Regulation of upper lip development by PDGFR Alpha andRac1 signaling

PDGFR Alpha 和 Rac1 信号传导对上唇发育的调节

• 批准号: 8892147
• 负责人: Fenglei He
• 金额: $ 8.56万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-16 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8892147
• 关键词: Affect; Alleles; Automobile Driving; Binding; Birth; Cell Proliferation; Cell physiology; Cells; Cleaved cell; Cleft Lip; Congenital Abnormality; Craniofacial Abnormalities; Data; Development; Disease; Distal; Down-Regulation; Embryonic Development; Event; Exhibits; Face; Future; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Goals; Guanosine Triphosphate; Health; Human; In Vitro; Knockout Mice; Knowledge; Laboratories; Lateral; Ligands; Link; Lip structure; Maxilla; Medial; Mediating; Mediator of activation protein; Methods; Minor; Molecular; Monomeric GTP-Binding Proteins; Morphogenesis; Mus; Mutant Strains Mice; Mutation; Neural Crest Cell; Newborn Infant; Nose; PDGFA gene; PDGFRB gene; Palate; Pathogenesis; Pathway interactions; Pattern; Phenotype; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Platelet-Derived Growth Factor alpha Receptor; Play; Prevention; Process; Proteins; Receptor Protein-Tyrosine Kinases; Regulation; Research; Role; Series; Signal Pathway; Signal Transduction; Testing; Therapeutic; Tissues; Work; cell motility; cleft lip and palate; craniofacial; craniofacial development; design; in vitro Assay; in vivo; innovation; interest; lip morphogenesis; mouse model; mutant; novel; orofacial; orofacial cleft; post-doctoral training; rapid growth; research study; response; transcriptome sequencing

DESCRIPTION (provided by applicant): Cleft lip represents one of most common birth defects and affects approximately 1 in every 700 newborns worldwide. It is caused by disruption of normal upper lip development with environmental or genetic factors. The long term goal of this proposed research is to understand the mechanisms of upper lip development and of orofacial cleft pathogenesis. Mutations of Platelet Derived Growth Factor Receptor ? (PDGFR?) signaling have been tightly linked to cleft lip/palate in humans and mice, suggesting an evolutionarily conserved role in craniofacial development. During embryo development, the upper lip is formed by highly coordinated development of medial nasal process (MNP), lateral nasal process (LNP) and maxillary processes (MxP), all of which are originating from neural crest cells (NCCs). My previous study reveals that PDGFR?? signaling is required to maintain MNP cell proliferation and regulate NCC migration, and indicates a role for small GTPase Rac1 in these processes downstream of PDGFR?. This proposed research is aimed at characterizing the role of Rac1 signaling in mediating PDGFR? regulation on MNP and NCC development. In aim one, I will use a newly generated unbiased Wnt1-Cre2 allele, to ablate Rac1 function specifically in NCCs, and characterize the mutant craniofacial phenotype at histological, cellular and molecular levels. The epistatic effect between Rac1 and PDGFR?? will be tested by inactivating a single allele of each gene in NCCs. I will further rescue the craniofacial phenotype of PDGFR??fl/fl; Wnt1-Cre2 mice by driving expression a constitutively active form of Rac1 in NCCs in vivo. In aim two, I will examine how PDGFR?? signaling regulates Rac1 activity during MNP morphogenesis. I will analyze the activity of immediate downstream signaling pathways in PDGFR?? deficient MNP, and examine of the interaction between the identified signaling pathways and Rac1 activity. Aim three is designed to characterize the expression of PDGFR?? transcriptional targets Cdc42ep3 (CEP3) and its role in craniofacial development. CEP3 will be examined with in vitro assays and gene-targeting method. Results of the proposed works will delineate the signaling cascade by which PDGFR?? regulates NCC and MNP development, at the levels of signaling transduction and transcription. The results of proposed research will provide novel information to understand the fundamental mechanisms of MNP development and upper lip morphogenesis. This study will hold important benefit in treatment of cleft lip, to ultimately reduce the occurrence of this birth defect in humans.

描述（由申请人提供）：唇裂是最常见的出生缺陷之一，全世界每700名新生儿中约有1人受到影响。它是由环境或遗传因素干扰正常上唇发育引起的。本研究的长期目标是了解上唇发育的机制和唇腭裂的发病机制。血小板衍生生长因子受体突变？（PDGFR?）信号与人类和小鼠的唇裂/腭裂密切相关，表明在颅面发育中具有进化保守的作用。在胚胎发育过程中，上唇是由鼻内侧突（MNP）、鼻外侧突（LNP）和上颌突（MxP）高度协调发育形成的，它们都起源于神经嵴细胞（NCCs）。我之前的研究表明PDGFR?？信号传导是维持MNP细胞增殖和调节NCC迁移所必需的，并且表明小GTPase Rac1在PDGFR下游的这些过程中发挥作用。本研究旨在描述Rac1信号在介导PDGFR？对MNP和NCC发展的监管。在目标一中，我将使用一个新产生的无偏倚的Wnt1-Cre2等位基因，在NCCs中特异性地消融Rac1功能，并在组织学，细胞和分子水平上表征突变的颅面表型。Rac1与PDGFR的上位性作用将通过灭活ncc中每个基因的单个等位基因来进行测试。我将进一步挽救颅面表型