脳神経病変を標的としたリソゾーム病の新規治療法（遺伝子治療）の開発

2013 Fiscal Year Research-status Report

• Project/Area Number: 24591529
• Research Institution: Nippon Medical School
• Principal Investigator: 三宅 紀子 日本医科大学, 医学部, その他 (00421206)
• Co-Investigator(Kenkyū-buntansha): 三宅 弘一 日本医科大学, 医学部, 准教授 (90267211)
• Keywords: 異染性白質ジストロフィー / 新生児治療 / AAV vector / 遺伝子治療

Research Abstract

本研究では脳全体の広範な神経変性を伴う、MLDをモデルとして、①新生児期の治療法の確立、及びBBBを効率よく通過するための画期的な方法による②成体の神経症状の新規治療法の確立を行っていくとともに、臨床応用を念頭に安全性の検討も行い、臨床プロトコールの作製を行う。
1. 新生児期治療法の確立：ASA発現治療用AAVベクター（9型）を新生児期のMLD病のモデルマウスに静脈内投与し、脳神経組織への導入効率、ASA発現効率を解析すると共に、蓄積物質であるスルファチドの定量、行動実験などにて治療効果判定を行う。
2. 成体の神経症状の新規治療法の確立：治療用AAVベクターの改良（BBBを通過可能な血清型、scAAV）及び治療タンパクの改変（ASA-Tatキメラ蛋白質の作製）を行い、脳神経組織治療に最良のベクターの選択を行う。そのベクターを用いて成体MLD病のモデルマウスに静脈内投与、髄腔内投与にて治療効果判定を行っていく。
また、これらの治療モデルマウスを長期に観察し、副作用の検討も行い、これらの結果を基に臨床応用を目的としたプロトコールを作製する。
当該年度に実施した内容は、治療用self-complementary AAV (scAAV)ベクターを作製し成体のMLDマウスに静脈注射を行い、治療効果の判定を行った。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

成体の神経症状の新規治療法の確立
当該年度は、治療用self-complementary AAV (scAAV)ベクターを作製、成体のMLDマウスに静脈注射を行い、治療効果の判定を行った。第一段階として、行動実験による治療効果の判定を行ったが、未治療群に比べ優位に改善効果を認められた。

