Ahead of the Curve: Mechanisms of Left-Right Asymmetric Stomach Morphogenesis

引领潮流：左右不对称胃形态发生的机制

• 批准号: 10397529
• 负责人: Nanette M Nascone-Yoder
• 金额: $ 30.23万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-17 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10397529
• 关键词: Address; Adopted; Amphibia; Anatomy; Animal Model; Behavior; Bilateral; Biological; Biological Assay; Candidate Disease Gene; Cell physiology; Cells; Clinical; Complement; Complex; Congenital Abnormality; Cues; Data; Data Set; Defect; Development; Digestive Physiology; Dissection; Economic Burden; Embryo; Etiology; Event; Family; Gene Expression; Genes; Genetic; Goals; Handedness; Individual; Infant Mortality; Knowledge; Left; Life; Logic; Mammals; Manuals; Modeling; Molecular; Molecular Profiling; Morphogenesis; Nodal; Organ; Organogenesis; Pathway interactions; Pattern; Phase; Phenotype; Positioning Attribute; Process; Property; Radial; Rana; Regulator Genes; Research; Shapes; Side; Situs Inversus; Stomach; Structural Congenital Anomalies; Testing; Thinness; Tissues; Tube; United States; Vertebrates; Xenopus; body system; driving force; gain of function; gene discovery; gene regulatory network; innovation; insight; malformation; novel; programs; spatiotemporal; transcriptome; transcriptome sequencing; validation studies

Project Summary Birth defects are a leading cause of infant mortality, yet in most cases, their etiology is unknown. Some of the most common and complex malformations are found in families with abnormal left-right (LR) asymmetry, suggesting that many congenital defects may result from perturbed laterality. The initial embryonic events that determine the LR body axis, including the early breaking of bilateral symmetry, and subsequent left-sided expression of determinants such as nodal and Pitx2, are now well understood. However, the genetic and morphogenetic events involved in the final phases of LR development, at the organ level, remain largely unknown. The long term goal is to ascertain the mechanisms of LR asymmetric organogenesis. The objective in this application is to identify the molecular and cellular processes that generate LR asymmetry (curvature) within an individual organ, the stomach. Preliminary analyses identified LR asymmetries in radial cell rearrangements in the developing stomach as the driving force for its curvature. To identify the proximate effectors of this novel asymmetric morphogenetic program, a new model organism (Lepidobatrachus laevis) was employed. The extra-large embryos of this species facilitated a gene-discovery approach that would be intractable in most models: transcriptome profiling (RNASeq) of tissues dissected from left vs. right halves of the embryonic stomach. Pilot datasets include genes with LR asymmetric expression patterns and functions during stomach curvature. The central hypothesis is that stomach curvature is determined by distinct left and right regulatory networks which differentially modulate the cellular events controlling radial cell rearrangement. The unique experimental amenability of frog embryos will be used to test this hypothesis via three specific aims: 1) Generate molecular signatures of normal and abnormal stomach curvature. Comprehensive spatiotemporal profiles of LR stomach genes will be generated and compared in the context of both normal LR asymmetry and experimentally-induced LR axis defects. 2) Determine the cellular function of stomach- specific LR genes. Select genes will be tested in loss- and gain-of-function assays to determine their influence on radial cell rearrangement in the developing stomach. 3) Determine the regulatory hierarchy that controls stomach curvature. Experimental perturbations combined with spatiotemporal profiling will reveal core gene regulatory interactions governing asymmetric morphogenesis. The overall approach is innovative because it takes advantage of distinctive attributes of a unique species to address one of the key unanswered questions in LR development: what are the proximate mechanisms by which developing organs become LR asymmetric entities? The proposed research is significant because it will immediately advance our understanding of normal laterality and laterality-related birth defects by defining the genes, morphogenetic processes and regulatory logic that govern the emergence of LR asymmetry at the organ level.

