Improving fibrin-based bioartificial arteries by prolonging ERK activation

描述(申请人提供)：在全国范围内，冠状动脉疾病是一个巨大的医疗负担。目前治疗冠状动脉疾病的方法对患者来说存在多种风险，包括再狭窄、血栓形成、感染和其他移植物疾病。一种完全生物的人工动脉移植物将绕过这些问题，并改善冠心病患者的预后。基于纤维蛋白的组织工程学已经取得了重大进展，但迄今还没有制造出机械强度足够的生物人工动脉，可以无风险地植入。需要一种新的策略来利用信号通路来实现细胞刺激。这项工作的假设是：在体外培养过程中延长ERK信号将通过刺激I型胶原转录和增加胶原含量来提高基于纤维蛋白的生物人工动脉的机械强度。通过控制细胞外信号调节激酶(ERK)信号转导，该项目将提高生物人工动脉的胶原蛋白含量，最终提高其机械强度。拟议工作的具体目标如下：#1.确定ERK活性对于产生机械强度较强的生物人工动脉是必要的。#2.通过抑制负反馈通路促进生物人工动脉中ERK的延长激活。#3.通过机械刺激促进生物人工动脉中ERK的延长激活。对ERK信号操纵的反应的主要读数将是I型胶原转录，使用荧光素酶报告，胶原含量，使用生化分析，以及机械强度，使用机械测试系统。明尼苏达大学专家在组织工程和生物力学方面的重要培训将是该奖学金的关键目标。预计该项目将生产出能够承受生理血压的生物人造动脉。

DESCRIPTION (provided by applicant): Nationally, coronary artery disease is a tremendous health care burden. Current therapies for coronary artery disease suffer from multiple risks to the patient including restenosis, thrombosis, infection, and other graft disease. A completely biological bioartificial artery graft would circumvent these issues and improve the outcome for sufferers of coronary artery disease. Fibrin-based tissue engineering has already shown significant progress but as yet has not produced a bioartificial artery with sufficient mechanical strength for implantation without risk. A new strategy is needed to exploit signaling pathways for cellular stimulation. The hypothesis for the proposed work is that: Prolonged ERK signaling during in vitro culture will improve the mechanical strength of fibrin-based bioartificial arteries, via stimulation of type I collagen transcription and increased collagen content. By manipulating extracellular signal-regulated kinase (ERK) signaling by the cells seeded in tubular fibrin- based constructs, this project will improve the collagen content of bioartificial arteries and ultimately their mechanical strength. The specific aims of the proposed work are as follows: #1. Establish that ERK activity is necessary for the production of mechanically strong bioartificial arteries. #2. Promote prolonged ERK activation in bioartificial arteries by inhibiting negative feedback pathways. #3. Promote prolonged ERK activation in bioartificial arteries by mechanical stimulation. The primary readouts for the response to ERK signal manipulation will be type I collagen transcription, using a luciferase reporter, collagen content, using a biochemical assay, and mechanical strength, using mechanical testing systems. Significant training in tissue engineering and biomechanics from experts at the University of Minnesota will be a key goal for this fellowship. It is expected that this project will produce bioartificial arteries that can withstand physiological blood pressure.