Cell therapy and gene therapy for heredity deafness with genetically modified mice

• Project/Area Number: 20390445
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Otorhinolaryngology
• Research Institution: Juntendo University
• Principal Investigator: IKEDA Katuhisa Juntendo University, 医学部, 教授 (70159614)
• Co-Investigator(Kenkyū-buntansha): MINOWA Osamu 理化学研究所, 上級研究員 (00181967) ; TAKESHI Kusunoki 順天堂大学, 医学部, 准教授 (30248025) ; FURUKAWA Masayuki 順天堂大学, 医学部, 准教授 (20359524) ; YOKOI Hidenori 順天堂大学, 医学部, 准教授 (80317487) ; KOIKE Takuji 電気通信大学, 電気通信学部, 准教授 (10282097) ; KAMIYA Kazusaku 順天堂大学, 医学部, 講師 (10374159) ; KAKEHATA Seiji 順天堂大学 (90261619)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥19,110,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥4,410,000); Fiscal Year 2010: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2009: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2008: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
• Keywords: GJB2 / 難聴 / ノックアウトマウス / コルチ器 / 内耳 / 蝸牛 / 内リンパ / 半規管 / アデノウイルスベクター / アデノ随伴ウイルスベクター / 聴性脳幹反応 / GJB2遺伝子 / 遺伝子導入 / ウイルスベクター / アデノウイル / アデノ随伴ウイル

Research Abstract

Congenital deafness affects about 1 in 1000 children and the half of them have genetic background such as Connexin26 gene mutation. The strategy to rescue such heredity deafness has not been developed yet. Recently, a number of clinical studies for cell therapy have been reported and clinically used for several intractable diseases. Inner ear cell therapy for sensorineural hearing loss also has been studied using some laboratory animals, although the successful reports for the hearing recovery accompanied with supplementation of the normal functional cells followed by tissue repair, recovery of the cellular/molecular functions were still few. Previously, we developed a novel animal model for acute sensorineural hearing loss due to fibrocyte dysfunction and performed cell therapy with bone marrow mesenchymal stem cells (MSC) as supplementation of cochlear fibrocytes functioning for cochlear ion transport. MSC has been known to have little risk for carcinogenesis compared with embryonic stem (ES) cell and induced pluripotent stem (iPS) cell. We injected MSC into the lateral semicircular canal and a number of these stem cells were then detected in the injured area in the lateral wall. The transplanted animals showed a significantly higher hearing recovery ratio than controls. We analyzed the machinery of this stem cell induction to the targeted site in cochlea and found that monocyte chemotactic protein 1 (MCP1) and chemokine (C-C motif) receptor 2 (CCR2) played important roles for this cell induction. To enhance the MSC induction in cochlea, we developed a novel transplant strategy by induction of MCP1 expression in host cochlear tissue and forced expression of CCR2 in MSC. Furthermore, we developed a novel mouse model for Connexin26 mutation as most frequent heredity deafness in human. With these animal models, the developed stem cells and the novel strategy to enhance the stem cell induction, we examined to establish an efficient stem cell therapy to cochlear target site followed by recovery of hearing function in heredity deafness.

