Innovative therapy for lung cancer: prophylaxis and therapy by intratracheal delivery of probiotics harboring immune checkpoint-blocking single-chain variable fragment

• Project/Area Number: 19KK0208
• Research Category: Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 50:Oncology and related fields
• Research Institution: Shinshu University
• Principal Investigator: 佐藤 隆 信州大学, 先鋭領域融合研究群バイオメディカル研究所, 特任教授 (70510436)
• Co-Investigator(Kenkyū-buntansha): 下里 剛士 信州大学, 学術研究院農学系, 教授 (00467200) ; 荻田 佑 信州大学, 学術研究院農学系, 助教 (50738010) ; 重盛 駿 信州大学, 先鋭領域融合研究群バイオメディカル研究所, 助教(特定雇用) (90803487) ; 生井 楓 東北大学, 農学研究科, 特任助教 (60893857)
• Project Period (FY): 2019-10-07 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2022: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2021: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
• Keywords: 遺伝子組換え乳酸菌 / 肺がん / 経気道治療 / 免疫チェックポイント阻害 / 生物資源医薬 / 経気道的標的治療 / 肺癌

Outline of Research at the Start

本研究は、気道粘膜免疫を標的とする「微生物由来合成核酸の経気道治療の研究」と、「有用微生物である乳酸菌由来高分子・抗体医薬の経気道治療の研究」成果を基盤とし、免疫チェックポイント阻害作用を有する乳酸菌による肺癌発癌・進展予防効果を検証する内容である。本研究は、早期の臨床試験への導出を目的として、癌研究と治療開発で世界最先端の米国テキサス大学MDアンダーソンがんセンターとの国際共同研究として計画した。申請者が所属する信州大学の乳酸菌研究のエキスパートと若手研究者の参画に加えて、国際的に高い信頼と臨床導出の実績がある癌専門研究機関の参画により、早期に次世代治療として確立を目指す内容である。

Outline of Annual Research Achievements

本研究は「生理活性標的物質を分泌する乳酸菌」の「経気道投与」による呼吸器疾患治療の可能性を提唱する研究成果（Antioxidants 2020;9:1049）を基盤とし、「免疫チェックポイント阻害乳酸菌」による肺癌発癌・進展予防効果を検証する国際共同研究である。信州大学での同所性移植肺がんモデルや転移性肺腫瘍モデルでの生存延長効果を確認し、米国テキサス大学・MDアンダーソンがんセンターでの自然発癌モデルの発癌予防効果の検証を踏まえて、臨床導出を目指す内容である。これまでに、免疫チェックポイント阻害乳酸菌としてLactococcus lactis NZ-9000株への標的遺伝子発現ベクター導入により、1）Programmed cell death (PD)- 1 阻害作用を有する低分子一本鎖可変抗体フラグメント（single chain variable fragment：scFv）、2）PD-L1 scFv、さらに3）
Cytotoxic T-Lymphocyte-associated protein (CTLA)-4 scFv産生組換え乳酸菌を構築し、生理的活性を確認した。これらの組換え乳酸菌の動物モデルを用いた生存率の検証で、PD-1scFv産生組換え乳酸菌は同所性移植肺がんモデルの有意な生存延長効果を示した（PD-1scFv投与群 vs ベクターコントロール（VC）投与群：67% vs 0%）。また、PD-L1scFv産生組換え乳酸菌においても転移性肺腫瘍モデルで有意な生存延長効果を確認した（PD-L1scFv投与群 vs VC投与群：62.5% vs 0%）。最終年度には、国際共同研究ワークショップを開催し、組換え乳酸菌による肺癌治療ストラテジーの構築への取り組みを紹介するとともに、海外日本人研究者との新たな連携を構築し、国際共同研究を加速する機会の創出に取り組んだ。

