Regulation of Vascular Remodeling by Gene Transfer and Its Clinical Application : Exploration of Novel Gene Transfer Vector and Gene Therapy for Atherosclerosis, Angiogenic Diseases and Organ Infarction

基因转移调控血管重塑及其临床应用：新型基因转移载体的探索以及动脉粥样硬化、血管生成疾病和器官梗塞的基因治疗

• 批准号: 11557017
• 负责人: SUEISHI Katsuo
• 金额: $ 7.62万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557017/
• 关键词: gene transfer; SeV; VEGF; FGF-2; MCP-1; safety; therapeutic angiogenesis; gene therapy; SeV; SeVベクター; 動脈硬化; 血管新生病; 移植

The major results obtained in the last year (from Apr., 2000 to Mar., 2001) were as follows : 1. Investigation on biological properties of Sendai virus vector (SeV) : 1) the in vitro and in vivo efficiencies of transduction and expression of beta-gal., luciferase, ecNOS, VEGF, FGF-2 and other genes using SeV were almost the same as those by Adeno virus vector, and SeV-mediated gene transfer was characterized by the fact that its brief exposure to cells in vitro and in vivo, less than 10 min., was enough for the peak gene expression. 2) Intaluminal injection of reporter gene-SeV could label not only ECs but also medial SMCs, but the neointima ocurred in human varicose veins hampered the transfection of medial SMCs.2. In order to confirm the safety of SeV, we examined not only the systemic distribution of reporter genes and sendai virus genom but also the degree of inflammatory reaction including serum/plasma levels of such cytokines as IL-1, 6, 8, TNF-alpha and others. We are now prepar … More ing the format of "Therapeutical application of FGF-2 gene injection to patients with critical limb ischemia using SeV" for getting the approval in the "Ethical Committee of Graduate School of medical Sciences, Kyushu University.3. Molecular mechanism of angiogenesis in ischemic tissues : 1) Effect of intramuscular injection of VEGF and FGF-2 genes-SeV on development of collateral vessels in ischemic limb mouse model : injection of FGF-2 gene could induce endogeneous VEGF overexpression, and markedly improve blood perfusion in the ischemic limb by enhanced angiogenesis. However, VEGF gene injection worsened blood perfusion in ischemic limb. 2) the final and target angiogenic molecule for accelerating the development of collateral vessels in ischemic tissues was VEGF, but the harmonized collaboration of other angiogenic factors including FGF-2 was necessary for developing effective collateral circulation.4. In rat pulmonary hypertension induced by monocrotaline, interstitial remodeling by function of infiltrating macrophages through MCP-1 was revealed to be most important in development of pulmonary hypertension, and intra- muscular infection of 7ND MCP-1 gene was effective for its prevention.5. Pigmentary retinopathy was revealed to be an angiogenic disease by the development of animal models of retroretinal angiogenesis by overexpression of VEGF using SeV. Less

去年取得的主要成果（从4月，2000年至3月，2001年）如下：1。仙台病毒载体（SeV）生物学特性的研究：1）β-gal.的体内外转导效率和表达效率，使用SeV的荧光素酶、ecNOS、VEGF、FGF-2和其他基因与腺病毒载体的几乎相同，SeV介导的基因转移的特征在于其在体外和体内短暂暴露于细胞，少于10分钟，足以让基因表达达到峰值。2)腔内注射报告基因SeV不仅可以标记内皮细胞，还可以标记中膜SMC，但曲张静脉内的新生内膜阻碍了中膜SMC的转染.为了确认SeV的安全性，我们不仅检查了报告基因和仙台病毒基因组的全身分布，而且还检查了炎症反应的程度，包括诸如IL-1、6、8、TNF-α等细胞因子的血清/血浆水平。我们现在准备 ...更多信息 将“使用SeV的FGF-2基因注射在严重肢体缺血患者中的治疗应用”的格式提交给“九州大学医学科学研究生院伦理委员会”以获得批准。缺血组织血管生成的分子机制：1）肌肉注射VEGF和FGF-2基因-SeV对缺血肢体侧支血管形成的影响：注射FGF-2基因可诱导内源性VEGF过度表达，通过增强血管生成明显改善缺血肢体的血液灌注。而VEGF基因注射使缺血肢体血流灌注恶化。2)促进缺血组织侧支血管形成的最终靶向血管生成分子是VEGF，但包括FGF-2在内的其他血管生成因子的协调合作对于形成有效的侧支循环是必要的.在野百合碱诱导的大鼠肺动脉高压模型中，MCP-1介导的巨噬细胞浸润导致的肺间质重构在肺动脉高压的发生中起重要作用，肌肉注射7 ND MCP-1基因可有效预防肺间质重构.通过使用SeV过表达VEGF来开发视网膜后血管生成的动物模型，揭示了色素性视网膜病变是一种血管生成性疾病。少

项目成果

畑快右: "血管研究の最前線に迫る"羊土社. 9 (2000)

羽田义介：“走近血管研究的前沿”Yodosha 9 (2000)。

Sakamoto T,et al: "Target gene transfer of tissue plasminogen activator to cornea by electric pulse inhibits intracameral fibrin formation and cornal cloudiness."Hum Gene Ther. 10. 2551-2557 (1999)

Sakamoto T 等人：“通过电脉冲将组织纤溶酶原激活剂的目标基因转移至角膜，抑制前房内纤维蛋白形成和角膜混浊。”Hum Gene Ther。

Masaki I, et al: "Recombinant Sendai virus-mediated gene transfer to vasculature : a new class of efficient gene transfer vector to the vascular system."FASEB J. (in press).

Masaki I 等人：“重组仙台病毒介导的基因转移到脉管系统：一类新的有效基因转移载体到血管系统。”FASEB J.（出版中）。

Sakamoto T, et al: "Target gene transfer of tissue plasminogen activator to cornea by electric pulse inhibits intracameral fibrin formation and corneal cloudiness."Hum Gene Ther. 10. 2551-2557 (1999)

Sakamoto T 等人：“通过电脉冲将组织纤溶酶原激活剂的目标基因转移至角膜，抑制前房内纤维蛋白形成和角膜混浊。”Hum Gene Ther。

Yonemitsu Y: "Methods in Molecular Medicine, Vascular Disease : Molecular Biology and Gene Therapy Protocols"Humana Press Inc. 12 (1999)

Yonemitsu Y：“分子医学、血管疾病的方法：分子生物学和基因治疗方案”Humana Press Inc. 12 (1999)