Cues for cell-mediated regeneration of elastin matrix to stabilize aortic aneurys

细胞介导的弹性蛋白基质再生稳定主动脉瘤的线索

• 批准号: 7834746
• 负责人: ANAND RAMAMURTHI
• 金额: $ 3.44万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7834746
• 关键词: Abdominal Aortic Aneurysm; Address; Adult; Allografting; Aneurysm; Aortic Aneurysm; Architecture; Attenuated; Biological; Blood Vessels; Blood flow; Bypass; Caliber; Cell Culture Techniques; Cells; Cessation of life; Charge; Copper; Cues; Deposition; Development; Disease; Dose; Early treatment; Elastases; Elastic Fiber; Elastin; Elastin Fiber; Enzymes; Erythrocytes; Exposure to; Extracellular Matrix; Failure; Fiber; Future; Glycosaminoglycans; Goals; Growth Factor; Healed; Homeostasis; Hyaluronan; In Situ; In Vitro; Inherited; Intervention; Ions; Life; Matrix Metalloproteinases; Mechanics; Mediating; Methods; Modeling; Morphogenesis; Natural regeneration; Operative Surgical Procedures; Outcome; Phenotype; Process; Production; Protein-Lysine 6-Oxidase; Proteins; Pump; Rattus; Recruitment Activity; Relative (related person); Reporting; Resistance; Rubber; Rupture; Salts; Shapes; Signal Pathway; Signal Transduction; Site; Smooth Muscle Myocytes; Staging; Stretching; Structure; Testing; Time; Tissue Engineering; Tissues; Tropoelastin; United States; Work; Wound Healing; Xenograft procedure; base; cell assembly; cell behavior; controlled release; crosslink; cytotoxicity; density; effective therapy; extracellular; flexibility; healing; improved; in vitro Model; in vivo; innovation; nanoparticle; precursor cell; public health relevance; receptor; regenerative; repaired; response; response to injury; scaffold; tool; treatment strategy

DESCRIPTION (provided by applicant): Abdominal aortic aneurysms (AAAs), characterized by degrading aortic elastin matrix and resultant vessel weakening and rupture, causes 15,000 deaths in the United States anually, primarily amongst seniors, and those suffering from inherited matrix disorders. Regression of existing aneurysms by restoring healthy elastin architecture is difficult since adult cells poorly synthesize elastin and no tools are available to induce faithful elastin regeneration. Thus, our long-term goal is to investigate strategies to enable elastin regeneration within AAAs, so as to delay or eliminate surgical intervention. We determined that elastogenic cues based on tetramers of hyaluronan (HA), a matrix glycosaminoglycan, and TGF-2 synergestically upregulate elastin matrix synthesis and assembly by healthy adult vascular smooth muscle cells, and to a lesser extent by aneurysmal cells. The outcomes portend tremendous utility of these cues to similar elastin regeneration within AAs. However, recruitment and crosslinking of soluble elastin precursors into a stable matrix is inefficient and must be up-regulated, an insufficiency we propose to address. Our objective is thus to investigate impact of such cues (LOX, an elastin crosslinking enzyme, and Cu2+ ions), provided concurrent to or independent of elastogenic cues, on elastin synthesis, matrix assembly, and cell phenotype (e.g., elastase and MMP release) by cultured healthy and aneurysmal adult rat aortic SMCs (RASMCs). We also seek to investigate the efficacy of the optimized cues for elastin matrix regeneration in induced rat AAAs. In each of three proposed aims, we will in parallel investigate an `endogenous model' of elastin matrix regeneration, wherein RASMCs (healthy and aneurysmal) will be provided cues to both synthesize, and assemble and crosslink tropoelastin precursors. In the exogenous model, SMCs (healthy and aneurysmal) will be provided LOX and Cu2+ cues only to upregulate cellular assembly and maturation of exogenous tropoelastin. Aim 1 will investigate dose-specific benefits of exogenous, LOX to elastin synthesis, matrix assembly, and cell phenotype, within RASMC cultures. Aim 2 will evaluate dose-specific effects of copper ion delivery from copper nanoparticles (CuNP), concurrent with optimized LOX cues, to LOX activity and to elastin synthesis and matrix assembly and cell phenotype within RASMC cultures. Finally, aim 3 will test utility of optimized crosslinking cues (LOX, Cu2+) for in situ cellular assembly of elastin matrices, to stabilize induced rat aortic aneurysms (AAs) in various stages of development, when tropoelastin is (A) endogenously prompted by elastogenic cues, or (B) exogenously supplied. We expect the project outcomes to offer more effective treatment options for AAAs, based on both in situ regeneration and stabilization of elastin matrices that may be employed as a stand-alone option or in consort with existing surgical or future pharmacological approaches. Other applications of the project outcomes, specifically those pertaining to studies of healthy vascular cells, include augumenting elastin synthesis, assembly, and matrix quality within tissue engineered constructs, restoring elastin homeostasis in de-elasticized vascular allografts and xenografts, and possibly even serving as in vitro models to investigate elastogenesis during early morphogenesis, and wound healing in adult vessels. PUBLIC HEALTH RELEVANCE: Abdominal aortic aneurysms (AAAs) are potentially fatal conditions afflicting major blood vessels, which are characterized by a loss of blood vessel wall flexibility, and their ultimate structural weakening and rupture. This occurs due to breakdown and loss of rubber-like protein fibers (elastin) that normally help vessels restore their shape and form after deformation. Since cells within blood vessels cannot themselves produce new elastin, this study proposes to provide cells in culture, or within living blood vessels, a combination of biological molecules that will either induce cells to (a) synthesize new soluble elastin building blocks (precursors) and further assemble them into fiber structures, or (b) only assemble elastin precursors that are also simultaneously provided to them. The project outcomes can significantly benefit the development of new, non-surgical treatment strategies that can halt progress of or even regress existing AAAs by coaxing cells within to regenerate new elastin structures or repair and stabilize existing ones.

