Defining the complex biology of the miR-130/301 family in pulmonary hypertension

定义 miR-130/301 家族在肺动脉高压中的复杂生物学

• 批准号: 8914034
• 负责人: Stephen Y Chan
• 金额: $ 7.22万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2015-09-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8914034
• 关键词: Atomic Force Microscopy; Binding; Bioinformatics; Biology; Blood Vessels; Cell Proliferation; Collagen; Complex; Data; Deposition; Development; Disease; Endothelin-1; Family; Family member; Foundations; Future; Gene Targeting; Generations; Genes; Genetic; Health; Human; Individual; Life; Link; Lung; Mammals; Matrix Metalloproteinases; Measurement; Measures; Methods; MicroRNAs; Microscopy; Modeling; Molecular; Mus; Network-based; Oligonucleotides; PPAR gamma; Pathway interactions; Physiological; Process; Pulmonary Hypertension; Pulmonary vessels; RNA; RNA Interference; Regulation; Signal Pathway; Signal Transduction; Smooth Muscle; System; TIMP2 gene; Techniques; Time; Transgenes; Validation; Vascular Diseases; Vasomotor; Viral; base; combinatorial; constriction; disorder control; hemodynamics; in vivo; intravital microscopy; new therapeutic target; pressure; synergism; therapeutic miRNA; vasoconstriction

DESCRIPTION (provided by applicant): Pulmonary hypertension (PH) is a deadly vascular disease with enigmatic molecular origins. While individual microRNAs (miRNAs) can control PH modestly, related miRNAs may conspire to regulate multiple target gene "networks" for a more robust influence on disease. Yet, the network biology of miRNAs has been poorly explored in vivo. We hypothesize that the miR-130/301 family is a pathogenic lynchpin controlling numerous target pathways (beyond pro-proliferative signaling) for broad control over PH. We will define mechanisms connecting the miR-130/301 family to pulmonary vascular dysfunction in vivo - via possible targeting of PPARγ and TIMP2 to alter vasomotor tone and vascular stiffness. We will also delineate individual actions of miRNA family members and their coordinated effects in vivo. Thus, the redundant/ synergistic effects thought to be active in miRNA biology will be examined for the first time in PH in vivo. Specific Aims: Aim 1) Determine the importance of PPARγ and endothelin-1 for control of pulmonary vasomotor tone by miR-130/301. To prove whether miR-130/301 causes vasoconstriction via regulating PPARγ and endothelin-1 (ET-1), miR-130/301, PPARγ, and ET-1 will be modulated in mice using systems of RNAi and adeno-associated viral (AAV) transgene delivery to the pulmonary vessels, along with intravital pulmonary vascular microscopy and pressure-flow curve analyses to measure vasomotor tone directly. Aim 2) Determine the importance of PPARγ and TIMP2 for control of matrix deposition and pulmonary vascular stiffness by miR-130/301. We postulate that miR-130/301 further controls PH manifestation via repressing both PPARγ and TIMP2, thus increasing matrix deposition and vascular stiffness. We will modulate miR-130/301, PPARγ, and TIMP2 in mice, followed by biophysical (atomic force microscopy) and molecular assessment of pulmonary vascular stiffness. Results could provide the first evidence in vivo of miRNA-specific regulation of pulmonary vascular stiffness and combinatorial control of PH via multiple related miRNA targets. Aim 3) Define the coordinated actions of the miR-130/301 family members on overall PH manifestation. We postulate that the miR-130/301 family employs both synergism and redundancy to promote PH. An optimized system of miRNA delivery to the pulmonary vasculature in vivo will be used systematically to interrogate the individual versus integrated contributions of each miR-130/301 family member to PH manifestation in mice. This study will be the first to define the specific network-based parameters of miRNA biology in control of PH in vivo and could be a necessary guide in developing miRNA-based therapies of PH. Significance: Our proposal incorporates a rigorous expertise in miRNA biology with new technological advancements in the molecular, biophysical, and physiological study of PH in vivo. Thus, we aim to establish miR-130/301 as a multi-faceted regulator of PH -- offering new therapeutic targets and perhaps accelerating discovery in other diseases that rely upon complex miRNA networks for control of resultant pathophenotypes.

描述（由适用提供）：肺动脉高压（pH）是一种致命的血管疾病，具有神秘的分子起源。虽然单个microRNA（miRNA）可以适度控制pH，但相关的miRNA可能会合并调节多个靶基因“网络”，从而对疾病产生更大的影响。然而，miRNA的网络生物学在体内探索了很差。我们假设miR-130/301家族是一种致病性的林奇宾，它控制着许多目标途径（超出促进信号传导），以对pH的广泛控制。我们将通过可能靶向PPARγ和TIMP2来改变血管舒缩张力和血管僵硬，将MiR -130/301家族与体内肺血管功能障碍的机制定义为肺血管功能障碍。我们还将描述miRNA家族成员的个人行为及其在体内的协调效果。这，将首次在体内pH值检查miRNA生物学中的冗余/协同作用。具体目的：目标1）确定miR-130/301通过miR-130/301来控制PPARγ和内皮素-1对肺血管舒张张张的重要性。证明miR-130/301是否通过调节PPARγ和内皮蛋白-1（ET-1），miR-130/301，PPARγ和ET-1是否引起血管收缩血管舒张态直接。目标2）确定miR-130/301的PPARγ和TIMP2对控制基质沉积和肺血管刚度的重要性。我们假设miR-130/301通过反映PPARγ和TIMP2进一步控制pH表现，从而增加了基质沉积和血管刚度。我们将在小鼠中调节miR-130/301，PPARγ和TIMP2，然后进行生物物理（原子力显微镜）和肺血管刚度的分子评估。结果可以通过多个相关的miRNA靶标的miRNA特异性调节肺血管刚度和组合控制的miRNA特异性调节。目标3）定义miR-130/301家庭成员在总体pH表现方面的协调行动。我们假设MiR-130/301家庭员工既有协同作用和冗余，又可以促进PH。在体内将miRNA递送到肺脉管系统的优化系统将系统地用于询问每个miR-130/301家族成员对小鼠中pH表现的单个和综合贡献。这项研究将是第一个定义miRNA生物学的特定基于网络的参数，以控制体内pH值，并且可能是开发基于miRNA的pH疗法的必要指南。意义：我们的建议将miRNA生物学的严格专业知识与体内pH的分子，生物物理和物理研究方面的新技术进步结合在一起。这是我们的目标是将miR-130/301建立为pH的多方面调节剂 - 提供新的治疗靶标，并可能加速其他疾病中的发现，这些疾病依靠复杂的miRNA网络来控制产生的病理表型。