The development of new therapy for pulmonary fibrosis by using exogenous protein delivery system mimicked biological mechanism

• Project/Area Number: 15K15322
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Respiratory organ internal medicine
• Research Institution: Nagoya University
• Principal Investigator: HASHIMOTO NAOZUMI 名古屋大学, 医学部附属病院, 講師 (30378020)
• Co-Investigator(Kenkyū-buntansha): 長谷川 好規 名古屋大学, 医学系研究科, 教授 (20270986)
• Research Collaborator: Miyazaki Shinichi 名古屋大学, 大学院医学系研究科
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 分子細胞尾呼吸器学 / 線維芽細胞 / 細胞内導入デリバリーシステム / PTEN / 肺線維症 / 上皮細胞 / 微小環境 / 上皮間葉系移行

Outline of Final Research Achievements

We established the cells with the GFP-fused domain to induce secretion of PTEN (N173GFP). The additional kozak sequence to an alternative translation start site of PTEN improved the expression level of N173GFP. Although intracellular expression of N173GFP was detected, N151GFP, the secreted type of N173GFP, could not be detected in the culture media. To directly treat epithelial cells with exogenous PTEN, we established the cells which produce the GFP-fused secreted alternative domain of PTEN (N151GFP). When exogenous administration of GFPN151 to epithelial cells was performed in vitro, immunofluorescent evaluation could not show GFP expression in the cells. By not only the gene modification of alternative translation start site of PTEN to improve expression rate but also the determination of appropriate condition to improve recovery rate of secreted PTEN protein, we establish the new therapeutic strategy to regulate tissue microenvironment.

Research Products

• [Journal Article] Clinical impact of the lower limit of normal of FEV1/FVC on survival in lung cancer patients undergoing thoracic surgery (2016)
  • Author(s): Matsuzaki A, Hashimoto N, Okachi S, Taniguchi T, Kawaguchi K, Fukui T, Wakai K, Yokoi K, Hasegawa Y.
  • Journal Title: Respir Investig Volume: 54 Pages: 184-192
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Impact of thin-section computed tomography-determined combined pulmonary fibrosis and emphysema on outcomes among patients with resected lung cancer (2016)
  • Author(s): Hashimoto N, Iwano S, Kawaguchi K, Fukui T, Fukumoto K, Nakamura S, Mori S, Sakamoto K, Wakai K, Yokoi K, Hasegawa Y.
  • Journal Title: Ann Thorac Surg Volume: 102 Pages: 440-447
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Hypoxia-induced modulation of PTEN activity and EMT phenotypes in lung cancers (2016)
  • Author(s): Kohnoh T, Hashimoto N, Ando A, Sakamoto K, Miyazaki S, Aoyama D, Kusunose M, Kimura M, Omote N, Imaizumi K, Kawabe T, Hasegawa Y.
  • Journal Title: Cancer Cell International Volume: 16 Pages: 33-42
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Factors Affecting the Diagnostic Yield of Transbronchial Biopsy Using Endobronchial Ultrasonography with a Guide Sheath in Peripheral Lung Cancer (2016)
  • Author(s): Okachi S, Imai N, Imaizumi K, Iwano S, Ando M, Hase T, Aso H, Morise M, Wakahara K, Ito S, Hashimoto N, Sato M, Kondo M, Hasegawa Y.
  • Journal Title: Intern Med Volume: 55 Pages: 1705-1712
  • NAID: 130005161019
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Antineutrophil Cytoplasmic Antibody-associated Vasculitis Superimposed on Infection-related Glomerulonephritis Secondary to Pulmonary Mycobacterium avium Complex Infection (2016)
  • Author(s): Asano S, Mizuno S, Okachi S, Aso H, Wakahara K, Hashimoto N, Ito S, Kozaki Y, Katsuno T, Maruyama S, Hasegawa Y.
  • Journal Title: Intern Med Volume: 55 Pages: 2439-2445
  • NAID: 130005262546
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Risk stratification by the lower limit of normal of FEV1/FVC for postoperative outcomes in patients with COPD undergoing thoracic surgery. (2015)
  • Author(s): Osuka S, Hashimoto N, Sakamoto K, Wakai K, Yokoi K, Hasegawa Y.
  • Journal Title: Respir Investig Volume: 53 Issue: 3 Pages: 117-123
  • DOI: 10.1016/j.resinv.2015.01.005
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Evaluation of emphysema using three-dimensional computed tomography: association with postoperative complications in lung cancer patients. (2015)
  • Author(s): Kawakami K, Iwano S, Hashimoto N, Hasegawa Y, Naganawa S.
  • Journal Title: Nagoya J Med Sci Volume: 77 Pages: 113-122
  • NAID: 120005549557
  • Peer Reviewed
• [Journal Article] Factors secreted from dental pulp stem cells show multifaceted benefits for treating acute respiratory distress syndrome in mice. (2015)
  • Author(s): Wakayama H, Hashimoto N, Matsushita Y, Matsubara K, Yamamoto N, Hasegawa Y, Ueda M, Yamamoto A
  • Journal Title: Cytotherapy Volume: 17 Issue: 8 Pages: 1119-1129
  • DOI: 10.1016/j.jcyt.2015.04.009
  • Peer Reviewed
• [Journal Article] Bronchus sign on thin-section computed tomography is a powerful predictive factor for successful transbronchial biopsy using endobronchial ultrasound with a guide sheath for small peripheral lung lesions: a retrospective observational study. (2015)
  • Author(s): Minezawa T, Okamura T, Yatsuya H, Yamamoto N, Morikawa S, Yamaguchi T, Morishita M, Niwa Y, Takeyama T, Mieno Y, Hoshino T, Uozu S, Goto Y, Hayashi M, Isogai S, Matsuo M, Nakanishi T, Hashimoto N, Okazawa M, Imaizumi K.
  • Journal Title: BMC Medical Imaging Volume: 21 Pages: 21-21
  • Peer Reviewed
• [Journal Article] Direct regulation of transforming growth factor β-induced epithelial-mesenchymal transition by the protein phosphatase activity of unphosphorylated PTEN in lung cancer cells. (2015)
  • Author(s): Kusunose M, Hashimoto N, Kimura M, Ogata R, Aoyama D, Sakamoto K, Miyazaki S, Ando A, Omote N, Imaizumi K, Kawabe T, Hasegawa Y.
  • Journal Title: Cancer Science Volume: 106 Issue: 12 Pages: 1693-704
  • DOI: 10.1111/cas.12831
  • Peer Reviewed / Open Access
• [Presentation] The PTEN protein, not lipid, phosphatase activity negatively regulate TGFβ-induced EMT in lung alveolar cells. (2015)
  • Author(s): Hashimoto N, Aoyama D, Hasegawa Y
  • Organizer: The 2015 American Thoracic Society (ATS) International Conference
  • Place of Presentation: デンバー、米国
  • Year and Date: 2015-05-17
  • Int'l Joint Research