Repairing the Kidney Endothelium via Targeted Extracellular Matrix Modifiers

项目总结 肾微血管是炎症性疾病和缺氧性损伤的汇聚点。 内皮调节失调和潜在的细胞外基质(ECM)恶化。加在一起，这些 这些变化会导致进行性肾功能障碍，最终导致衰竭。微血管的调节作用 一般来说--尤其是肾脏的微血管--超出了它的载血能力， 对全身动态平衡有全球影响。尽管有肾血管动物模型的发展 功能障碍，这是科学研究、治疗工具的翻译和 从动物到人类的知识受到转基因疾病模型之间不一致联系的限制 和人类血管生理学。对肾血管微环境的新的科学认识 细胞外基质的组成及其内皮细胞的相互依赖为体外发育提供了信息 肾微血管模型。然而，生物工程系统往往过度简化复杂的， 肾内皮细胞生物学的相互依赖性质以及与肾内周细胞和间质的必要的相互作用 微环境。尽管在光刻和添加剂制造方面取得了新的进展，但微小的长度 通常在体内微血管系统中发现的鳞片不能复制，因此不能充分 概述体外肾脏微环境。为了解决这一缺陷，我们的多学科团队开发了 一种模仿正常肾脏的鳞片、ECM组成和流体力学的生物复制型肾脏微血管 肾脏。科学调查的基础是这个血管化的脚手架系统，由 我们发表的数据显示了动脉和静脉循环的专利和可灌注性(Caralt等人，Am J 移植，2015)，具有严格的流体动力学控制(尤扎斯基等人，组织工程C部分方法，2015) 这两者共同产生了一个生物复制的“试验台”。该平台提供了操纵ECM的独特机会 利用新技术解锁细胞功能的微环境。为了能够进行这样的调查，我们 开发了可定向ECM修饰剂(TEMS)，这是一种基于我们初步研究的生物材料递送系统 和发布的数据(酱等人生物大分子，2016)展示了区别靶向和 将生物活性物质运送到特定的ECM子组件。我们的假设是内皮修复导致 血管修复可以通过将生物活性物质输送到具有特异性影响的基质中来控制 ECM界面的内皮细胞。我们的调查得到了数据的支持，数据显示增长了7倍 与在溶液中自由传递的可溶性因子相比，TEM访问的ECM支架内的因子领先 在没有血管内皮细胞的情况下，体外血管内皮细胞维持28天。这 生物工程、纳米技术、 多肽化学和肾脏学：用一组TEM定制体外肾血管系统以开发测试 研究修复肾血管损伤和逆转肾脏疾病的疾病和治疗方法的平台。

PROJECT SUMMARY The renal microvasculature is the convergence point for inflammatory disorders and hypoxic injury that cause endothelial dysregulation and deterioration of the underlying extracellular matrix (ECM). Together, these changes lead to progressive kidney dysfunction and ultimately failure. The regulatory role of the microvasculature in general—and in particular the microvasculature of the kidney—extends beyond its blood carrying capacity, with global implications to total-body homeostasis. Despite development of animal models of renal vascular dysfunction, which are important components of scientific research, translation of therapeutic tools and knowledge from animals to humans is limited by inconsistent linkages between transgenic models of disease and human vascular physiology. New scientific understanding of the renal vasculature microenvironment, its ECM composition, and the interdependency of endothelial cells within it provide information to develop ex vivo models of renal microvasculature. However, bioengineered systems oftentimes oversimplify the complex, interdependent nature of renal endothelial biology and the necessary cross-talk with pericytes and stroma within the microenvironment. Despite new advances in photolithography and additive manufacturing, the tiny length scales typically found within the in vivo microvasculature cannot be replicated and thus fail to adequately recapitulate the renal microenvironment ex vivo. To address this deficiency, our multidisciplinary team developed a bio-replicative renal microvasculature that mimics the scale, ECM make-up and fluid mechanics of the normal kidney. The foundation for the scientific investigation is this vascularized scaffolding system that is supported by our published data demonstrating patent and perfusable arterial and venous circulation (Caralt et al., Am J Transplant, 2015) with strict control of hydrodynamics (Uzarski, et al., Tissue Eng Part C Methods, 2015) that together result in a bio-replicative ‘test rig’. This platform provides unique opportunities to manipulate the ECM microenvironment with new technologies that unlock cellular function. To enable such an investigation, we developed Targetable ECM Modifiers (TEMs), a new biomaterial delivery system based upon our preliminary and published data (Jiang, et al. Biomacromolecules, 2016) demonstrating ability to discriminately target and shuttle bioactive agents to specific ECM sub-components. Our hypothesis is that endothelial repair leading to vascular restoration can be controlled by delivering bioactive materials to the matrix with specificity to influence endothelial cells at ECM interfaces. Our investigation is supported by data showing a 7-fold enrichment of growth factors within ECM scaffolds accessed by TEMs, compared to soluble factors delivered free in solution, leading to maintenance of an ex vivo vascular endothelium for 28 days where none developed in its absence. This investigation is further enabled by a multidisciplinary team of collaborators in bioengineering, nanotechnology, peptide chemistry and nephrology to tailor the ex vivo renal vasculature with a panel of TEMs to develop testing platforms to study disease and therapies to repair renal vascular injury and reverse kidney disease.