Functional mechanism underlying Dolutegravir (DTG)-associated developmental CNS abnormalities: Inhibition of matrix metalloproteinases activities

项目摘要 背景：多洛替格雷(DTG)是一种整合酶链转移抑制剂(INSTI)，目前推荐同时使用， 治疗人类免疫缺陷的一线疗法和部分转换或挽救方案 感染1型病毒(HIV-1)的患者。由于推出了基于DTG的通用方案，并将其纳入 资源有限国家国民待遇指南，短短5年内，1500万艾滋病毒-1感染者 人们将接受DTG治疗。这包括育龄妇女，她们仍然是严重的感染者。 人口(艾滋病规划署数据，2020年)。然而，近年来，来自不同临床和前期的越来越多的数据 临床研究表明，DTG与出生缺陷和出生后发育有关 神经学异常。因此，出现了对孕妇或孕妇使用基于DTG的方案的担忧 育龄妇女。知识差距：虽然临床队列研究确定了神经学风险 宫内DTG暴露后婴儿的异常，DTG相关的潜在机制 发育神经毒性，特别是出生时无结构、脑或脊髓畸形的婴儿 仍然不为人知。我们的初步数据：DTG被发现是一种MMPs的广谱抑制剂。这种药 被发现与锌离子结合在催化区域，导致MMPs活性的抑制。此外，在中国的研究 怀孕的小鼠显示DTG可以穿过胎盘屏障，在胎儿中枢神经系统蓄积 并抑制胎脑发育关键期的MMPs活性。进一步的产后评估 宫内DTG暴露对小鼠大脑健康的影响确定了神经炎症和神经元 损坏。这些数据表明DTG对MMPs活性的异常调节在 妊娠和随之而来的神经毒性。假设：我们假设DTG抑制MMPs活性 在怀孕期间会损害神经发育。研究策略：(1)测定剂量依赖酶 不同INSTI类药物DTG、比替格列韦和卡替格列韦的抑制动力学 (驾驶室)。(2)检测DTG对胚胎MMPs功能和表达的纵向剂量依赖性影响 妊娠期中枢神经系统。(3)明确DTG抑制MMPs对胚胎神经发育的影响 妊娠期间的生理过程(血管生成和神经生成)。(4)评估BIC和CAB是否 也通过相同的机制影响体内MMPs的活性和相关的神经发育过程 以评估班级效应。研究团队：我们组建了一支多学科团队，具备以下专业知识 神经病理学，免疫学，药剂学，病毒学和生物成像，以评估宫内DTG和 来自INSTI类暴露的其他药物对胚胎大脑发育的影响。结果：成功的结果 该建议将极大地促进该领域对DTG抑制MMPs活性的理论的理解 作为DTG相关发育神经毒性的潜在机制，以及是否 可以将相同的机制链接到INSTI类的其他成员。

Project Summary Background: Dolutegravir (DTG), an integrase strand transfer inhibitor (INSTI), currently recommended as both, first-line therapy and part of either switching or salvage regimens for the treatment of human immunodeficiency virus type-1 (HIV-1) infected patients. Due to the roll out of generic DTG-based regimen and its inclusion in national treatment guidelines in resource limited countries (RLCs), in just 5 years, 15 million HIV-1 infected people will be treated with DTG. This includes women of child-bearing age, who remain a significant infected population (UNAIDS data, 2020). However, during recent years, growing data from different clinical and pre- clinical studies have suggested that the DTG is associated with birth defects and with postnatal developmental neurologic abnormalities. Thus, concerns emerged for the usage of DTG-based regimens in pregnant women or those of child-bearing age. Knowledge gap: While clinical cohort studies identified risk of neurologic abnormalities in babies following in utero DTG-exposure, underlying mechanism for DTG-associated developmental neurotoxicity, particularly in babies born without structural, brain or spinal cord, malformations remains unknown. Our preliminary data: DTG was found to be a broad-spectrum inhibitor of MMPs. The drug was found to bind Zn++ at the catalytic domain, leading to inhibition of MMPs activities. Moreover, studies in pregnant mice showed that DTG can cross the placental barrier, accumulate in the fetal central nervous system (CNS) and inhibit MMPs activity during the critical period of fetal brain development. Further postnatal evaluation of brain health in mice pups following in utero DTG exposures identified neuroinflammation and neuronal damage. These data demonstrated an association between DTG dysregulation of MMPs activities during gestation and consequent neurotoxicity. Hypothesis: We hypothesize that DTG inhibition of MMPs activities during gestation impairs neurodevelopment. Research Strategy: (1) Determine dose-dependent enzyme inhibition kinetics of different drugs from INSTI class, which include DTG, bictegravir (BIC) and cabotegravir (CAB). (2) Determine longitudinal dose-dependent affect of DTG on MMPs function and expression in embryo CNS during gestation. (3) Identify the impact of MMPs inhibition by DTG on embryo neurodevelopmental physiological processes (angiogenesis and neurogenesis) during pregnancy. (4) Evaluate whether BIC and CAB also affect the MMPs activities and associated neurodevelopmental processes in vivo through same mechanism to assess the class effect. Research Team: We have assembled a multidisciplinary team with expertise in neuropathology, immunology, pharmaceutics, virology, and bioimaging to evaluate impact of in utero DTG and of other drugs from INSTI class exposure on the embryo brain development. Outcome: Successful outcome of the proposal will significantly advance the field in understanding the theory of DTG inhibition of MMPs activities during gestation as the underlying mechanism for DTG-associated developmental neurotoxicity and whether the same mechanism can be linked to other members of the INSTI class.