Research Products

• [Int'l Joint Research] テキサス大学(米国)
• [Int'l Joint Research] MD Anderson Cancer Center(米国)
• [Int'l Joint Research] テキサス大学(米国)
• [Int'l Joint Research] テキサス大学MDアンダーソンがんセンター(米国)
• [Int'l Joint Research] テキサス大学MDアンダーソンがんセンター(米国)
• [Journal Article] 組換え乳酸菌による肺がん免疫治療 (2024)
  • Author(s): 佐藤 隆
  • Journal Title: Precision Medicine Volume: 7 Pages: 554-558
• [Journal Article] 組換え乳酸菌による医薬・ワクチン開発 (2024)
  • Author(s): 佐藤 隆
  • Journal Title: BIO Clinica Volume: 39 Pages: 531-535
• [Journal Article] Development of a Single-Chain Fragment Variable that Binds to the SARS-CoV-2 Spike Protein Produced by Genetically Modified Lactic Acid Bacteria (2024)
  • Author(s): Oshima Suzuka、Namai Fu、Sato Takashi、Shimosato Takeshi
  • Journal Title: Molecular Biotechnology Volume: 66 Issue: 1 Pages: 151-160
  • DOI: 10.1007/s12033-023-00741-y
  • Peer Reviewed / Open Access
• [Journal Article] 組換え乳酸菌による肺癌免疫治療 (2023)
  • Author(s): 佐藤 隆
  • Journal Title: BIO Clinica Volume: 38 Pages: 638-642
• [Journal Article] Oral administration of <scp><i>Brevibacterium linens</i></scp> from washed cheese increases the proportions of short‐chain fatty acid‐producing bacteria and lactobacilli in the gut microbiota of mice (2023)
  • Author(s): Murakami Aito、Toyomoto Koharu、Namai Fu、Sato Takashi、Fujii Tadashi、Tochio Takumi、Shimosato Takeshi
  • Journal Title: Animal Science Journal Volume: 94 Issue: 1
  • DOI: 10.1111/asj.13905
  • Peer Reviewed / Open Access
• [Journal Article] Development of fluorescence-labeled antibody for immune checkpoint inhibitor using engineered probiotics (2023)
  • Author(s): Namai Fu、Sumiya Shunsuke、Nomura Natsumi、Sato Takashi、Shimosato Takeshi
  • Journal Title: AMB Express Volume: 13 Issue: 1 Pages: 4-4
  • DOI: 10.1186/s13568-023-01509-y
  • Peer Reviewed / Open Access
• [Journal Article] Free feeding of CpG-oligodeoxynucleotide particles prophylactically attenuates allergic airway inflammation and hyperresponsiveness in mice (2021)
  • Author(s): Okajima, T., Shigemori, S., Namai, F., Ogita, T., Sato, T. & Shimosato, T.
  • Journal Title: Frontiers in Immunology Volume: 12 Pages: 738041-738041
  • DOI: 10.3389/fimmu.2021.738041
  • Peer Reviewed / Open Access
• [Journal Article] Oral administration of Flavonifractor plautii, a bacteria increased with green tea consumption, promotes recovery from acute colitis in mice via suppression of IL-17 (2021)
  • Author(s): Ayane Mikami, Tasuku Ogita, Fu Namai, Suguru Shigemori, Takashi Sato, Takeshi Shimosato
  • Journal Title: Frontiers in Nutrition Volume: 7 Pages: 610946-610946
  • DOI: 10.3389/fnut.2020.610946
  • Peer Reviewed / Open Access
• [Journal Article] Construction of Genetically Modified Lactococcus lactis Producing Anti-human-CTLA-4 Single-Chain Fragment Variable (2020)
  • Author(s): Namai Fu、Murakami Aito、Ueda Asami、Tsukagoshi Masami、Shigemori Suguru、Ogita Tasuku、Sato Takashi、Shimosato Takeshi
  • Journal Title: Molecular Biotechnology Volume: 62 Issue: 11-12 Pages: 572-579
  • DOI: 10.1007/s12033-020-00274-8
  • Peer Reviewed
