Maternal immune activation remodeling of offspring glycosaminoglycan sulfation patterns during neurodevelopment

摘要 产前或产后发育期间的母体免疫激活（MIA）显著增加了以下风险： 后代神经发育障碍（NDD）。越来越多的证据表明， MIA刺激（感染或环境），后代表现出终身神经病理学缺陷的风险增加 从脑容量的减少到神经回路组织的改变。大脑细胞外基质- 含有软骨素和硫酸皮肤素的糖胺聚糖（CS/DS-GAG）是脑内 发育，并可通过神经免疫反应进行生化改变。CS/DS-GAG丰度缺陷 和/或硫酸化图案化（4S（CS-A）、2S 4S（CS-B/DS）、6S（CS-C）、2S 6S（CS-D）、4S 6S（CS-E）、0 S（CS-O）） 导致与受MIA影响的后代报告的类似神经精神行为的表现，但 MIA是否以及如何影响后代的脑基质尚不清楚。通过采用一种新颖的激光捕获 显微切割耦合质谱法（LMD-LC-MS/MS），我们的初步数据提供了 第一个证据表明，整个研究期间CS/DS-GAG硫酸化模式差异存在区域间和区域内差异。 小鼠和非人类灵长类动物（NHP）大脑的发育。具体来说，海马体表现出显著的 与皮质相比，发育中的6S（CS-C）和2S 6S（CS-D）异构体均增加，这意味着 海马在邻近区域成熟后很长时间内仍保持发育可塑性。此外，我们显示 NHP妊娠期间感染性寨卡病毒MIA降低了发育中2S 6S的丰度， （CS-D）轴突生长因子引诱异构体在海马，表明发育不良的神经回路形成 感染性MIA后，而非感染性母体高脂饮食（mHFD）MIA在哺乳期小鼠 降低海马中发育6S（CS-C）可塑性异构体的丰度，表明 加速海马神经回路对非感染性MIA的早期成熟。言下之意 感染性和非感染性MIA损伤均影响脑CS/DS-GAG的时空调节， 硫酸化模式符合一个全球性的相互联系的理论，该理论将一系列MIA损伤与后代大脑的变化联系起来 通过重新编码CS/DS-GAG来促进神经发育。从这些结果，我们建议1）确定如何MIA 暴露影响后代CS/DS-GAG的时空表达，并将这些变化与NDD联系起来。 2）机械地研究这些MIA诱导的后代CS/DS-GAG的变化如何影响聚糖， 参与神经发育的蛋白质相互作用，以及3）设计最先进的纳米孔测序 能够对生物CS/DS-GAG进行单分子测序以发现聚糖-蛋白质结合的技术 元素这一多学科的建议具有重要的转化潜力，以澄清MIA暴露如何导致 通过儿童期CS/DS-GAG硫酸化模式的变化与神经精神疾病 神经发育，并提供了预防和治疗精神健康疾病的有价值的目标。

ABSTRACT Maternal immune activation (MIA) during prenatal or postnatal development significantly increases the risk for offspring neurodevelopmental disorders (NDDs) later in life. Growing evidence suggest that regardless of the MIA stimuli (infectious or environmental), offspring exhibit an enhanced risk for lifelong neuropathology defects ranging from reduced brain volume to alterations in neurocircuit organization. The brain extracellular matrix- containing chondroitin and dermatan sulfate-glycosaminoglycans (CS/DS-GAGs) are key regulators of brain development and can be biochemically altered by neuroimmune responses. Defects in CS/DS-GAG abundance and/or sulfation patterning (4S (CS-A), 2S4S (CS-B/DS), 6S (CS-C), 2S6S (CS-D), 4S6S (CS-E), 0S (CS-O)) result in the manifestation of similar neuropsychiatric behaviors as reported in offspring affected by MIA, but whether and how MIA affects offspring brain matrix is unknown. By employing a novel laser capture microdissection coupled mass spectrometry methodology (LMD-LC-MS/MS), our Preliminary Data provide the first evidence for inter- and intra-regional differences in CS/DS-GAG sulfation pattern differences throughout the developing mouse and non-human primate (NHP) brain. Specifically, the hippocampus exhibits a significant increase in both developmental 6S (CS-C) and 2S6S (CS-D) isomers compared to the cortex, implying that the hippocampus remains developmentally plastic long after the maturation of adjacent regions. Moreover, we show that infectious Zika virus MIA during gestation in NHPs decreases the abundance of the developmental 2S6S (CS-D) axonal growth factor attractant isomer in the hippocampus, suggesting stunted neurocircuit formation after infectious MIA, while the non-infectious maternal high fat diet (mHFD) MIA during lactation in mice decreases the abundance of the developmental 6S (CS-C) plasticity isomer in the hippocampus, suggested accelerated early maturation of hippocampal neurocircuits in response to non-infectious MIA. The implication that both infectious and non-infectious MIA insults influence the spatiotemporal regulation of brain CS/DS-GAG sulfation patterns fits a global interconnecting theory linking a range of MIA insults with changes in offspring brain neurodevelopment through re-coding of CS/DS-GAGs. From these results, we propose to 1) determine how MIA exposure affects spatiotemporal expression of offspring CS/DS-GAGs and link these changes to NDDs later in life, 2) mechanistically investigate how these MIA-induced changes in offspring CS/DS-GAGs influence glycan- protein interactions involved in neurodevelopment, and 3) engineer a state-of-the-art nanopore sequencing technology capable of single-molecule sequencing of biological CS/DS-GAGs to discover glycan-protein binding elements. This multidisciplinary proposal has important translational potential to clarify how MIA exposure leads to neuropsychiatric illness through changes in CS/DS-GAG sulfation patterning during childhood neurodevelopment and provides valuable targets in the prevention and treatment of mental health diseases.