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的破坏引起的。细胞形状的变化和运动负责帮助建立和分离胃肠道的三个主要细菌层，以将这些细菌层重新化为不同的组织类型，并将组合组织组合到具有独特的内部结构和解剖学关系的器官中。多个信号通路有助于细胞形状和运动，包括脊椎动物中收敛扩张（CE）运动上游的Wnt平面细胞极性（PCP）途径。在先前的资助期间，我们使用了小鼠Dapper/Frodo（DACT1）基因座的工程淘汰赛来探索其损失如何导致类似于先天性缺陷的小鼠的一系列发育缺陷。通过该项目，我们发现DACT1在胃胃肠道期间在原始条纹（PS）上调节了上皮 - 间质转变（EMT）期间pCP中心的van Gogh样（VANGL）跨膜蛋白。现在，我们已经为哺乳动物（DACT2和DACT3）以及SESTD1的两个剩余DACT家族成员设计了新型的敲除小鼠系，并编码了我实验室中确定的DACT相互作用蛋白。这些动物非常紧密地表达DACT1突变体，强烈支持以下假设：Sestd1的主要发育功能与DACT1紧密相关。有趣的是，先前已显示SestD1有助于通过TRPC通道蛋白来调节钙通量。此外，其最接近的分子亲戚还与跨膜蛋白运输有关。这与证据表明，我的实验室工作中出现的分子途径通过定位和/或跨膜蛋白的一部分在几个重要信号级联的交汇处。在更新的资金期间，我建议解决这些特定目的：（1）在PS的EMT期间，sestd1＆dact1在EMT期间用涂鸦，celsr1＆vangl2形成遗传途径；在CE期间使用DVL2；在钙信号传导过程中使用TRPC4/5？（2）在这些突变体的后胚胎组织中，PCP和钙途径是否会破坏生化？（3）是否与受影响的胚胎组织中的PCP和钙信号相关的膜定位和途径成分的水平？与先前的时期一样，研究策略借鉴了深层遗传，胚胎，细胞生物学和生化工具，并在我的实验室中定制设计和组装，以明确地探索这一途径。

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