Transcriptomics and biomarkers of fetal neuroinflammation

胎儿神经炎症的转录组学和生物标志物

基本信息

批准号：
10175560
负责人：
SUHAS KALLAPUR
金额：
$ 26.74万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10175560
关键词：
Administrative Supplement Affect Alzheimer&apos s Disease Animals Anti-Inflammatory Agents Antibiotics Archives Area Bioinformatics Biological Markers Brain Brain Injuries Brain region Budgets Cell Nucleus Cells Cerebral Palsy Child Childhood Clinical Data Data Data Set Development Digestion Disease Drug Targeting Escherichia coli Exposure to Fetal Membranes Fetus Freezing Funding Gene Expression Gene Expression Profiling Goals Grant Human Immunology Inflammation Injections Injury Interleukin-1 Interleukin-1 Receptors Knowledge Lactation Lipopolysaccharides Liquid substance Macaca mulatta Maps Mediating Medical Microglia Modeling Molecular Mucous Membrane National Institute of Child Health and Human Development Neonatal Neurodegenerative Disorders Neurologic Neurosciences Nitrogen Nuclear Nuclear RNA Pathogenesis Peptide Hydrolases Perinatal Pharmaceutical Preparations Pre-Clinical Model Prefrontal Cortex Pregnancy Pregnant Women Premature Labor Public Health Recombinants Research Activity Resources Rhesus Risk Saline Sampling Signal Transduction Subcutaneous Injections Testing Time Treatment Efficacy Woman adverse pregnancy outcome anakinra animal resource animal tissue autism spectrum disorder candidate marker cost developmental neurobiology drug efficacy efficacy study fetal gene discovery genome-wide histological studies insight intrauterine infection mRNA sequencing molecular targeted therapies neonate neurodevelopment neuroinflammation newborn brain injury nonhuman primate novel parent grant pharmacokinetics and pharmacodynamics potential biomarker pre-clinical preclinical development preclinical study pregnant prenatal prenatal exposure response single cell sequencing single-cell RNA sequencing therapeutic candidate transcriptome sequencing transcriptomics verification and validation white matter

项目摘要

Summary This is an administrative supplement proposal to the parent grant HD98389 (2019-24) titled “Anti-inflammatory drug target to reduce adverse pregnancy outcomes”. The goal of the parent grant is to develop Anakinra (recombinant human IL1receptor antagonist) as a safe and effective therapeutic for intrauterine infection/inflammation mediated preterm labor and fetal inflammation. In the parent grant, pregnant Rhesus macaques will be given Anakinra in a model of intrauterine inflammation (IUI). We will now leverage the resources of the parent grant to expand the Aims of the parent grant. Fetal neuro-inflammation is associated with later development of cerebral palsy (CP) and autism spectrum disorders (ASD) with significant medical and public health implications. As part of the parent grant activities, we have demonstrated that inhibition of IL1 signaling using Anakinra significantly decreased fetal inflammation induced by intraamniotic injection of E. coli LPS. We have now developed capability of extracting intact nuclei from fetal frozen brain sample and to do single cell sequencing. Nuclear as compared to cytosolic RNA seq has the advantage of not needing protease digestion at 37oC thereby eliminating artifactual gene expression. Furthermore, nuclear RNA seq can be done in archived frozen brain sample thereby sparing precious animal use and reducing costs. In this administrative supplement grant, we will test the hypothesis that inhibition of IL1 signaling by Anakinra will decrease region specific fetal neuro inflammation induced by intraamniotic LPS. We propose doing single nuclear mRNA seq from 2 regions of the brain – the peri-ventricular white matter, and the prefrontal cortex – regions implicated in the pathogenesis of CP and ASD. We will compare region specific single nuclear mRNA seq in three groups of Rhesus macaque fetuses at about 80% gestation: 1) Controls – intraamniotic injection (IA) of saline, 2) IA LPS, 3) IA LPS + Anakinra. All exposures are for 16h, a time point that we previously demonstrated to be optimum for the proposed studies. The animal resources funded by the parent grant are already available. Thus, single cell transcriptomic studies will be greatly facilitated and feasible within the year of funding. This fetal brain transcriptomic study will reveal mechanistic insights in the pathogenesis of neuroinflammation resulting from IUI, and identify leads for biomarker sets for CP and ASD. The study will also further assist in pre-clinical development of Anakinra as a therapeutic candidate for IUI.

概括这是对家长补助金 HD98389 (2019-24) 题为“抗炎”的行政补充提案减少不良妊娠结局的药物目标”。母基金的目标是开发 Anakinra。（重组人 IL1 受体拮抗剂）作为一种安全有效的宫内治疗药物感染/炎症介导的早产和胎儿炎症在母体中，怀孕的恒河猴。我们将在宫内炎症 (IUI) 模型中给猕猴注射阿那白滞素。母体补助金的资源扩大与胎儿神经炎症相关。后来发展为脑瘫（CP）和自闭症谱系障碍（ASD），具有重大的医学和作为家长资助活动的一部分，我们已经证明了 IL1 的抑制作用。使用阿那白滞素（Anakinra）的信号显着降低羊膜内注射大肠杆菌引起的胎儿炎症 LPS。我们现在已经开发出了从胎儿冷冻脑样本中提取完整细胞核并进行单次分离的能力。细胞核测序与胞质 RNA 测序相比，具有不需要蛋白酶消化的优点。在 37oC 下消除人工基因表达此外，核 RNA 测序可以在存档中进行。冷冻大脑样本，从而节省宝贵的动物使用并降低成本。授予，我们将测试以下假设：Anakinra 抑制 IL1 信号传导会降低区域特异性我们建议对 2 进行单核 mRNA 测序。大脑区域——脑室周围白质和前额皮质——与我们将比较三组中的区域特异性单核 mRNA seq。约 80% 妊娠的恒河猴胎儿：1) 对照 – 羊膜内注射 (IA) 生理盐水，2) IA LPS， 3) IA LPS + Anakinra 所有暴露时间均为 16 小时，这是我们之前证明的最佳时间点。拟议的研究。由家长资助的动物资源已经可用。在资助的一年内，转录组研究将得到极大的便利和可行。转录组学研究将揭示 IUI 引起的神经炎症发病机制的机制见解，并确定 CP 和 ASD 生物标志物组的线索该研究还将进一步协助临床前开发。阿那白滞素 (Anakinra) 作为 IUI 的候选治疗药物。