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

项目摘要 背景：Dolutegravir（DTG）是一种整合酶链转移抑制剂（IGI），目前被推荐作为两种药物， 人类免疫缺陷治疗的一线治疗和部分转换或挽救治疗方案 1型病毒（HIV-1）感染患者。由于推出了基于DTG的通用方案，并将其纳入 在资源有限的国家（RLC）的国家治疗指南，在短短5年内，1500万HIV-1感染者 人们将接受DTG治疗。这包括育龄妇女，她们仍然是受感染的主要群体。 人口（UNAIDS数据，2020年）。然而，近年来，来自不同临床和预处理的数据越来越多， 临床研究表明，DTG与出生缺陷和出生后发育有关， 神经系统异常因此，出现了对孕妇使用基于DTG的方案的担忧， 生育年龄的人。知识差距：虽然临床队列研究确定了神经系统疾病的风险， 胎儿宫内DTG暴露后的异常，DTG相关的潜在机制 发育神经毒性，特别是在出生时没有结构、大脑或脊髓畸形的婴儿中 仍然未知。我们的初步数据：DTG被发现是一种广谱的MMPs抑制剂。药物 发现在催化结构域结合Zn++，导致MMPs活性的抑制。此外，研究 孕鼠实验表明，DTG可穿过胎盘屏障，在胎鼠中枢神经系统内蓄积 (CNS)在胎儿脑发育的关键时期抑制MMPs活性。产后进一步评估 在子宫内DTG暴露后， 损害这些数据表明，DTG失调的MMPs活动之间的关联， 妊娠和随后的神经毒性。假设：我们假设DTG抑制MMPs活性 会损害神经发育研究策略：（1）确定剂量依赖性酶 不同药物的抑制动力学，包括DTG，bictegravir（BIC）和cabotegravir （CAB）。(2)确定DTG对胚胎中MMPs功能和表达的纵向剂量依赖性影响 中枢神经系统在妊娠期间。(3)确定DTG抑制MMPs对胚胎神经发育的影响 生理过程（血管生成和神经发生）。(4)评估BIC和CAB是否 也通过相同的机制影响MMPs的活性和相关的神经发育过程 来评估阶级效应。研究团队：我们组建了一个多学科团队，具有以下方面的专业知识： 神经病理学、免疫学、药学、病毒学和生物成像，以评价子宫内DTG的影响， 其他类药物暴露对胚胎脑发育的影响。结果：成功的结果 该建议将显著推进该领域对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 同样的机制可以链接到其他成员的EMAI类。

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.