• [Journal Article] Nasally Administered Lactococcus lactis Secreting Heme Oxygenase-1 Attenuates Murine Emphysema (2020)
  • Author(s): Yumoto Kentaro、Sato Takashi、Nakashima Kentaro、Namai Fu、Shigemori Suguru、Shimosato Takeshi、Kaneko Takeshi
  • Journal Title: Antioxidants Volume: 9 Issue: 11 Pages: 1049-1049
  • DOI: 10.3390/antiox9111049
  • Peer Reviewed / Open Access
• [Journal Article] Construction of genetically modified Lactococcus lactis that produces bioactive anti-interleukin-4 single-chain fragment variable (2020)
  • Author(s): Namai Fu、Shigemori Suguru、Ogita Tasuku、Sato Takashi、Shimosato Takeshi
  • Journal Title: Molecular Biology Reports Volume: 47 Issue: 9 Pages: 7039-7047
  • DOI: 10.1007/s11033-020-05765-0
  • Peer Reviewed
• [Journal Article] Microbial therapeutics for acute colitis based on genetically modified Lactococcus lactis hypersecreting IL-1Ra in mice (2020)
  • Author(s): Namai Fu、Shigemori Suguru、Ogita Tasuku、Sato Takashi、Shimosato Takeshi
  • Journal Title: Experimental & Molecular Medicine Volume: 52 Issue: 9 Pages: 1627-1636
  • DOI: 10.1038/s12276-020-00507-5
  • Peer Reviewed / Open Access
• [Journal Article] Ribosome-Engineered Lacticaseibacillus rhamnosus Strain GG Exhibits Cell Surface Glyceraldehyde-3-Phosphate Dehydrogenase Accumulation and Enhanced Adhesion to Human Colonic Mucin (2020)
  • Author(s): Ishida Minori、Namai Fu、Shigemori Suguru、Kajikawa Shoko、Tsukagoshi Masami、Sato Takashi、Ogita Tasuku、Shimosato Takeshi
  • Journal Title: Applied and Environmental Microbiology Volume: 86 Issue: 20
  • DOI: 10.1128/aem.01448-20
  • Peer Reviewed / Open Access
• [Journal Article] Oral administration of Flavonifractor plautii attenuates inflammatory responses in obese adipose tissue (2020)
  • Author(s): Ayane Mikami, Tasuku Ogita, Fu Namai, Suguru Shigemori, Takashi Sato, Takeshi Shimosato
  • Journal Title: Molecular Biology Reports Volume: 47 Issue: 9 Pages: 6717-6725
  • DOI: 10.1007/s11033-020-05727-6
  • Peer Reviewed
• [Journal Article] The clinical significance of CXCL16 in the treatment of advanced non‐small cell lung cancer (2020)
  • Author(s): Shibata Yuji、Kobayashi Nobuaki、Sato Takashi、Nakashima Kentaro、Kaneko Takeshi
  • Journal Title: Thoracic Cancer Volume: 11 Issue: 5 Pages: 1258-1264
  • DOI: 10.1111/1759-7714.13387
  • Peer Reviewed / Open Access
• [Journal Article] Oral Administration of Flavonifractor plautii Strongly Suppresses Th2 Immune Responses in Mice (2020)
  • Author(s): Ogita Tasuku、Yamamoto Yoshinari、Mikami Ayane、Shigemori Suguru、Sato Takashi、Shimosato Takeshi
  • Journal Title: Frontiers in Immunology Volume: 11 Pages: 379-379
  • DOI: 10.3389/fimmu.2020.00379
  • Peer Reviewed / Open Access
• [Presentation] Microbial therapeutics for lung cancer based on immune-checkpoint inhibitory scFv-producing engineered lactic acid bacteria (2023)
  • Author(s): Fu Namai, Haruki Kitazawa, Takashi Sato, Takeshi Shimosato
  • Organizer: FEMS (Federation of European Microbiological Societies) 2023
  • Int'l Joint Research
