Vascular Endothelial Dysfunction in Preeclampsia

先兆子痫的血管内皮功能障碍

• 批准号: 8291197
• 负责人: IAN M. BIRD
• 金额: $ 18.56万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8291197
• 关键词: Adult; Affect; Arteries; Biopsy Specimen; Birth; Blood Pressure; Blood Vessels; Cell Communication; Cell Culture Techniques; Cell Line; Cell physiology; Cells; Chemosensitization; Connexin 43; Connexins; Country; Coupling; Data; Diabetes Mellitus; Disadvantaged; Disease; Dyes; Endothelial Cells; Endothelium; Event; Failure; Functional Imaging; Functional disorder; Future; G alpha q Protein; GTP-Binding Proteins; Human; Hypertension; Image; Intervention; Knowledge; Life; Mediating; Methodology; Modeling; Molecular; Mothers; Myography; Normal tissue morphology; Obesity; One-Step dentin bonding system; P2Y2 receptor; Pharmaceutical Preparations; Placenta; Population; Pre-Eclampsia; Preclinical Drug Evaluation; Pregnancy; Preparation; Production; Proteins; Relative (related person); Relaxation; Resistance; Risk; Screening procedure; Side; Signal Pathway; Signal Transduction; Testing; Therapeutic Intervention; Time; Umbilical Cord Blood; Umbilical vein; Vascular Smooth Muscle; Work; base; disorder control; drug testing; fetal; human subject; inhibitor/antagonist; loss of function; new therapeutic target; novel strategies; receptor; response; success; tool; unborn child; vascular bed; vascular endothelial dysfunction

DESCRIPTION (provided by applicant): While endothelial dysfunction is known to be a part of preeclampsia (PE), the molecular basis for endothelial dysfunction remains unclear. Current therapies for the associated hypertension are still largely targeted to relaxation of vascular smooth muscle and yet this is not sufficient to fully control the disorder. New strategies are needed based on novel approaches to therapeutic intervention. This will only be possible given a clearly identified new therapeutic target and a means to screen it at the level of endothelial function. Such large scale screening approaches require large numbers of high purity, well characterized cells from PE subjects. Second round testing in resistance level vessels can then be undertaken on a realistic scale before moving towards drug refinement and trials in human subjects. The problem remains that human uterine artery tissue from normal and PE pregnancy are not sufficiently available to undertake the initial screen, and smaller vessel biopsy samples are insufficient to yield cells of high enough purity or quantity for drug panel screening. Our preliminary data suggests that while vascular endothelial function is indeed maximized in the uterine artery in normal pregnancy, these same mechanisms are also observed and maximized in other vascular beds including fetoplacental vessels such as the umbilical vein. Further, these signaling events and associated NO production in the endothelium of normal cords also appear to have failed in PE subjects. Thus, in order to establish if it is possible to use HUVEC from PE subjects as suitable cells in such a drug screen we propose: SpAim 1: Hypothesis: The observed sustained [Ca2+]i bursts in responses to ATP in intact UV Endo from normal pregnancy are due to TRPC3 and CX43 activation, and losses of Ca2+ burst associated with strategies aimed at TRPC3 or CX43 inhibition replicate the lower/blunted Ca2+ and NO responses seen in PE subjects. SpAim 2 (Transitional to Aim 3): Hypothesis: PE associated dysfunction in PE derived UV Endo or PE derived HUVEC (passage 3) compared to that from normal subjects is not due to simple changes in the relative levels of expression of P2Y2 receptors, G proteins, PLC, IP3 receptors, TRPC channels, or connexins. SpAim 3: Hypothesis: In short term culture, the sustained [Ca2+]i burst responses to ATP in HUVEC from normal pregnancy due to CX43 potentiation of IP3 sensitive TRPC3 activation are retained, and are lacking in cells derived from PE subjects. Further, inhibition of either IP3 mediated TRPC3 activation or CX43 mediated cell-cell communication in HUVEC from normal subjects replicates the loss of function seen in HUVEC from PE subjects, but does not further inhibit the function of HUVEC from PE subjects. Given success, the ready availability of cords and large yields of UV Endo cells from PE subjects means that future large scale drug testing could be performed, even with additional focus on targeted/disadvantaged subpopulations of the public. This could allow more selective intervention with new drugs or new applications of existing drugs that impact on this signaling pathway, and so relieve the devastating effects of preeclampsia.

描述（由申请人提供）：虽然已知内皮功能障碍是先兆子痫（PE）的一部分，但内皮功能障碍的分子基础仍不清楚。目前对相关高血压的治疗仍然主要针对血管平滑肌的松弛，但这不足以完全控制疾病。需要基于新的治疗干预方法的新策略。只有在明确确定了新的治疗靶点并在内皮功能水平上对其进行筛选的情况下，这才有可能。这种大规模筛选方法需要来自PE受试者的大量高纯度、充分表征的细胞。然后可以在现实的规模上进行耐药水平血管中的第二轮测试，然后再进行药物精制和人类受试者试验。问题仍然是，来自正常和PE妊娠的人子宫动脉组织不足以进行初始筛选，并且较小的血管活检样品不足以产生用于药物组筛选的足够高纯度或数量的细胞。我们的初步数据表明，虽然血管内皮功能在正常妊娠的子宫动脉中确实最大化，但在其他血管床（包括胎儿胎盘血管，如脐静脉）中也观察到并最大化了这些相同的机制。此外，这些信号事件和相关的NO生产在正常的脐带内皮细胞也似乎已经失败的PE受试者。因此，为了确定是否可以使用来自PE受试者的HUVEC作为这种药物筛选中的合适细胞，我们提出：在正常妊娠的完整UV Endo中观察到的响应ATP的持续[Ca 2 +]i爆发是由于TRPC 3和CX 43激活，与TRPC 3或CX43抑制策略相关的Ca 2+爆发损失复制了PE受试者中观察到的较低/钝化的Ca 2+和NO应答。SpAim 2（过渡到Aim 3）：假设：与正常受试者相比，PE衍生的UV Endo或PE衍生的HUVEC（第3代）中的PE相关功能障碍不是由于P2 Y2受体、G蛋白、PLC、IP 3受体、TRPC通道或连接蛋白的相对表达水平的简单变化。SpAim 3：假设：在短期培养中，来自正常妊娠的HUVEC中由于IP 3敏感性TRPC 3活化的CX 43增强而对ATP的持续[Ca 2 +]i爆发响应被保留，并且在来自PE受试者的细胞中缺乏。此外，来自正常受试者的HUVEC中IP 3介导的TRPC 3活化或CX 43介导的细胞-细胞通讯的抑制复制了来自PE受试者的HUVEC中所见的功能丧失，但不进一步抑制来自PE受试者的HUVEC的功能。鉴于成功，来自PE受试者的脐带和大量UV Endo细胞的现成可用性意味着未来可以进行大规模药物测试，即使额外关注公众的目标/弱势亚群。这可以允许使用影响这一信号通路的新药或现有药物的新应用进行更有选择性的干预，从而减轻先兆子痫的破坏性影响。