Strategy for Future Research Activity

前年度から長期観察をしている新生児期治療の毒性試験の解析を行う。また成体マウスへ治療し行動実験にて効果を認められたマウスの解析を行う。

Expenditure Plans for the Next FY Research Funding

見積もり安い金額で納品されたため差額が生じた。
次年度に使用したい。

Research Products

• [Journal Article] Long-term correction of biochemical and neurological abnormalities in MLD mice model by neonatal systemic injection of an AAV serotype 9 vector. (2014)
  • Author(s): Miyake N, Miyake K, Asakawa N, Yamamoto M, Shimada T.
  • Journal Title: Gene Ther. Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1038/gt.2014.17
  • Peer Reviewed
• [Journal Article] Inhibition of S100A6 induces graft versus leukemia effects in MLL/AF4-positive ALL in human PBMC-SCID mice. (2014)
  • Author(s): Tamai H, Miyake K, Yamaguchi H, Dan K, Inokuchi K.
  • Journal Title: Bone Marrow Transplant. Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1038/bmt.2014.18.
  • Peer Reviewed
• [Journal Article] siRNA Knockdown of Tissue Inhibitor of Metalloproteinase-1 in Keloid Fibroblasts Leads to Degradation of Collagen Type I. (2014)
  • Author(s): Aoki M, Miyake K, Ogawa R, Dohi T, Akaishi S, Hyakusoku H, Shimada T
  • Journal Title: J Invest Dermatol. Volume: 134 Pages: 818-826
  • DOI: 10.1038/jid.2013.396
  • Peer Reviewed
• [Journal Article] Mutant ataxin-3 with an abnormally expanded polyglutamine chain disrupts dendritic development and metabotropic glutamate receptor signaling in mouse cerebellar purkinje cells. (2014)
  • Author(s): Konno A, Shuvaev AN, Miyake N, Miyake K, Iizuka A, Matsuura S, Huda F, Nakamura K, Yanagi S, Shimada T, Hirai H.
  • Journal Title: Cerebellum. Volume: 13 Pages: 29-41
  • DOI: 10.1007/s12311-013-0516-5
  • Peer Reviewed
• [Journal Article] Cyclical mechanical stretch enhances degranulation and IL-4 secretion in RBL-2H3 mast cells. (2014)
  • Author(s): Komiyama H, Miyake K, Asai K, Mizuno K, Shimada T.
  • Journal Title: Cell Biochem Funct. Volume: 32 Pages: 70-76
  • DOI: 10.1002/cbf.2973
  • Peer Reviewed
• [Journal Article] miR-7a alleviates the maintenance of neuropathic pain through regulation of neuronal excitability. (2013)
  • Author(s): Sakai A, Saitow F, Miyake N, Miyake K, Shimada T, Suzuki H.
  • Journal Title: Brain. Volume: 136 Pages: 2738-2750
  • DOI: 10.1093/brain/awt191
  • Peer Reviewed
• [Journal Article] Role of survivin in acute lung injury: epithelial cells of mice and humans. (2013)
  • Author(s): Terasaki Y, Terasaki M, Urushiyama H, Nagasaka S, Takahashi M, Kunugi S, Ishikawa A, Wakamatsu K, Kuwahara N, Miyake K, Fukuda Y.
  • Journal Title: Lab Invest. Volume: 93 Pages: 1147-1163
  • DOI: 10.1038/labinvest.2013.103.
  • Peer Reviewed
• [Journal Article] Direct comparison of administration routes for AAV8-mediated ocular gene therapy. (2013)
  • Author(s): Igarashi T, Miyake K, Asakawa N, Miyake N, Shimada T, Takahashi H.
  • Journal Title: Curr Eye Res. Volume: 38 Pages: 569-577
  • DOI: 10.3109/02713683.2013.779720.
  • Peer Reviewed
• [Presentation] Minimally invasive gene therapy of MLD model mice by intrathecal and intravenous injection of AAV9 vector. (2014)
  • Author(s): Noriko Miyake, Koichi Miyake, and Takashi Shimada
  • Organizer: 10th WORLDSymposium™ 2014
  • Place of Presentation: San Diego,CA
  • Year and Date: 20140210-20140213
• [Presentation] Tolerance induction to arylsulfatase A by AAV mediated neonatal gene transfer into MLD model mice. (2013)
  • Author(s): Miyake N, Miyake K, Yamamoto M and Shimada T.
  • Organizer: The 19th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: Okayama
  • Year and Date: 20130704-20130706
• [Presentation] siRNA Targeting Vascular Endothelial Growth Factor by adeno-associated vector (type 8) efficiently inhibits Neovascularization in a Murine Choroidal Neovascularization Model. (2013)
  • Author(s): Igarashi T, Miyake N, Iijima O, Yaguchi C, Shimada T, Takahashi H and Miyake K.
  • Organizer: The 19th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: Okayama
  • Year and Date: 20130704-20130706
• [Presentation] Disruption of metabotropic glutamate receptor signaling and rescue through intravascular administration of AAV9 in a mouse model of spinocerebellar ataxia type 3. (2013)
  • Author(s): Konno A, Shuavaev AN, Miyake N, Miyake K, Yanagi S, Shimada T, Hirai H.
  • Organizer: The 19th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: Okayama
  • Year and Date: 20130704-20130706
• [Presentation] Robust and cell type-specific transgene expression by applying tetracycline-controlled system to AAV9. (2013)
  • Author(s): Yamaguchi T, Konno A, Miyake N, Miyake K, Shimada T, Hirai H.
  • Organizer: The 19th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: Okayama
  • Year and Date: 20130704-20130706
• [Presentation] Long−Term Secretion of Arylsulfatase A without Antibody Formation after AAV Mediated Neonatal Gene Transfer into MLD Model Mice. (2013)
  • Author(s): Miyake N, Miyake K, Yamamoto M and Shimada T.
  • Organizer: 16th Annual Meeting of the American Society of Gene & Cell Therapy.
  • Place of Presentation: Salt Lake City, Utah
  • Year and Date: 20130515-20130518
• [Presentation] Adeno−Associated Vector (Type 8)−Mediated Expression of siRNA Targeting Vascular Endothelial Growth Factor Efficiently Inhibits Neovascularization in a Murine Choroidal Neovascularization Model. (2013)
  • Author(s): Igarashi T, Miyake K, Miyake N, Iijima O, Yaguchi C, Shimada T and Takahashi H.
  • Organizer: 16th Annual Meeting of the American Society of Gene & Cell Therapy.
  • Place of Presentation: Salt Lake City, Utah
  • Year and Date: 20130515-20130518
• [Book] Novel gene therapy approaches (2013)
  • Author(s): Miyake K, Shimada T.
  • Total Pages: 440
  • Publisher: InTech