• [Presentation] Intranasal Administration of Genetically Modified Lactococcus Lactis Producing PD-L1 scFv Improves Survival in Lung Cancer Model Mice (2023)
  • Author(s): Natsumi Nomura, Fu Namai, Takashi Sato, Takeshi Shimosato
  • Organizer: ATS (American Thoracic Society) 2023 International Conference
  • Int'l Joint Research
• [Presentation] Effect of Local Intratracheal Therapy Using Particleized Nintedanib in a Murine Model of Pulmonary Fibrosis (2023)
  • Author(s): Kaho Suzuki, Fu Namai, Natsumi Nomura, Yuichi Sato, Akihiro Kotsugai, Takeshi Shimosato, Takashi Sato
  • Organizer: ATS (American Thoracic Society) 2023 International Conference
  • Int'l Joint Research
• [Presentation] Intranasal Administration of Genetically Modified Lactococcus Lactis Producing PD-L1scFv Improves Survival in Lung Cancer Model Mice (2023)
  • Author(s): Nomura N, Namai F, Sato T, Shimosato T
  • Organizer: American Thoracic Society 2023 International Conference
  • Int'l Joint Research
• [Presentation] Intranasal-administrable microbial therapeutics for lung cancer based on immune- checkpoint inhibitory scFv-producing genetically modified lactic acid bacteria (2023)
  • Author(s): Namai F, Sato T, Shimosato T
  • Organizer: Federation of European Microbiological Societies 2023
  • Int'l Joint Research
• [Presentation] Intratracheal therapy using oligodeoxynucleotides and/or genetically modified lactic acid bacteria-derived biological molecules (2022)
  • Author(s): Takashi Sato
  • Organizer: Korean Society of Toxicology Spring International Symposium
  • Int'l Joint Research / Invited
• [Presentation] 1.抗CTLA-4低分子抗体を産生するLactococcus lactis組換え体の構築 (2019)
  • Author(s): 村上愛斗, 生井楓, 上田麻未，重盛駿, 荻田佑, 佐藤隆, 下里剛士
  • Organizer: 2019酪農科学シンポジウム
• [Presentation] 2.PD-1/PD-L1チェックポイントの阻害に働く乳酸菌組み換え体の構築と肺がんモデルマウスを用いた検証 (2019)
  • Author(s): 上田麻未，生井楓，村上愛斗，重盛駿，荻田佑，佐藤隆，下里剛士
  • Organizer: 2019酪農科学シンポジウム
• [Presentation] Innovative therapy for lung cancer ～Intratracheal delivery of nanomicroparticles-incorporated immune checkpoint-blocking single-chain variable fragment～ (2019)
  • Author(s): Sato T
  • Organizer: NIH/NCI Symposium on Innate Immunity & Modulation of the Host Immune Response in Honor of the Career of Dr. Dennis Klinman
  • Int'l Joint Research / Invited
• [Book] A Compendium of Chronic Obstructive Pulmonary Disease-Intratracheally Therapeutic Option for COPD: A Potential Usage of the Therapeutic Microbe for Delivering Specific Protein to the Lungs (2023)
  • Author(s): Sato T and Shimosato T
  • Total Pages: 11
  • Publisher: IntechOpen
  • ISBN: 9781803567686
• [Remarks] New Treatment Modality for Lung Diseases
  • URL: https://www.ingentaconnect.com/content/sil/impact/2019/00002019/00000003/art00022
• [Patent(Industrial Property Rights)] 医薬組成物 (2020)
  • Inventor(s): 佐藤隆、下里剛士、森谷樹、森永匡彦、佐藤祐一、小番昭宏
  • Industrial Property Rights Holder: 佐藤隆、下里剛士、森谷樹、森永匡彦、佐藤祐一、小番昭宏
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
  • Overseas
• [Funded Workshop] International workshop 2023 Inhaled Probiotic Therapy Next Era Therapeutic Modality for Lung Cancer (2023)