Research Products

• [Journal Article] Bacteriology of recurrent exacerbation of postoperative course in chronic rhinosinusitis in relation to asthma (2011)
  • Author(s): Ikeda K, Yokoi H, Kusunoki T, Saitoh T, Yao T, Kase K, Minekawa A, Inoshita A, Kawano K
  • Journal Title: Auris Nasus Larynx(Epub ahead of print) Pages: 1312-1319
  • Peer Reviewed
• [Journal Article] Bacteriology of recurrent exacerbation of postoperative course in chronic rhinosinusitis in relation to asthma (2011)
  • Author(s): Katsuhisa IKEDA
  • Journal Title: Auris Nasus Larynx Volume: (Epub ahead of print) Pages: 1312-1319
  • Peer Reviewed
• [Journal Article] Relationship between olfactory acuity and peak expiratory flow during postoperative follow-up in chronic rhinosinusitis associated with asthma (2010)
  • Author(s): Ikeda K, Yokoi H, Kusunoki T, Saitoh T, Yao T, Kase K, Minekawa A, Inoshita A, Kawano K
  • Journal Title: Ann Otol Rhinol Laryngol 119(11) Pages: 576-581
  • Peer Reviewed
• [Journal Article] Vestibular function of patients with profound deafness related to GJB2 mutation. (2010)
  • Author(s): Kasai M, Hayashi C, Iizuka T, Inoshita A, Kamiya K, Okada H, Nakajima Y, Kaga K, Ikeda K
  • Journal Title: Acta Otolaryngol. 130(9) Pages: 990-5
  • Peer Reviewed
• [Journal Article] Relationship between olfactory acuity and peak expiratory flow during postoperative follow-up in chronic rhinosinusitis associated with asthma (2010)
  • Author(s): Katsuhisa IKEDA
  • Journal Title: Ann Otol Rhinol Laryngol Volume: 119(11) Pages: 576-581
  • Peer Reviewed
• [Journal Article] Cochlear outer hair cells in a dominant-negative connexin26 mutant mouse preserve non-linear capacitance in spite of impaired distortion product otoacoustic emission. (2009)
  • Author(s): Minekawa A, Abe T, Inoshita A, Iizuka T, Kakehata S, Narui Y, Koike T, Kamiya K, Okamura HO, Shinkawa H, Ikeda K
  • Journal Title: Neuroscience 164 Pages: 1312-1319
• [Journal Article] Relationship between epithelial damage or basement membrane thickness and eosinophilic infiltration in nasal polyps with chronic rhinosinusitis. (2009)
  • Author(s): Saito T, Kusunoki T, Yao T, Kawano K, Kojima Y, Miyahara K, Onoda J, Yokoi H, Ikeda K
  • Journal Title: Rhinology 47 Pages: 275-9
• [Journal Article] Development of distortion product Otoacoustic emissions in C57BL/6J mice. (2009)
  • Author(s): Narui Y, Minekawa A, Iizuka T, Furukawa M, Kusunoki T, Koike T, Ikeda K
  • Journal Title: Int J Audiol 48 Pages: 576-81
• [Journal Article] Overexpression of GLUT-1 in the invasion front is associated with depth of oral squamous cell carcinoma and prognosis. (2009)
  • Author(s): Ohba S, Fujii H, Ito S, Fujimaki M, Matsumoto F, Furukawa M, Yokoyama J, Kusunoki T, Ikeda K
  • Journal Title: Hino O J Oral Pathol Med.
• [Journal Article] Role of Interleukin-17A in the Eosinophil Accumulation and Mucosal Remodeling in Chronic Rhinosinusitis with Nasal Polyps Associated with Asthma. (2009)
  • Author(s): Saitoh T, Kusunoki T, Yao T, Kawano K, Kojima Y, Miyahara K, Onoda J, Yokoi H, Ikeda K
  • Journal Title: Int Arch Allergy Immunol 151 Pages: 8-16
• [Journal Article] Eotaxin-1, -2, and -3 immunoreactivity and protein concentration in the nasal polyps of eosinophilic chronic rhinosinusitis patients. (2009)
  • Author(s): Yao T, Kojima Y, Koyanagi A, Yokoi H, Saito T, Kawano K, Furukawa M, Kusunoki T, Ikeda K
  • Journal Title: Laryngoscope 119 Pages: 1053-9
• [Journal Article] Development of distortion product Otoacoustic emissions in C57BL/6J mice (2009)
  • Author(s): Narui Y, Minekawa A, Iizuka T, Furukawa M, Kusunoki T, Koike T, Ikeda K.
  • Journal Title: Int J Audiol 48 Pages: 576-81
  • Peer Reviewed
• [Journal Article] Cochlear outer hair cells in a dominant-negative connexin26 mutant mouse preserve non-linear capacitance in spite of impaired distortion product otoacoustic emission. (2009)
  • Author(s): Minekawa A, Abe T, Inoshita A, Iizuka T, Kakehata S, Narui Y, Koike T, Kamiya K, Okamura HO, Shinkawa H, Ikeda K.
  • Journal Title: Neuroscience 164 Pages: 1312-1319
  • Peer Reviewed
• [Journal Article] Noninvasive in vivo delivery of transgene via adeno-associated virus into supporting cells of the neonatal mouse cochlea (2008)
  • Author(s): Iizuka T, Kanzaki S, Mochizuki H, Inoshita A, Narui Y, Furukawa M, Kusunoki T, Saji M, Ogawa K, Ikeda K
  • Journal Title: Hum Gene Ther 19 Pages: 384-390
  • Peer Reviewed
• [Journal Article] Postnatal development of the organ of Corti in dominant-negative Gjb2 transgenic mice (2008)
  • Author(s): Inoshita A, Iizuka T, Okamura HO, Minekawa A, Kojima K, Furukawa M, Kusunoki T, Ikeda K
  • Journal Title: Neuroscience 156 Pages: 1039-1047
  • Peer Reviewed
• [Journal Article] Noninvasive In Vivo Delivery of Transgene via Adeno-Associated Virus into Supporting Cells of the Neonatal Mouse Cochlea. (2008)
  • Author(s): Takashi IIZUKA
  • Journal Title: Human Gene Therapy 19(4) Pages: 384-390
  • Peer Reviewed
• [Journal Article] Postnatal development of the organ of Corti in dominant-negative Gjb2 transgenic mice. (2008)
  • Author(s): Ayako INOSHITA
  • Journal Title: Neuroscience 156(4) Pages: 1039-1047
  • Peer Reviewed
• [Presentation] 欧州分子生物学会(EMBO meeting) (2010)
  • Author(s): 神谷和作、池田勝久
  • Organizer: Cochlear Gap-Junction plaque is disrupted by dominant-negative Connexin26 mutation.
  • Place of Presentation: スペイン・バルセロナ
  • Year and Date: 2010-09-04
• [Presentation] Cochlear Gap-Junction plaque is disrupted by dominant-negative Connexin26 mutation (2010)
  • Author(s): K Ikeda
  • Organizer: Inner Ear Biology
  • Place of Presentation: チェコ共和国・プラハ
  • Year and Date: 2010-09-01
• [Presentation] Gene therapy in GJB2 KO mice (2010)
  • Author(s): Iizuka T, Ikeda K
  • Organizer: Association of research in Otolaryngology
  • Place of Presentation: Anaheim
  • Year and Date: 2010-02-08
• [Presentation] コネキシン26優性阻害変異によるコルチ器周囲細胞におけるギャップ結合プラークの形成変化 (2009)
  • Author(s): 神谷和作、池田勝久
  • Organizer: 日本耳科学会学術集会
  • Place of Presentation: 東京
• [Remarks] ホームページ等