Intersection of the mTOR/p70S6K1 signaling and the HIPPO-Yap tissue organizer in neurulation and diabetic embryopathy

mTOR/p70S6K1 信号传导与 HIPPO-Yap 组织组织者在神经形成和糖尿病胚胎病中的交叉点

基本信息

批准号：
10583331
负责人：
Wei-Bin Shen
金额：
$ 65.18万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-21 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10583331
关键词：
70-kDa Ribosomal Protein S6 Kinases Age American Amino Acids Apoptosis Body Size Cell Count Cell Differentiation process Cell Proliferation Cell Size Cell membrane Cell physiology Cells Child Complex Congenital Abnormality Data Defect Development Diabetes Mellitus Embryo Endoplasmic Reticulum Exposure to FRAP1 gene Functional disorder Gene Expression Gene Proteins Growth Homeostasis Human Hyperglycemia Hypertrophy Impairment Induction of Apoptosis Intervention Knockout Mice LATS1 gene Neural Tube Closure Neural Tube Defects Neuroepithelial Cells Neurofibromin 2 Organ Size Pathogenesis Pathway interactions Phosphorylation Phosphorylation Site Phosphotransferases Prenatal care Process Proliferating Proteins Publishing Residual state Scaffolding Protein Signal Induction Signal Transduction Sirolimus Site Structural Congenital Anomalies Testing Tissues Transcription Coactivator Transgenic Mice Transgenic Organisms Translations Woman antagonist cell growth conditional knockout constitutive expression diabetic diabetic embryopathy endoplasmic reticulum stress glycemic control inhibitor insight malformation maternal diabetes neuroepithelium neurogenesis offspring overexpression planar cell polarity prevent reproductive restoration therapeutic target upstream kinase

项目摘要

SUMMARY Pregestational diabetes induces neural tube defects (NTDs). There are 60 million women of reproductive age (18-44 years old) worldwide, and approximately 3 million American women with diabetes. Even under the best prenatal care, women with diabetes are still three- to four-times more likely to have a child with birth defects than women without diabetes. Unraveling the mechanism underlying diabetes-induced NTDs is critical for understanding its pathogenesis and providing potential intervention targets. Yes-associated protein (Yap), a transcriptional co-activator, determines body and organ size and regulates cell apoptosis, proliferation, and differentiation. We found that either conditional KO of Yap or transgenic (Tg) overexpression of constitutively active Yap (CA-Yap) specifically in the neuroepithelium resulted in NTD formation. Our recently published data demonstrate that one of the mTOR (mechanistic target of rapamycin) downstream kinase p70S6K1 is activated by maternal diabetes and its deficiency alleviates diabetic embryopathy. Furthermore, the mTOR inhibitor rapamycin prevents diabetic embryopathy. Both the HIPPO-Yap pathway and the mTOR pathway induce cell and tissue growth. Thus, we hypothesize that maternal diabetes activates mTOR-p70S6K1, which antagonizes Yap by activating the HIPPO kinase Lats1 through phosphorylation, selectively increases translation and disrupts endoplasmic reticulum (ER) homeostasis, leading to NTDs. Lats1 inactivates Yap through phosphorylation that disrupts the planar cell polarity (PCP) leading to failed neurulation. To test our hypothesis, we propose the following aims. Specific Aim 1. determine whether the major mTOR downstream effector p70S6K1 counteracts Yap activity by phosphorylating Yap upstream kinase Lats1 in diabetic embryopathy. We will examine whether maternal diabetes-activated the mTOR effector p70S6K1, which forms a tertiary complex with the scaffold protein Merlin and Lats1, triggers Lats1 phosphorylation in two previously unidentified sites, Thr255 and Thr262, leading to Yap inactivation. Specific Aim 2. To determine whether the HIPPO signaling kinase Lats1 activation leads to neural tube defects in diabetic embryopathy. We will test whether maternal diabetes induces Lats1 phosphorylation at new sites and subsequent the canonical phosphorylation sites which leads to suppression of Yap and NTD formation. Specific Aim 3. To investigate how fine-tuned Yap activity is critical for neurulation and its dysregulation disrupts planar cell polarity in diabetic embryopathy. We will assess that Yap activity is tightly controlled during neurulation and restoring Yap activity can ameliorate diabetic embryopathy. We will further evaluate whether fine-tuned Yap activity maintains PCP gene expression during neurulation.

概括孕育糖尿病会诱导神经管缺陷（NTDS）。有6000万繁殖年龄的妇女（18-44岁）全球，约有300万美国糖尿病女性。即使是最好的产前护理，患有糖尿病的女性患有出生缺陷的可能性仍然高三到四倍没有糖尿病的妇女。阐明糖尿病引起的NTD的机制对于了解其发病机理并提供潜在的干预目标。是相关蛋白（YAP），A 转录共激活因子，确定身体和器官的大小，并调节细胞凋亡，增殖和分化。我们发现，YAP的条件KO或成分性的转基因（TG）过表达专门在神经上皮中的活性YAP（Ca-YAP）导致NTD形成。我们最近发布的数据证明下游激酶P70S6K1的MTOR（机械靶标）之一被激活孕妇糖尿病及其缺乏减轻糖尿病性胚胎。此外，MTOR抑制剂雷帕霉素可预防糖尿病性胚胎。河马途径和mTOR途径都诱导细胞和组织生长。因此，我们假设母体糖尿病激活MTOR-P70S6K1，这通过磷酸化激活HIPPO激酶LATS1来拮抗YAP，有选择地增加翻译和破坏内质网（ER）稳态，导致NTD。 LATS1失活 YAP通过磷酸化破坏了平面细胞极性（PCP），导致神经衰减失败。到检验我们的假设，我们提出以下目标。特定目标1。确定主要MTOR是否下游效应子P70S6K1通过磷酸化YAP上游激酶LATS1来抵消YAP活性在糖尿病性胚胎中。我们将检查产妇糖尿病是否激活MTOR效应子P70S6K1，与脚手架蛋白Merlin和Lats1形成三级复合物，在两个中触发了LATS1磷酸化。以前未识别的地点THR255和THR262导致YAP失活。特定目标2。确定河马信号激酶LATS1激活是否导致糖尿病的神经管缺陷胚胎病。我们将测试产妇糖尿病是否在新地点诱导LATS1磷酸化和随后的规范磷酸化位点，导致YAP和NTD形成抑制。具体的目的3。研究微调YAP活动对于神经肿瘤及其功能失调的关键至关重要糖尿病性胚胎病中的平面细胞极性。我们将评估YAP活动在神经和恢复YAP活性可以改善糖尿病性胚胎病。我们将进一步评估是否微调YAP活性在神经期间保持PCP基因表达。