项目摘要 出生缺陷是婴儿死亡率的主要原因，但在大多数情况下，其病因尚不清楚。一些 在最常见和最复杂的畸形中，有异常左右（LR）不对称性的家族， 表明许多先天性缺陷可能是由于横向扰动而导致的。最初的胚胎事件 确定LR身体轴，包括双侧对称性的早期断裂，然后左侧 现在已经充分理解了决定因素（例如节点和pitx2）的表达。但是，遗传和 在器官水平上，LR发育最终阶段涉及的形态发生事件在很大程度上保持不变 未知。长期目标是确定LR非对称器官发生的机制。目标 在此应用中，是确定产生LR不对称性（曲率）的分子和细胞过程 在单个器官中，胃。初步分析确定了径向细胞中的LR不对称性 发育中的胃中的重排作为其曲率的驱动力。识别近距离 这个新型不对称形态学程序的效应因素，一种新的模型生物（鳞翅目Laevis） 被雇用。该物种的超大胚胎促进了一种基因发现方法 在大多数模型中棘手：从左侧与右半部分解剖的组织的转录组分析（RNASEQ） 胚胎胃。试验数据集包括具有LR不对称表达模式和功能的基因 在胃弯曲期​​间。中心假设是胃弯曲是由不同的左左和 右监管网络会差异地调节控制径向细胞重排的细胞事件。 青蛙胚胎的独特实验性舒适性将用于通过三个特定的特定 目的：1）产生正常和异常胃曲率的分子特征。综合的 LR胃基因的时空特征将被产生，并在两种正常情况下进行比较 LR不对称和实验诱导的LR轴缺陷。 2）确定胃的细胞功能 特定的LR基因。选定的基因将在损失和功能获得分析中进行测试，以确定其 对发育中胃中径向细胞重排的影响。 3）确定监管层次结构 控制胃弯曲。实验性扰动与时空分析相结合将 揭示了控制不对称形态发生的核心基因调节相互作用。总体方法是 创新性是因为它利用独特物种的独特属性来解决关键之一 LR开发中未解决的问题：开发器官的近端机制是什么 成为LR非对称实体？拟议的研究很重要，因为它将立即前进 我们通过定义基因的形态发生对正常横向性和与侧向相关的出生缺陷的理解 控制LR不对称性在器官水平的过程和调节逻辑。

项目成果

Morphoelastic models discriminate between different mechanisms of left-right asymmetric stomach morphogenesis.

形态弹性模型区分左右不对称胃形态发生的不同机制。

• DOI: 10.1016/j.cdev.2024.203902
• 发表时间: 2024
• 期刊: Cells & development
• 影响因子: 3.9
• 作者: Nikas,ArielN;Curcio,EvanJ;Nascone-Yoder,Nanette;Lubkin,SharonR
• 通讯作者: Lubkin,SharonR

The twists and turns of left-right asymmetric gut morphogenesis.

左右不对称肠道形态发生的曲折。

• DOI: 10.1242/dev.187583
• 发表时间: 2020
• 期刊: Development (Cambridge, England)
• 作者: Grzymkowski,Julia;Wyatt,Brent;Nascone-Yoder,Nanette
• 通讯作者: Nascone-Yoder,Nanette

Single-minded 2 is required for left-right asymmetric stomach morphogenesis.

左右不对称的胃形态发生需要“一心一意”2。

• DOI: 10.1242/dev.199265
• 发表时间: 2021
• 期刊: Development (Cambridge, England)
• 作者: Wyatt,BrentH;Amin,NiravM;Bagley,Kristen;Wcisel,Dustin;Dush,MichaelK;Yoder,JeffreyA;Nascone-Yoder,NanetteM
• 通讯作者: Nascone-Yoder,NanetteM

Rare variants in CAPN2 increase risk for isolated hypoplastic left heart syndrome.

• DOI: 10.1016/j.xhgg.2023.100232
• 发表时间: 2023-10-12
• 期刊: HUMAN GENETICS AND GENOMICS ADVANCES
• 作者: Blue, Elizabeth E.;White, Janson J.;Dush, Michael K.;Gordon, William W.;Wyatt, Brent H.;White, Peter;Marvin, Colby T.;Helle, Emmi;Ojala, Tiina;Priest, James R.;Jenkins, Mary M.;Almli, Lynn M.;Reefhuis, Jennita;Pangilinan, Faith;Brody, Lawrence C.;McBride, Kim L.;Garg, Vidu;Shaw, Gary M.;Romitti, Paul A.;Nembhard, Wendy N.;Browne, Marilyn L.;Werler, Martha M.;Kay, Denise M.;Mital, Seema;Chong, Jessica X.;Nascone-Yoder, Nanette M.;Bamshad, Michael J.
• 通讯作者: Bamshad, Michael J.