The Dact/Sestd1 pathway in embryonic malformations

胚胎畸形中的 Dact/Sestd1 通路

基本信息

批准号：
9265875
负责人：
Benjamin N.R. Cheyette
金额：
$ 31.43万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9265875
关键词：
Address Adult Affect Anatomy Animals Architecture Behavior Binding Proteins Biochemical Biochemical Pathway Biological Birth Calcium Calcium Signaling Cardiac Cardiovascular Diseases Cell Shape Cells Child Collection Complex Congenital Abnormality Custom Defect Development Diagnostic Disease Dsh protein Ectoderm Embryo Embryonic Development Endoderm Endosomes Engineering Epithelial Failure Family Family member Funding Gastrointestinal tract structure Genetic Genetically Engineered Mouse Genitourinary system Germ Layers Goals Grant Human Human Development Institutes Integral Membrane Protein Knock-out Knockout Mice Laboratories Link Mammals Membrane Mesenchymal Mesoderm Mission Molecular Mouse Cell Line Movement Mus Mutant Strains Mice Nervous system structure Organ Outcome Pathway interactions Phenocopy Primitive Streaks Protein Family Proteins Proteomics Research Role Scaffolding Protein Signal Pathway Signal Transduction Spectrin Spontaneous abortion Testing Therapeutic Intervention Tissues Transgenic Mice United States Vascular System Vertebrates Vesicle Work Yang biochemical tools blastomere structure cell motility convergent extension design embryo tissue gastrulation improved infant death insight malformation member mutant neonatal surgery neurodevelopment neuropsychiatric disorder novel paralogous gene planar cell polarity prenatal protein degradation protein function protein transport public health priorities public health relevance receptor release of sequestered calcium ion into cytoplasm rho GTP-Binding Proteins therapeutic target tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Human birth defects can be caused by disruption of molecular pathways that regulate cell shape and movements. Cell shape changes and movements are responsible for helping to establish and separate the three primary germ layers at gastrulation, for reification of these germ layers into different tissue types, and for combinin tissues into organs with unique internal architecture and anatomical relationships. Multiple signaling pathways contribute to cell shape and movement, including the Wnt Planar Cell Polarity (PCP) pathway upstream of convergent-extension (CE) movements in vertebrates. In the prior funded period we used an engineered knock-out at the mouse Dapper/Frodo (Dact1) locus to explore how its loss causes a spectrum of developmental defects in mice resembling a major spectrum of birth defects. Through this project we discovered that Dact1 regulates Van Gogh-like (Vangl) transmembrane proteins central to PCP during Epithelial-Mesenchymal Transition (EMT) at the Primitive Streak (PS) during gastrulation. We have now engineered novel knock-out mouse lines for the two remaining Dact family members in mammals (Dact2 and Dact3) as well as for Sestd1, which encodes a Dact-interacting protein identified in my lab. These animals very closely phenocopy Dact1 mutants, strongly supporting the hypothesis that major developmental functions of Sestd1 are allied closely with Dact1. Intriguingly, Sestd1 has previously been shown to help regulate calcium flux through TRPC channel proteins; moreover its closest molecular relative has been implicated in transmembrane protein trafficking. This agrees with evidence suggesting that the molecular pathway emerging from work in my lab acts via localization and/or levels of a subset of transmembrane proteins at the intersection of several important signaling cascades. In the renewed funding period I propose to address these Specific Aims: (1) Do Sestd1 & Dact1 form a genetic pathway with Scribble, Celsr1 & Vangl2 during EMT at the PS; with Dvl2 during CE; and with TRPC4/5 during calcium signaling? (2) Are both the PCP & Calcium pathways biochemically disrupted in posterior embryonic tissues of these mutants? (3) Are membrane localization & levels of pathway components linked to PCP & Calcium signaling in affected embryonic tissues? As in the prior funded period, the research strategy draws on deep mouse genetic, embryonic, cell biologic and biochemical tools custom-designed and assembled in my laboratory expressly to explore this pathway.

