VTCN1 regulation of MHC in early human placental development

VTCN1 对人类早期胎盘发育中 MHC 的调节

• 批准号: 10092932
• 负责人: Toshihiko Ezashi
• 金额: $ 19.38万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-31 至 2022-07-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092932
• 关键词: Abbreviations; Abnormal placentation; Address; Affect; Aggressive behavior; Allogenic; Allografting; BMP4; Binding; Blood group antigen f; Cell Proliferation; Cell model; Cell surface; Cells; Cervix Neoplasms; Characteristics; Clinical; Complex; DNA Binding Domain; Decidua; Detection; Development; Disease; Dissection; Down-Regulation; Endometrial Neoplasms; Ensure; Environment; Ethics; Event; Fetal Development; Fetal Growth Retardation; Fetus; First Pregnancy Trimester; Genetic Transcription; HLA Antigens; Hepatitis B e Antigens; Histocompatibility Antigens; Histocompatibility Antigens Class II; Human; Human Chorionic Gonadotropin; Immune; Immunocompetent; Immunohistochemistry; Immunologics; Immunology; Implant; In Vitro; Invaded; Knowledge; Learning; Logistics; MAP Kinase Gene; MHC class II transactivator protein; Major Histocompatibility Complex; Malignant Neoplasms; Maternal-Fetal Exchange; Messenger RNA; Mitogen-Activated Protein Kinases; Modeling; Mothers; Normal tissue morphology; Nucleotides; Pathway interactions; Pattern; Physiological; Placenta; Placentation; Play; Post-Transcriptional Regulation; Pre-Eclampsia; Pregnancy; Pregnancy Rate; Pregnancy loss; Premature Birth; Process; Prognosis; Proteins; Regimen; Regulation; Regulatory Element; Reproductive Immunology; Research; Risk; Role; SET Domain; Sampling; Second Pregnancy Trimester; Small Interfering RNA; Surface; Syncytiotrophoblast; T-Cell Activation; T-Cell Receptor; TCR Activation; Testing; Tissues; Trans-Activators; Undifferentiated; VTCN1 gene; Vascularization; beta-2 Microglobulin; cancer cell; cytotrophoblast; early pregnancy; early pregnancy loss; embryonic stem cell; healthy pregnancy; human embryonic stem cell; human tissue; implantation; improved; in vitro Model; induced pluripotent stem cell; inhibitor/antagonist; innovation; knock-down; matrigel; member; neovascularization; novel; novel diagnostics; novel therapeutics; ovarian neoplasm; overexpression; pancreatic neoplasm; pathogen; pregnancy disorder; programmed cell death ligand 1; protein expression; receptor; response; selective expression; stem cell differentiation; stem cell model; stem cells; tool; trophoblast; tumor; tumor immunology; tumor-immune system interactions