描述（由申请人提供）：腹主动脉瘤（AAA）的特征是降解主动脉弹性蛋白基质和导致的血管减弱和破裂，在美国造成15,000例死亡，主要是老年人，以及患有遗传性基质疾病的人。通过恢复健康的弹性蛋白结构来回归现有动脉瘤，因为成年细胞质量不佳，弹性蛋白的合成且没有任何工具可诱导忠实的弹性弹性再生。因此，我们的长期目标是调查使AAAS内弹性蛋白再生的策略，以延迟或消除手术干预。我们确定基于透明质酸（HA）的四聚体，基质糖胺聚糖和TGF-2协同上调弹性蛋白基质合成和由健康的成年血管平滑肌细胞组装的弹性提示，并通过Aneurysmal细胞降低。这些结果将这些线索的巨大效用预示到AAS内类似的弹性蛋白再生。但是，将可溶性弹性蛋白前体募集和交联成稳定的基质效率低下，必须上调，这是我们建议解决的不足。 Our objective is thus to investigate impact of such cues (LOX, an elastin crosslinking enzyme, and Cu2+ ions), provided concurrent to or independent of elastogenic cues, on elastin synthesis, matrix assembly, and cell phenotype (e.g., elastase and MMP release) by cultured healthy and aneurysmal adult rat aortic SMCs (RASMCs).我们还试图研究优化提示在诱导大鼠AAAS中弹性蛋白基质再生的功效。在提出的三个目标中的每一个中，我们将同时研究弹性蛋白基质再生的“内源性模型”，其中RASMC（健康和动脉瘤）将得到合成，并组装和交叉链接链球菌素前体的提示。在外源模型中，将提供SMC（健康和动脉瘤），仅提供LOX和CU2+提示，以上调外源性Tropoelastin的细胞组件和成熟。 AIM 1将研究RASMC培养物中外源性，LOX对弹性蛋白合成，基质组装和细胞表型的特异性益处。 AIM 2将评估铜离子从铜纳米颗粒（CUNP）递送的剂量特异性作用，并同时发生优化的LOX提示，转化为LOX活性，以及​​弹性蛋白合成，矩阵组装和RASMC培养物中的细胞表型。最后，AIM 3将测试优化的交联提示（LOX，CU2+）的实用性，以便在弹性蛋白基质的原位细胞组装中，以稳定诱导的大鼠主动脉瘤（AAS）在各个发育阶段，当tropoelastin是（a）由（a）源自弹性的提示，或（a）由弹性基因提示，或（a）由弹性基因提示。我们预计，基于原位再生和稳定弹性蛋白矩阵，这些项目的结果将为AAA提供更有效的治疗选择，这些弹性矩阵可以用作独立选择，或者采用现有的外科手术或将来的药理学方法。项目结果的其他应用，特别是与健康血管细胞研究有关的结果，包括在组织工程结构内进行弹性蛋白合成，组装和基质质量，恢复去弹性的血管稳态中的弹性蛋白稳态，以及在较早的较早的情况下，还可以调查伊利诺伊特的弹性同种异体移植物，甚至可能是培养伊斯特罗的疾病。公共卫生相关性：腹部主动脉瘤（AAAS）是潜在的致命疾病，遭受了主要血管，其特征是血管壁的柔韧性丧失，以及它们的最终结构性弱化和破裂。这是由于分解和橡胶样蛋白纤维（弹性蛋白）的损失，通常帮助血管在变形后恢复其形状和形成。由于血管中的细胞本身不能产生新的弹性蛋白，因此本研究提议在培养物或活血管中提供细胞，这是生物分子的组合，它们将诱导细胞诱导细胞（a）合成新的可溶性弹性弹性蛋白构建块（前体）（前体），并进一步将它们组装到纤维结构中，或者（b）仅提供弹性蛋白前蛋白的前列体。该项目的结果可以显着有益于新的非手术治疗策略的制定，这些策略可以通过哄骗内部的细胞来再生新的弹性蛋白结构或修复并稳定现有的弹性，从而阻止现有的AAA的进展甚至会回归现有的AAA。