Improving fibrin-based bioartificial arteries by prolonging ERK activation

描述（由申请人提供）：在全国范围内，冠状动脉疾病是一个巨大的卫生保健负担。目前用于冠状动脉疾病的疗法对患者具有多种风险，包括再狭窄、血栓形成、感染和其它移植物疾病。一个完全生物的生物人工动脉移植物将避免这些问题，并改善冠状动脉疾病患者的结果。基于纤维蛋白的组织工程已经显示出显著的进展，但迄今为止还没有产生具有足够的机械强度以用于无风险植入的生物人工动脉。需要一种新的策略来利用细胞刺激的信号通路。提出的工作的假设是：在体外培养期间延长ERK信号传导将通过刺激I型胶原转录和增加胶原含量来改善基于纤维蛋白的生物人工动脉的机械强度。通过操纵细胞外信号调节激酶（ERK）信号传导的细胞接种在管状纤维蛋白为基础的结构，该项目将提高生物人工动脉的胶原蛋白含量，并最终提高其机械强度。具体工作目标如下：#1。确定ERK活性是生产机械强度强的生物人工动脉所必需的。#2.通过抑制负反馈通路促进生物人工动脉中ERK的长期激活。#3.通过机械刺激促进生物人工动脉中ERK的长期激活。对ERK信号操纵的响应的主要读数将是I型胶原蛋白转录（使用荧光素酶报告基因）、胶原蛋白含量（使用生物化学测定）和机械强度（使用机械测试系统）。来自明尼苏达大学的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一个关键目标。预计该项目将生产出能够承受生理血压的生物人工动脉。 公共卫生相关性：这项拟议中的研究将加速生产完全生物可植入的生物人工动脉，这是目前正确治疗冠状动脉疾病的需要。通过在动脉发育期间操纵细胞信号传导，胶原蛋白含量将增加，导致机械强度增强。

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. PUBLIC HEALTH RELEVANCE: The proposed research will accelerate the production of a completely biological implantable bioartificial artery, which is a current need for proper treatment of coronary artery disease. By manipulating cellular signaling during artery development, collagen content will be increased, leading to enhanced mechanical strength.