Placental resistance and response to the teratogenic pathogen Toxoplasma gondii

胎盘对致畸病原体弓形虫的抵抗力和反应

• 批准号: 10453973
• 负责人: JON P BOYLE
• 金额: $ 58.2万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10453973
• 关键词: Acute; Address; Adhesions; Adopted; Affect; Antiparasitic Agents; Biological Assay; Biological Markers; Biology; Blindness; Blood; Bypass; Cell Line; Cells; Characteristics; Clinical; Collaborations; Congenital Abnormality; Congenital Toxoplasmosis; Cytokine Signaling; Data; Defense Mechanisms; Development; Developmental Delay Disorders; Event; Fetal Development; Fetal health; Fetus; Gene Expression Profile; Genetic Transcription; Goals; Health; Human; Immune; Immune response; Immune signaling; Infection; Knowledge; Lead; Link; Mammalian Cell; Maternal Health; Maternal-Fetal Exchange; Mediating; Modeling; Molecular; Molecular Biology; Multivariate Analysis; Outcome; Parasites; Physiological; Placenta; Placentation; Predisposition; Pregnancy; Pregnancy Outcome; Pregnancy loss; Pregnant Women; Production; Proteins; Proteoglycan; Publishing; Research Design; Resistance; Sampling; Series; Serum; Signal Pathway; Signal Transduction; Spontaneous abortion; Sulfate; Surface; Syncytiotrophoblast; Teratogens; Testing; Tissues; Toxoplasma; Toxoplasma gondii; Toxoplasmosis; Treatment Protocols; Vertical Disease Transmission; Woman; Work; base; cell type; chemokine; cytokine; cytotrophoblast; design; diagnostic tool; enhancing factor; experimental study; fetal; human pathogen; immunoregulation; in utero; in vitro Model; in vivo; infection risk; mouse model; mutant; neonate; pathogen; pathogen exposure; placental infection; pregnant; resistance mechanism; response; seroconversion; transcriptome; transmission process; trophoblast

PROJECT SUMMARY/ABSTRACT: Toxoplasma gondii can have devastating consequences in the developing fetus if it is acquired during pregnancy. A significant number of congenital T. gondii infections occur yearly (~4000 in the U.S. and over 200,000 worldwide), making T. gondii among the most important teratogenic pathogens. To date it is not possible to predict whether infection of a T. gondii-naïve pregnant woman will ultimately transmit to the fetus. Congenital toxoplasmosis occurs only after T. gondii breaches the placental barrier, yet very little is known about the cellular and molecular events that occur during this host-pathogen interaction, nor how these events affect infection outcome. In this proposal we outline a series of complementary experiments aimed at understanding what happens when Toxoplasma gondii encounters cells of the placenta and how this might impact infection outcome in the developing fetus. A major focus of our work is on placental trophoblasts which form the primary interface between the developing fetus and maternal blood. Our extensive preliminary and published data firmly establish that placental syncytiotrophoblasts (SYNs) but not cytotrophoblasts (CYTs) resist Toxoplasma infection, and that Toxoplasma infection of trophoblasts induces a transcriptional response that is unique compared to most cell types studied to date. In Aim 1 we exploit multiple genetically tractable models of SYN function and development to identify SYN resistance mechanisms and validate them at the molecular level. Significance of this aim derives from the fact that SYNs are unique in being the only cell type studied to date that is intrinsically resistant to T. gondii infection. In Aim 2 we address the importance of innate immune signaling at the maternofetal interface during congenital transmission. In Subaim 2.2. we will do this by quantifying multiple immunomodulatory cytokines in a unique set of human serum samples that longitudinally cover gestational seroconversion events during pregnancy, and then use multivariate analyses to link immune signaling profiles to different infection outcomes. In Subaim 2.2. we will genetically ablate multiple host-targeting effectors in T. gondii (including one that alters the immunoregulatory landscape specifically in placental cells), and link congenital transmission dynamics to the immunomodulatory landscape in a well-established mouse model of congenital transmission. Impact of the proposed studies derives from (1) the use of primary placental tissues and tractable cell line models of CYTs and SYNs to decode critical molecular mechanisms of resistance and susceptibility, (2) the use of serum samples before, during and after T. gondii infection to robustly quantify changes in the host response during congenital exposure events and (3) the synergistic and well-established collaboration of the co-PI team (Coyne and Boyle) which leverages their expertise in the molecular biology of Toxoplasma host-pathogen interactions (Boyle) and the cellular and molecular basis of placental development and pathogen resistance (Coyne).

项目摘要/摘要： 如果弓形虫在发育中的胎儿在发育室中获得毁灭性后果 孕。每年发生的大量先天性T. gondii感染（在美国及以上约4000个） 在全球范围内200,000），使T. gondii成为最重要的致畸病原体。迄今为止不可能 为了预测gondii孕妇的感染是否最终会传播给胎儿。 弓形虫病仅在T. gondii打破斑点屏障之后才发生，但对细胞知之甚少 以及在这种宿主病原体相互作用期间发生的分子事件，也不是这些事件如何影响感染 结果。在此提案中，我们概述了一系列旨在了解什么的完整实验 发生弓形虫弓形体遇到斑块的细胞以及这可能影响感染时发生的发生 发展中的胎儿的结果。我们工作的主要重点是构成主要的滋养细胞 发育中的胎儿和孕妇血之间的接口。我们广泛的初步和发布数据首先 确定胎盘突触植物细胞（Syns），但不能抵抗弓形虫的细胞增多质细胞（CYT） 感染，滋养细胞的弓形虫感染引起了独特的转录反应 与迄今为止研究的大多数细胞类型相比。在AIM 1中，我们利用了多个SYN的多个通用典型模型 功能和发育以识别syn抗性机制并在分子水平上验证它们。 此目标的意义源于以下事实：Syns是迄今为止唯一的单元格类型。 这在内在耐药性抗gondii感染。在AIM 2中，我们解决了先天免疫信号的重要性 在先天性传播期间的Maternofetal界面。在Subaim 2.2中。我们将通过量化来做到这一点 多种免疫调节细胞因子在一组纵向覆盖的独特人血清样品中 怀孕期间的妊娠血清转化事件，然后使用多元分析来连接免疫 对不同感染结果的信号谱。在Subaim 2.2中。我们将一般烧毁多个主机靶向 在T. gondii中的影响（包括一种特定在斑点细胞中的免疫调节景观的影响）， 并将先天性传输动力学与公认的鼠标中的免疫调节景观联系起来 先天性传播的模型。拟议研究的影响源于（1）使用原发性位置 组织和可拖动的细胞系模型和Syns的抗药性临界分子机制解码 （2）在T. gondii感染之前，之中和之后使用血清样品来稳健量化 先天性暴露事件期间宿主反应的变化以及（3）协同且建立良好的 Co-Pi团队（Coyne and Boyle）的合作，该团队利用其在分子生物学方面的专业知识 弓形虫宿主 - 病原体相互作用（Boyle）以及斑点发育的细胞和分子基础 和病原体抗性（Coyne）。