Analysis of biological function of tenascin-C in progression of heart failure and its clinical application

Tenascin-C在心力衰竭进展中的生物学功能分析及其临床应用

• 批准号: 10670644
• 负责人: IMAI Hiroshi
• 金额: $ 2.43万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670644/
• 关键词: Extracellular matrix; tenascin; myofibroblast; knockout mouse; myocardial infarction; myocarditis; 筋繊維芽細胞; 心疾患

In order to study the biological function, recombinant protein of tenascin-C was added to the cell cultures.1) Tenascin-C, collaborating with fibronectin, accelerates the initial attachment of cardiomyocytes isolated from adult rat to laminin, however, tenascin-C is not involved in the formation of costameric attachment. 2) Tenascin-C induces the differentiation of fibroblasts to myofibroblasts. Secondary, to clarify the role of tenascin-C if myocardial tissue in vivo, we examined the wound healing of a mouse myocardial infarction model using tenascin-C knockout mouse. In tenascin-C null mutant, the number of smooth-muscle actin-positive cells was delayed to compare to that of normal mouse, however, healing proceeded normally. Tenascin-X, another member of tenascin family was ubiquitously expressed in normal heart, and slightly unregulated after myocardial injury at later stage. Our findings did not suggested that tenascin-X compensates for the loss of tenascin-C. Finally, to examine the possibility of the clinical application anti tenascin-C, we have established a new autoimmune mouse model. We also raised a mouse monoclonal antibody, which recognizes mouse tenascin-C by immunization of a tenascin-C knock out mouse. Immunoshitological study showed that tenascin-C expressed at very early stage of active phase of myocarditis. Our data demonstrate that tenascin-C is a useful marker for histological diagnosis to evaluate disease activity of myocarditis.

为了研究其生物学功能，将重组蛋白tenascin-C添加到细胞培养物中。1) Tenascin-C与纤连蛋白协同作用，加速成年大鼠分离的心肌细胞与层粘连蛋白的初始附着，但tenascin-C不参与肋节附着的形成。 2) Tenascin-C诱导成纤维细胞分化为肌成纤维细胞。其次，为了阐明生腱蛋白-C 在体内心肌组织中的作用，我们使用生腱蛋白-C 敲除小鼠检查了小鼠心肌梗死模型的伤口愈合情况。在生腱蛋白-C无效突变体中，与正常小鼠相比，平滑肌肌动蛋白阳性细胞的数量延迟，但愈合过程正常进行。 Tenascin-X是腱蛋白家族的另一成员，在正常心脏中广泛表达，在心肌损伤后期稍微不受调节。我们的研究结果并未表明生腱蛋白-X 可以补偿生腱蛋白-C 的损失。最后，为了检验抗生腱蛋白-C临床应用的可能性，我们建立了一种新的自身免疫小鼠模型。我们还制备了一种小鼠单克隆抗体，它通过免疫腱蛋白-C 敲除小鼠来识别小鼠腱蛋白-C。免疫组织学研究表明，tenascin-C在心肌炎活动期的早期阶段表达。我们的数据表明生腱蛋白-C 是组织学诊断评估心肌炎疾病活动性的有用标记物。

项目成果

Imanaka-Yoshida, K., et al.: "Vinculin, Talin, Integrin alpha6betal and laminin can serve as components of attachment complex mediating contraction force transmission from cardiomyocytes to extracellular matrix"Cell Motil Cytoskeleton. 42. 1-11 (1999)

Imanaka-Yoshida, K. 等人：“Vinculin、Talin、整合素 α6β1 和层粘连蛋白可以作为附着复合物的成分，介导从心肌细胞到细胞外基质的收缩力传递”Cell Motil 细胞骨架。

Imanaka-Yoshida, K., et al.: "N-cadherin is required for the differentiation and initial myofibrillogenesis of chick cardiomyocytes"Cell Motil Cytoskeleton. 39. 52-62 (1998)

Imanaka-Yoshida, K. 等人：“鸡心肌细胞的分化和初始肌原纤维形成需要 N-钙粘蛋白”Cell Motil Cytosculpture。

Imanaka-Yoshida,K.,et al.: "Vinculin,Talin,Integrin alpha6betal and laminin can serve as components of attachment complex mediating contraction force transmission from cardiomyocytes to extracellular matrix"Cell Motil Cytoskeleton. 42. 1-11 (1999)

Imanaka-Yoshida, K. 等人：“Vinculin、Talin、整合素 α6β1 和层粘连蛋白可以作为附着复合物的成分，介导从心肌细胞到细胞外基质的收缩力传递”Cell Motil 细胞骨架。

Yoshida, T., et al.: "Low cytoplasmic pH causes fragmentation and dispersal of the Golgi apparatus in human"Int J Exp Pathol. 80. 51-57 (1999)

Yoshida, T., et al.：“低细胞质 p​​H 值导致人类高尔基体碎裂和分散”Int J Exp Pathol。

Imaoka-Yoshida, K., et al.: "N-cadherin is required for the differentiation and initial myofibrillogenesis of chick cardiomyocytes"Cell Motil Cytoskeleton. 39. 52-62 (1998)

Imaoka-Yoshida, K. 等人：“鸡心肌细胞的分化和初始肌原纤维生成需要 N-钙粘蛋白”Cell Motil Cytosculpture。