Improper peri-implantation placental development may contribute to feto-maternal diseases (preterm birth, intrauterine growth retardation, preeclampsia) and high rates of pregnancy loss. Since these events occur prior to clinical detection of pregnancy, studies of very early human placental development are ethically and logistically constrained. The proposed project uses an innovative human embryonic stem cell (hESC)-derived model to study primitive trophoblast in vitro. The placenta resembles a cancer in that an implanting fetus and a developing cancer both involve host tissue invasion, massive cellular proliferation, neo-vascularization, and alterations in the immediate immune microenvironment. The placenta of the implanting fetus is one of the only non-cancerous tissues that displays physiologic downregulation of the classical major histocompatibility complex (MHC) class I (MHC-I) transplantation antigens, human leukocyte antigens (HLA)-A and -B. While positive regulatory elements for MHC class II regulation are described, little is known about such regulation of MHC class I expression in noncancerous tissues, including the placenta. Human embryonic stem cells (hESCs) have been used in a novel in vitro model of the stages of human placental development that occur prior to clinical detection of pregnancy. Several mRNAs and proteins that are selectively expressed at high levels in early pregnancy have been identified by using the model. Of those studied, all appear to play roles in invasion. Remarkably, siRNA silencing of one of these proteins, V-Set Domain Containing T Cell Activation Inhibitor 1 (VTCN1), in our model for early trophoblast strongly up-regulated the expression of the MHC class I human leukocyte antigens (HLA)-A and -B, which are normally not expressed in human trophoblast, slightly up-regulated the normally resident HLA-C, but had no effect on expression of the placenta-specific HLA-G. VTCN1 is recognized as a member of the B7 superfamily of cell-surface co-receptors that regulate T cell receptor interactions with MHC molecules. It has been postulated that VTCN1 neo-expression affects cancer cells’ ability to evade host immune detection. These results highlight the efficacy of our in vitro models for filling several important knowledge gaps. The unique ability to model peri-implantation placental development in vitro will enable dissection of the control of MHC-I expression on human trophoblast (TB), particularly testing the hypothesis that the lack of expression of HLA-A and -B by TB is controlled by VTCN1 through largely indirect mechanisms. There are three aims to the proposal: 1) Demonstrate classical MHC class I suppression by VTCN1 in various TB models, including hESC-derived primitive trophoblast, by using VTCN1 knock-down and overexpression; 2) Define the transcriptional pathways and transcriptional and immediate post-transcriptional regulation involved in VTCN1-related alterations in TB MHC-I expression and 3) Identify protein partners of VTCN1 in in vitro cell models of early TB to discover and confirm pathways involved in VTCN1 regulation of MHC class I expression. These studies will have application to the origins of common disorders of abnormal placentation and to the understanding of cancer immunology.

植入植入术的不当局限性发育可能会导致胎儿疾病（早产，intratorine 生长迟钝，先兆子痫）和高妊娠损失率。由于这些事件发生在临床检测妊娠之前，因此在伦理上和逻辑上，对人类占早期发育的研究在伦理上受到限制。拟议的项目使用创新的人类胚胎干细胞（HESC）衍生的模型在体外研究原发性滋养细胞。斑点类似于癌症，因为植入胎儿和一个发育中的癌症都涉及宿主组织入侵，大量的细胞增殖，新的血管化以及立即免疫环境的改变。植入胎儿的位置是唯一显示经典主要组织相容性复合物（MHC）I类（MHC-I）移植抗原，人白细胞抗原（HLA）的生理下调的非癌组织之一。虽然描述了MHC II类调节的阳性调节元件，但对于包括plapeta在内的非癌组织中MHC I类表达的这种调节知之甚少。人类胚胎干细胞（HESC）已用于在临床检测到妊娠之前发生的人类占地发育阶段的新型体外模型。通过使用该模型，已经确定了在早期妊娠早期选择性表达的几种mRNA和蛋白质。在这些研究中，所有人似乎都在入侵中扮演角色。值得注意的是，在我们的早期滋养细胞的模型中，含有T细胞激活抑制剂1（VTCN1）的V-stet结构域之一的siRNA沉默。关于Pleceta特异性HLA-G的表达。 VTCN1被公认为是调节T细胞受体相互作用与MHC分子的B7超家族的成员。据推测，VTCN1新表达会影响癌细胞逃避宿主免疫检测的能力。这些结果突出了我们体外模型填补几个重要知识差距的效率。在体外模拟植入植入术的独特能力将使在人类滋养细胞（TB）上对MHC-I表达的控制进行解剖，尤其是检验了以下假设：TB缺乏HLA-A和-B的表达，通过很大的间接机制由VTCN1控制。该提案的目的有三个：1）通过使用VTCN1敲低和过表达，在包括hESC衍生的原始滋养细胞在内的各种TB模型中展示了VTCN1经典的MHC I类抑制作用； 2）定义参与TB MHC-I中与VTCN1相关改变的转录途径以及转录和直接的转录后调节 表达和3）在早期TB的体外细胞模型中鉴定VTCN1的蛋白质伴侣，以发现和确认参与MHC I类表达的VTCN1调节的途径。这些研究将应用于异常位置的常见疾病的起源和对癌症免疫学的理解。