HCMV infection of human placental trophoblast and hematopoietic progenitors

人胎盘滋养层和造血祖细胞的 HCMV 感染

• 批准号: 8535904
• 负责人: LENORE PALMA PEREIRA
• 金额: $ 54.6万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535904
• 关键词: Address; Affect; Antibodies; Area; Birth; Blood; CD34 gene; Cell Cycle; Cell Cycle Progression; Cell Differentiation process; Cell Lineage; Cells; Characteristics; Chorionic villi; Chromosomes; Commit; Complex; Congenital Abnormality; Cytomegalovirus; Cytomegalovirus Infections; Cytoplasm; DNA; Development; Disease; Embryo; Employee Strikes; Environment; Epithelial; Equilibrium; Erythrocytes; Erythroid; Fibrosis; First Pregnancy Trimester; Geminin; Goals; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Hypoxia; Infection; Infection prevention; Infectious Agent; Injury; Lead; Licensing; Link; Lymphoid; Measures; Modeling; Molecular; Mus; Myelogenous; Myeloid Cells; Neuroglia; Nuclear Protein; Organ; Pathogenesis; Placenta; Placentation; Play; Population; Pregnancy; Prevention strategy; Proteins; Risk; Role; S Phase; Somatic Cell; Source; Stem cells; Surface; Syncytiotrophoblast; Testing; Tropism; Uterus; Villous; Viral; Virion; Virus; Woman; Work; base; congenital infection; cytotrophoblast; embryo/fetus; fetal; in utero; insight; macrophage; neutralizing monoclonal antibodies; novel; novel strategies; pathogenic bacteria; placental stem cell; pluripotency; prevent; progenitor; protein expression; research study; self-renewal; stem cell differentiation; stem cell fate specification; stem cell population; theories; transmission process; trophoblast

DESCRIPTION (provided by applicant): Mechanisms of transplacental transmission of human cytomegalovirus (HCMV), the leading viral cause of congenital infection affecting 1-3% of births in the U.S., are unknown. Primary maternal infection in first trimester poses a 40-50% risk of transplacental transmission and permanent birth defects in 15% of diseased babies. We propose to study HCMV infection of the human placental progenitors of the trophoblast (TB) and hematopoietic lineages, and the consequences in terms of the molecular mechanisms that dysregulate development. The Fisher group identified the early-gestation human placental stroma as a niche for cells that co-expressed markers of pluripotency and determinants of mouse TB fate, which suggested that this region is one source of human TB progenitor cells (TBPCs). They isolated these cells and developed lines of continuously self-renewing TBPCs. When cultured under conditions that triggered TB differentiation, the cells formed the mature human TB populations-multi-nucleated, transport syncytiotrophoblasts (STBs) and invasive CTBs. In the same region, the Fisher group identified hematopoietic stem cells (HSCs) as well intermediate precursors of the myeloid- and erythroid-committed lineages, suggesting active hematopoiesis. Cultured in defined medium, the CD34++CD45low HSCs contributed to erythrocytes and myeloid cells. Together these studies showed that the human placenta is a source of TBPCs that populate the chorionic villi and HSCs that might play a role in development of the hematopoietic system of the embryo/fetus. It is likely that placental HSCs also contribute to the placental Hofbauer macrophage population. TBPCs are highly susceptible to infection with a pathogenic HCMV strain that induced expression of proteins controlling cell-cycle progression and the balance between self-renewal and lineage-commitment. Upregulated molecules were mislocalized to the cytoplasm and accumulated in the virion assembly compartment. Additional experiments showed that HCMV infected placentally-derived HSCs. Here, we propose testing the theory that HCMV infection alters the balance between TBPC self-renewal and differentiation (Aim 1). We also hypothesize that infection of HSCs impairs their ability to form myeloid- and erythroid-committed lineages (Aim 2). These studies will provide new insights into viral effects on differentiation of TBPCs, which carry out the specialized functions of the placenta, and HSCs, which contribute to fetal hematopoiesis and the placental immunological barrier. Then we will use this information to determine if the same phenotypic changes are observed in placentas that were congenitally infected in utero. We will also measure the ability of human neutralizing MoAbs to protect chorionic villi from HCMV infection (Aim 3). In summary, these studies will provide new information about the effects of HCMV on TBPC and HSC self-renewal and differentiation. We also expect to gain insights into novel approaches for preventing HCMV transmission and the attendant birth defects.

描述（由申请人提供）：人类巨细胞病毒 (HCMV) 经胎盘传播的机制尚不清楚，HCMV 是影响美国 1-3% 新生儿的先天性感染的主要病毒原因。妊娠早期的原发性母体感染造成 40-50% 的经胎盘传播风险，并导致 15% 的患病婴儿出现永久性出生缺陷。我们建议研究滋养层 (TB) 和造血谱系的人胎盘祖细胞的 HCMV 感染，以及发育失调的分子机制方面的后果。 Fisher小组发现，妊娠早期的人胎盘基质是共表达多能性标记和小鼠结核病命运决定因素的细胞的生态位，这表明该区域是人类结核祖细胞（TBPC）的来源之一。他们分离了这些细胞并开发了不断自我更新的 TBPC 系。当在引发结核分化的条件下培养时，这些细胞形成了成熟的人类结核群体——多核、运输合体滋养层（STB）和侵袭性CTB。在同一区域，费舍尔研究小组发现了造血干细胞（HSC）以及骨髓和红系定型谱系的中间前体，这表明造血活动活跃。在确定成分培养基中培养时，CD34++CD45low HSC 产生红细胞和骨髓细胞。这些研究共同表明，人类胎盘是绒毛膜绒毛中的 TBPC 和 HSC 的来源，它们可能在胚胎/胎儿造血系统的发育中发挥作用。胎盘 HSC 也可能对胎盘 Hofbauer 巨噬细胞群有贡献。 TBPC 非常容易受到致病性 HCMV 菌株的感染，该菌株诱导控制细胞周期进程以及自我更新和谱系承诺之间平衡的蛋白质表达。上调的分子被错误定位到细胞质并在病毒粒子组装室中积累。其他实验表明 HCMV 感染胎盘来源的 HSC。在这里，我们建议测试 HCMV 感染改变 TBPC 自我更新和分化之间的平衡的理论（目标 1）。我们还假设 HSC 的感染会损害它们形成髓系和红系定向谱系的能力（目标 2）。这些研究将为病毒对 TBPC（执行胎盘的特殊功能）和 HSC（有助于胎儿造血和胎盘免疫屏障）分化的影响提供新的见解。然后我们将利用这些信息来确定在子宫内先天性感染的胎盘中是否观察到相同的表型变化。我们还将测量人类中和 MoAb 保护绒毛膜绒毛免受 HCMV 感染的能力（目标 3）。总之，这些研究将提供关于 HCMV 对 TBPC 和 HSC 自我更新和分化影响的新信息。我们还希望深入了解预防 HCMV 传播和随之而来的出生缺陷的新方法。