Vascular Endothelial Dysfunction in Preeclampsia

先兆子痫的血管内皮功能障碍

• 批准号: 8136433
• 负责人: IAN M. BIRD
• 金额: $ 22.28万
• 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 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8136433
• 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. PUBLIC HEALTH RELEVANCE: Preeclampsia (PE) is a potentially devastating disease of pregnancy that can threaten the life of both mother and the unborn child. It affects as many as 20% of the population. Even mild PE is dangerous and can cause preterm (early) birth and this then gives the baby an increased risk to develop several diseases such as blood pressure, diabetes and obesity as an adult. We have shown in mothers with PE that the cells lining the umbilical cord blood vessels have reduced function and so may be a readily available and useful tool to screen dugs for therapy. The aim of this project is to show our suspicions are correct. If we are right we may be able to come up with a drug-screening plan to develop future treatment. Having more complete knowledge of how cell function is regulated in umbilical vein from both normal and PE pregnancy will certainly take us one step closer to overcoming this potentially devastating disease that effects so many in this country and beyond.

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尚不清楚。当前有关相关高血压的疗法仍主要针对血管平滑肌的放松，但这不足以完全控制该疾病。基于新颖的治疗干预方法，需要采取新的策略。只有在明确识别的新治疗靶标和在内皮功能级别进行筛选的方法只有可能。这样的大规模筛选方法需要大量的高纯度，来自PE受试者的细胞表征良好。然后，在抗药性水平的第二轮测试中，可以在逼真的范围内进行逼真的量表，然后再进行人类受试者的药物细化和试验。问题仍然是，从正常和PE妊娠的人子宫动脉组织不足以进行初始筛查，而较小的血管活检样品不足以产生足够纯度或数量的细胞来筛查药物面板筛查。我们的初步数据表明，尽管在正常怀孕的子宫动脉中确实最大化了血管内皮功能，但在其他血管层中也观察到这些相同的机制，并最大化，包括胎儿静脉（例如脐静脉）。此外，这些信号事件和在正常绳索内皮中没有产生的没有产生的PE受试者似乎也失败了。因此，为了确定在这样的药物筛查中是否可以将PE受试者的HUVEC用作合适的细胞，我们提出：Spaim 1：假设：观察到的持续[Ca2+] I在正常情况下对Intact UV Endo中的ATP响应爆发。怀孕是由于TRPC3和CX43激活，以及与针对TRPC3或CX43抑制的策略相关的Ca2+爆发的损失，可复制较低/钝的Ca2+，并且在PE受试者中看不到反应。 SPAIM 2（向目标3）：假设：与正常受试者相比，PE衍生的紫外线内部或PE衍生的HUVEC（通道3）中PE相关的功能障碍并不是由于P2Y2受体的相对水平G，G蛋白质，PLC，IP3受体，TRPC通道或连接素。 SPAIM 3：假设：在短期培养中，由于CX43促进IP3敏感的TRPC3激活而导致的持续的[Ca2+] I对HUVEC的ATP持续反应，并且缺乏从PE受试者获得的细胞。此外，抑制IP3介导的TRPC3激活或正常受试者中HUVEC中的CX43介导的细胞 - 细胞通信会复制PE受试者HUVEC中的功能丧失，但不会进一步抑制PE受试者HUVEC的功能。鉴于成功，PE受试者的脐带的现成可用性和大量的紫外线内部细胞意味着可以进行未来的大规模药物测试，即使额外关注的是针对性/不利的公众亚种群。这可以允许对新药或现有药物的新应用进行更有选择的干预，从而减轻先兆子痫的破坏性影响。 公共卫生相关性：先兆子痫（PE）是一种可能造成毁灭性的怀孕疾病，可能威胁母亲和未出生的孩子的生命。它影响了多达20％的人口。即使是轻度的PE也很危险，可能会导致早产（早产），然后这使婴儿增加了多种疾病的风险，例如血压，糖尿病和成年人的肥胖症。我们已经在患有PE的母亲中表明，脐带血血管内壁的细胞降低了功能，因此可能是一种可用且有用的工具，可用于筛选治疗的挖掘。该项目的目的是表明我们的怀疑是正确的。如果我们是正确的，我们可能会提出制定筛查计划来制定未来的治疗。对从正常和PE妊娠中的脐带静脉中如何调节细胞功能的情况更加完整，肯定会使我们更近一步地克服了这种潜在的毁灭性疾病，这种疾病在这个国家及以后会影响很多。