Controlled release of RNA-targeting therapy to promote healing of diabetic ulcers

项目摘要 无法愈合的溃疡是糖尿病的常见并发症，导致生活质量下降，发病率上升。 截肢的风险增加，以及高昂的医疗费用。不幸的是，目前的治疗方法仍然 既过时又不够用。在糖尿病患者中，皮肤组织的神经病变和微血管变化导致 调节失调的分子信号，导致慢性炎症和防止伤口的血管生成减少 治愈。考虑到血管系统在供氧方面的重要性，血管生成不良尤其关键， 营养物质和系统信号分子。血管生成障碍在一定程度上是由异常表达驱动的 编码信使RNA(MRNAs)和非编码microRNAs(MiRNAs)在不同的时间尺度上。因此，一个 改变糖尿病溃疡病程的有希望的方法是直接靶向上调的RNA的表达 在使用核酸RNA靶向疗法的非愈合状态下；然而，递送挑战使核 酸性疗法在临床上是不可行的。为了应对这些交付挑战，哈蒙德实验室开发了 展示了通过逐层(LBL)技术的核酸的自组装静电沉积， 它利用交变电荷的聚电解质的迭代吸附，在 释放动力学可调的创面绷带。我建议开发和研究时间控制的 释放策略，局部提供促进血管生成和愈合的RNA靶向治疗 糖尿病溃疡。在目标1中，我将研制阶段性释放RNA靶向绷带，以促进伤口愈合 由于针对多个目标的治疗的阶段性释放将允许绷带处理伤口的不同阶段 治愈。将开发一种概念验证绷带来洗脱RNA靶向疗法以刺激血管生成 在炎症和增生性伤口愈合阶段，它将在体外和在 小鼠体内糖尿病溃疡模型的建立。在目标2中，我将确定促血管生成的抗miRs的潜在协同作用。 (miRNA抑制物)，因为用抗miRs抑制基因表达使许多基因能够在 明确了促进血管生成的组织特异性信号通路。因为它也不知道如何确定交付时间 在这些可能影响疗效的抗miR组合中，我们将利用控释LBL绷带来 调查这件事。通过这项研究，我将利用生物材料系统和 靶向异常表达的编码和非编码RNA以促进糖尿病伤口的愈合。这 这项工作将为将该平台方法扩展到其他受损组织疾病奠定基础 再生对愈合过程的时机至关重要，如静脉溃疡、肠系膜 缺血和心肌梗死。

Project Summary Non-healing ulcers are a common complication of diabetes, resulting in decreased quality of life, elevated rates of amputation, increased risk of mortality, and high healthcare costs. Unfortunately, current treatments remain outdated and inadequate. In diabetes, neuropathy and microvascular changes in dermal tissue lead to dysregulated molecular cues, resulting in chronic inflammation and reduced angiogenesis that prevent wound healing. Poor angiogenesis is particularly critical given the importance of vasculature in supplying oxygen, nutrients, and systemic signaling molecules. Impairment of angiogenesis is in part driven by aberrant expression of coding messenger RNAs (mRNAs) and non-coding microRNAs (miRNAs) at various time scales. Thus, one promising approach to alter the course of diabetic ulcers is to directly target the expression of upregulated RNAs in the non-healing state using nucleic acid RNA-targeting therapies; however, delivery challenges render nucleic acid therapies clinically unfeasible. To address these delivery challenges, the Hammond Lab has developed and demonstrated self-assembled electrostatic deposition of nucleic acids through the layer by layer (LbL) technique, which leverages iterative adsorption of polyelectrolytes of alternating charge, to create conformal coatings on wound bandages with tunable release kinetics. I propose to develop and investigate temporally controlled release strategies to locally deliver RNA-targeting therapies that promote angiogenesis and healing of diabetic ulcers. In Aim 1, I will formulate staged release RNA-targeting bandages to promote wound healing since staged release of therapy for multiple targets will allow the bandages to address different phases of wound healing. A proof-of-concept bandage will be developed to elute RNA-targeting therapy to stimulate angiogenesis in both the inflammatory and proliferative wound healing phases, and it will be tested for efficacy in vitro and in a murine in vivo diabetic ulcer model. In Aim 2, I will identify potential synergies of pro-angiogenic anti-miRs (miRNA inhibitors), as inhibition of gene expression with anti-miRs enables regulation of many genes along defined tissue-specific signaling pathways to enhance angiogenesis. Since it is also unknown how delivery timing of these anti-miR combinations may impact efficacy, we will leverage controlled-release LbL bandages to investigate this. Through this research, I will advance the delivery of nucleic acids with biomaterial systems and the targeting of aberrantly expressed coding and non-coding RNAs to promote healing of diabetic wounds. This work will lay the groundwork for expansion of this platform approach to other diseases of impaired tissue regeneration where timing the delivery to the healing process is critical, such as venous ulcers, mesenteric ischemia, and myocardial infarction.