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).

项目概要/摘要： 如果在发育过程中感染弓形虫，可能会对发育中的胎儿造成毁灭性后果。 每年都会发生大量先天性弓形虫感染（美国约 4000 起）。 全球有 200,000 例），使弓形虫成为最重要的致畸病原体之一。 预测未接触过弓形虫的孕妇的感染是否最终会传染给胎儿。 弓形虫病仅在弓形虫突破胎盘屏障后发生，但人们对细胞的了解甚少 以及宿主与病原体相互作用期间发生的分子事件，以及这些事件如何影响感染 在本提案中，我们概述了一系列补充实验，旨在了解结果。 当弓形虫遇到胎盘细胞时会发生这种情况，以及这可能如何影响感染 我们工作的一个主要重点是构成初级滋养层的胎盘滋养层。 我们广泛的初步和已发表的数据坚定地确定了胎儿发育和母体血液之间的界面。 确定胎盘合体滋养层 (SYN) 但细胞滋养层 (CYT) 不抵抗弓形虫 感染，并且滋养细胞的弓形虫感染诱导独特的转录反应 与迄今为止研究的大多数细胞类型相比，在目标 1 中，我们利用了多种遗传上易于处理的 SYN 模型。 功能和开发来识别 SYN 抗性机制并在分子水平上验证它们。 这一目标的意义源于这样一个事实：SYN 是迄今为止唯一研究的细胞类型。 在目标 2 中，我们强调了先天免疫信号传导的重要性。 在 Subaim 2.2 中，我们将通过量化来做到这一点。 一组独特的人血清样本中的多种免疫调节细胞因子，纵向覆盖 怀孕期间的妊娠血清转换事件，然后使用多变量分析将免疫联系起来 在 Subaim 2.2 中，我们将从基因角度消除多个宿主靶向。 弓形虫中的效应器（包括特别改变胎盘细胞免疫调节景观的效应器）， 并将先天性传播动力学与成熟小鼠的免疫调节景观联系起来 先天性传播模型的影响来自于（1）原发性胎盘的使用。 CYT 和 SYN 的组织和易处理细胞系模型，用于解码关键的耐药分子机制 和敏感性，（2）使用弓形虫感染之前、期间和之后的血清样本来稳健量化 先天性暴露事件期间宿主反应的变化以及（3）协同且公认的 共同 PI 团队（Coyne 和 Boyle）的合作，利用他们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（博伊尔）以及胎盘发育的细胞和分子基础 和病原体抗性（Coyne）。