MicroRNA 93 in Peripheral Arterial Disease

MicroRNA 93 在外周动脉疾病中的作用

基本信息

批准号：
8896862
负责人：
BRIAN H ANNEX
金额：
$ 38.91万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8896862
关键词：
Affect American Ankle Apoptosis Atherosclerosis Back Binding Blood Pressure Blood Vessels Blood flow Cell Count Cell Cycle Cells Cholesterol Clinical Complication DNA Data Development Distal E2F1 gene Educational process of instructing Endothelial Cells Endothelium Exercise Gene Expression Genes Goals Growth Health Homing Human Hypoxia In Vitro Inbred BALB C Mice Inbred Strains Mice Investigation Ischemia Knockout Mice Laboratories Leg Limb structure Link Lower Extremity Measures Medical Messenger RNA Methods MicroRNAs Mus Muscle Muscle Cells Necrosis Nucleotides Pathway interactions Patients Perfusion Peripheral arterial disease Plasma Play Pre-Clinical Model Process Proteins RNA Recovery Regulator Genes Role Sampling Serum Severities Site Skeletal Muscle Starvation Stem cells Symptoms TP53 gene Testing Therapeutic Therapeutic Agents Time Tissues Training Translating Translations Umbilical vein Up-Regulation angiogenesis base cdc Genes cell type design differential expression disorder control gain of function gene therapy improved in vivo indexing loss of function mRNA Expression novel therapeutic intervention novel therapeutics response response to injury skeletal

项目摘要

DESCRIPTION (provided by applicant): Peripheral arterial disease (PAD) is a major complication of systemic atherosclerosis which affects approximately 12 million Americans. The lower extremity is the most common site for PAD where current medical treatments target systemic atherosclerosis but are not able to improve perfusion to the ischemic limb and directly treat the problem in PAD. There is a pressing clinical need for therapeutic approaches for PAD. Though only fully recognized as a true biologic entity less than 15 years ago, micro-RNAs (miRs) which are 15 - 23 nucleotide long RNA's, are now appreciated to be key regulators of gene expression; especially in the adaptive response to injury. The best characterized method by which miRs regulate gene expression is by their ability to bind to the 3' un-translated region of a target mRNA and thereby reducing mRNA expression or protein translation. However, a single miR can regulate entire biologic pathways by affecting several functionally related genes thus making miRs attractive therapeutic agents. We found that miR-93 appears to play a critical role in experimental PAD. Following HLI, C57BL/6 (B6) mice have excellent perfusion recovery and little tissue loss. In contrast, in Balb/C mice perfusion recovery is limited and necrosis is common. Utilizing both experimental and computational approaches we identified mir- 93 as the miR that was MOST differentially expressed based on the combination of strain and ischemia. miR- 93 was significantly increased following HLI (p<0.0001 and >4-fold increase) in B6 mice not in Bab/C. Local delivery of a pre-miR93 led to greater perfusion recovery in Balb/C mice and systemic delivery of a miR-93 antagomer impaired perfusion recovery in C57/Bl6 mice. In-vitro, antagomers to miR-93 increased the extent of hypoxia induced apoptosis while treatment with pre-miR-93 reduced the extent of hypoxia-induced apoptosis in both cell types. Antagomers to miR-93 reduced, and administration of pre-miR 93 increased, proliferation. We consistently found that the cycle genes p53, E2F1, and p21 were altered. Collectively, central hypothesis that miR-93, can serve as a potent modulator of post-natal angiogenesis in PAD though its ability to regulate cycle genes alone, or with other potential targets. Specific Aim 1): Define the role that miR-93 plays in-vivo in regulating the extent of blood flow recovery post-HLI using gain of function and loss of function approaches by the examination of mice at appropriate pre-selected time points. Specific Aim 2): Utilize in-vitro studies to determine the extent to which th gene expression changes induced by miR-93 can be directly linked to miR-93/mRNA interactions and establish the extent to which the gene expression changes induced by miR-93 are cell and condition specific. Specific Aim 3): Measure miR-93 in an established bank of human plasma and skeletal human samples from patients with varying severity of PAD as well as a group of non-PAD controls.

描述（由申请人提供）：外周动脉疾病 (PAD) 是全身动脉粥样硬化的主要并发症，影响大约 1200 万美国人。下肢是 PAD 最常见的部位，目前的医学治疗针对的是全身动脉粥样硬化，但无法改善缺血肢体的灌注并直接治疗 PAD 问题。临床迫切需要 PAD 的治疗方法。虽然不到 15 年前才被充分认识为真正的生物实体，但 micro-RNA (miR)（15 - 23 个核苷酸长的 RNA）现在被认为是基因表达的关键调节因子。尤其是对伤害的适应性反应。 miR 调节基因表达的最佳表征方法是通过它们与靶 mRNA 的 3' 非翻译区结合的能力，从而减少 mRNA 表达或蛋白质翻译。然而，单个 miR 可以通过影响几个功能相关的基因来调节整个生物途径，从而使 miR 成为有吸引力的治疗剂。我们发现 miR-93 似乎在实验性 PAD 中发挥着关键作用。 HLI 后，C57BL/6 (B6) 小鼠具有出色的灌注恢复和很少的组织损失。相比之下，Balb/C 小鼠的灌注恢复有限且坏死很常见。利用实验和计算方法，我们将 mir-93 确定为基于应变和缺血的组合表达差异最大的 miR。 HLI 后，B6 小鼠中的 miR-93 显着增加（p<0.0001 且增加 >4 倍），而 Bab/C 中则不然。局部递送 pre-miR93 导致 Balb/C 小鼠更大的灌注恢复，而全身递送 miR-93 拮抗剂则损害 C57/Bl6 小鼠的灌注恢复。在体外，miR-93 的拮抗剂增加了缺氧诱导的细胞凋亡的程度，而前 miR-93 的治疗则降低了两种细胞类型缺氧诱导的细胞凋亡的程度。 miR-93 的拮抗剂减少，并且前 miR 93 的施用增加增殖。我们一致发现循环基因 p53、E2F1 和 p21 发生了改变。总的来说，核心假设是 miR-93 通过单独或与其他潜在靶标一起调节周期基因的能力，可以作为 PAD 出生后血管生成的有效调节剂。具体目标 1)：定义通过在适当的预先选择的时间点检查小鼠，使用功能获得和功能丧失方法来研究 miR-93 在体内调节 HLI 后血流恢复程度的作用。具体目标 2)：利用体外研究确定 miR-93 诱导的基因表达变化与 miR-93/mRNA 相互作用直接相关的程度，并确定 miR-93 诱导的基因表达变化的程度。 93 是特定于细胞和条件的。具体目标 3)：测量已建立的人类血浆和骨骼样本库中的 miR-93，这些样本来自不同严重程度的 PAD 患者以及一组非 PAD 对照。