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

项目摘要 不愈合溃疡是糖尿病的常见并发症，导致生活质量下降， 截肢，增加死亡风险，以及高昂的医疗费用。不幸的是，目前的治疗方法仍然 过时和不足。在糖尿病中，真皮组织中的神经病变和微血管变化导致 失调的分子线索，导致慢性炎症和减少的血管生成，防止伤口 治愈考虑到脉管系统在供氧中的重要性，不良的血管生成是特别关键的， 营养素和系统信号分子。血管生成的损害部分是由异常表达驱动的 编码信使RNA（mRNA）和非编码微RNA（miRNA）在不同时间尺度上的变化。因此， 改变糖尿病溃疡病程的一种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，递送挑战使得核酸RNA靶向疗法在非愈合状态下不起作用。 酸疗法在临床上不可行。为了应对这些交付挑战，哈蒙德实验室开发了 通过逐层（LbL）技术证明了核酸的自组装静电沉积， 其利用交替电荷的聚电解质的迭代吸附，以在 具有可调释放动力学的伤口绷带。我建议开发和研究时间控制的 释放策略，以局部递送RNA靶向治疗，促进血管生成和愈合， 糖尿病溃疡在目标1中，我将制定阶段性释放RNA靶向绷带，以促进伤口愈合 因为对多个目标的分阶段释放治疗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，用于靶向治疗以刺激血管生成 在炎症和增生性伤口愈合阶段，将在体外和体内测试其功效。 小鼠体内糖尿病溃疡模型。在目标2中，我将确定促血管生成抗miR的潜在协同作用 （miRNA抑制剂），因为用抗miR抑制基因表达使得能够沿着调节许多基因。 定义了组织特异性信号通路以增强血管生成。由于也不知道交货时间 这些抗miR组合可能会影响疗效，我们将利用控释LbL绷带， 调查这个。通过这项研究，我将推进核酸与生物材料系统的交付， 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 在愈合过程中的定时输送至关重要的再生，如静脉溃疡、肠系膜溃疡、 局部缺血和心肌梗塞。

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 Unfortunately, current treatments remain outdated and inadequate.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.