Kabamba E, Ngoma-Kisoko PdeJ , Disashi-Tumba G, Matsumoto T,Akada J and Yamaoka Y
  • Organizer: The 19th Korea-Japan H. pylori Joint Symposium.
  • Int'l Joint Research
• [Presentation] Utilization of an Automated Latex Agglutination Turbidity Assay for Assessing Gastric Mucosal Alteration during Helicobacter pylori Infection (2023)
  • Author(s): Khangai A, Akada J, Azzaya D, Batsaikhan S, Gantuya B, Oyuntsetseg K, Davaadaorj D, Matsumoto T, Uchida T, Akada J, and Yamaoka Y.
  • Organizer: The 19th Korea-Japan H. pylori Joint Symposium.
  • Int'l Joint Research
• [Presentation] The impact of mobile genetic elemants and toxin-antitoxin system on Helicobacter pylori adaptability and antimicrobial resistance: A wisdom from Japan-South Korea whole-genome sequence collection. (2022)
  • Author(s): Alfaray RI, Batsaikhan S, Fauzia KA, Akada J, Matsumoto T, Yamaoka Y.
  • Organizer: The 18th Japan-Korea H. pylori Joint Symposium
  • Int'l Joint Research
• [Presentation] Identification of the Helicobacter pylori genomic islands in a high gastric cancer region. (2022)
  • Author(s): Batsaikhan S, Alfaray RI, Matsumoto T, Akada J, Yamaoka Y.
  • Organizer: The 18th Japan-Korea H. pylori Joint Symposium
  • Int'l Joint Research
• [Presentation] 胃癌特異的一塩基多型を有するピロリ菌dsbG遺伝子の比較解析 (2021)
  • Author(s): 赤田 純子
  • Organizer: 第7回日本ヘリコバクター学会学術集会