描述（由申请人提供）：人类出生缺陷可能是由调节细胞形状和运动的分子途径破坏引起的。细胞形状的变化和运动有助于在原肠胚形成时帮助建立和分离三个主要胚层，将这些胚层具体化为不同的组织类型，并将组织组合成具有独特内部结构和解剖关系的器官。多种信号通路有助于细胞形状和运动，包括脊椎动物中汇聚延伸 (CE) 运动上游的 Wnt 平面细胞极性 (PCP) 通路。在之前的资助期间，我们对小鼠 Dapper/Frodo (Dact1) 基因座进行了工程敲除，以探索其缺失如何导致小鼠出现一系列类似于主要出生缺陷的发育缺陷。通过这个项目，我们发现 Dact1 在原肠胚形成过程中原始条纹 (PS) 的上皮-间质转化 (EMT) 过程中调节 PCP 的核心梵高 (Vangl) 跨膜蛋白。我们现在已经为哺乳动物中剩余的两个 Dact 家族成员（Dact2 和 Dact3）以及 Sestd1 设计了新型敲除小鼠系，Sestd1 编码我实验室鉴定的 Dact 相互作用蛋白。这些动物的表型非常接近 Dact1 突变体，有力地支持了 Sestd1 的主要发育功能与 Dact1 密切相关的假设。有趣的是，Sestd1 先前已被证明有助于调节通过 TRPC 通道蛋白的钙通量；此外，其最接近的分子亲戚与跨膜蛋白质运输有关。这与我实验室工作中出现的分子途径通过在几个重要信号级联交叉点处的跨膜蛋白子集的定位和/或水平起作用的证据相一致。在新的资助期内，我建议解决这些具体目标：(1) Sestd1 和 Dact1 在 PS 的 EMT 期间是否与 Scribble、Celsr1 和 Vangl2 形成遗传途径？ CE 期间使用 Dvl2；以及在钙信号传导过程中与 TRPC4/5 的关系？ (2) 这些突变体的后胚胎组织中的 PCP 和钙途径是否都发生生化破坏？ (3) 膜定位和途径成分水平是否与受影响胚胎组织中的 PCP 和钙信号传导有关？与之前资助期间一样，该研究策略利用了我的实验室专门设计和组装的深层小鼠遗传、胚胎、细胞生物学和生化工具来探索这一途径。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

SEC14 and Spectrin Domains 1 (Sestd1), Dishevelled 2 (Dvl2) and Dapper Antagonist of Catenin-1 (Dact1) co-regulate the Wnt/Planar Cell Polarity (PCP) pathway during mammalian development.

DOI：
10.4161/cib.26834
发表时间：
2013-11-01
期刊：
Communicative & integrative biology
影响因子：
0
作者：
Yang X;Fisher DA;Cheyette BN
通讯作者：
Cheyette BN

Dact1-3 mRNAs exhibit distinct expression domains during tooth development.

DOI：
10.1016/j.gep.2010.02.002
发表时间：
2010-02
期刊：
Gene expression patterns : GEP
影响因子：
0
作者：
Kettunen P;Kivimäe S;Keshari P;Klein OD;Cheyette BN;Luukko K
通讯作者：
Luukko K

All Dact (Dapper/Frodo) scaffold proteins dimerize and exhibit conserved interactions with Vangl, Dvl, and serine/threonine kinases.

DOI：
10.1186/1471-2091-12-33
发表时间：
2011-06-30
期刊：
BMC biochemistry
影响因子：
0
作者：
Kivimäe S;Yang XY;Cheyette BN
通讯作者：
Cheyette BN

DIXDC1 contributes to psychiatric susceptibility by regulating dendritic spine and glutamatergic synapse density via GSK3 and Wnt/β-catenin signaling.

DOI：
10.1038/mp.2016.184
发表时间：
2018-02
期刊：
Molecular psychiatry
影响因子：
11
作者：
Martin PM;Stanley RE;Ross AP;Freitas AE;Moyer CE;Brumback AC;Iafrati J;Stapornwongkul KS;Dominguez S;Kivimäe S;Mulligan KA;Pirooznia M;McCombie WR;Potash JB;Zandi PP;Purcell SM;Sanders SJ;Zuo Y;Sohal VS;Cheyette BNR
通讯作者：
Cheyette BNR