The Role of MAP 3 kinase 1 in Ocular Surface Morphogenesis

MAP 3 激酶 1 在眼表形态发生中的作用

• 批准号: 7582613
• 负责人: YING XIA
• 金额: $ 37.98万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7582613
• 关键词: Activins; Affect; Architecture; Birth; Cells; Co-Immunoprecipitations; Complex; Cornea; Cultured Cells; DNA Methylation; Defect; Development; Diagnosis; Diagnostic; Disease; Embryo; Environmental Risk Factor; Epigenetic Process; Epithelial; Epithelial Cells; Epithelium; Essential Genes; Etiology; Event; Eye; Eyelid structure; Figs - dietary; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Histone Deacetylase; Human; Intervention; Investigation; JUN gene; Lead; MAP Kinase Kinase Kinase; MAP3K1 gene; Mediating; Mitogen-Activated Protein Kinase 3; Modeling; Modification; Molecular; Molecular Genetics; Molecular Profiling; Molecular Target; Morphogenesis; Mucins; Mus; Mutant Strains Mice; N-terminal; Neoplasm Metastasis; Pathway interactions; Pharmacologic Substance; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Process; Proteins; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Skin; Systems Biology; Testing; Tissues; Tumor Angiogenesis; Two-Dimensional Gel Electrophoresis; Work; Wound Healing; activin B; base; cell motility; chromatin immunoprecipitation; chromatin modification; combinatorial; conjunctiva; histone modification; in utero; in vivo; inhibitor/antagonist; insight; laser capture microdissection; morphogens; ocular surface; postnatal; prognostic; programs; promoter; public health relevance; tool; transcription factor; transmission process

DESCRIPTION (provided by applicant): Tissue morphogenesis is a highly organized and evolutionarily conserved process controlled by the temporal and tissue-specific activation of signal transduction pathways. Vertebrate eyelid morphogenesis undergoes a common process of transient eyelid closure, resulting in closed eyelids that enclose a mucin-enriched conjunctiva sac protecting corneal maturation and development. In mice, eyelid closure takes place at embryonic day 16 (E16) and its re-opening occurs at postnatal day (P)12-14; thus, embryonic eyelid closure defects always result in an eye-open at birth (EOB) phenotype that is easily traceable at birth. Genetic mutant mice displaying EOB therefore constitute unique models to study the molecular factors and complex regulatory mechanisms underlying eyelid morphogenesis. The MAP3 kinase 1 (MAP3K1, also known as MEKK1), an "eyelid closure factor", is responsible for transmission morphogenetic activin B signals to Jun N-terminal kinases (JNKs). JNK in turn phosphorylates transcription factor c-Jun to regulate gene expression events involved in eyelid closure. While activation of the MAP3K1-JNK/Jun pathway by activin B leads to epithelial cell migration, blocking this pathway in the Activin 2B-/-, Map3k1-/- and skin-specific-c-Jun-/- mice, or reduction of the signal transmission efficiency by this pathway in Map3k1Jnk1-/- and Map3k1Jnk1Jnk2 compound mutant mice, all result in defective epithelial cell migration and EOB phenotypes. Despite of the clear contribution of the JNK/Jun pathway, the morphogenetic effects of MAP3K1 depend largely on JNK- independent pathways and crosstalks with other "eyelid closure factors". Based on the findings that MAP3K1 is specifically expressed in the developing eyelid tip epithelial cells, we hypothesize that MAP3K1 is a master gene that coordinates multiple signaling pathways for eyelid epithelial morphogenesis and closure. The proposed studies will use genetic and molecular approaches in combination with systems biology to understand how MAP3K1 orchestrates complex signaling networks in ocular surface morphogenesis. We propose 3 specific aims: (1) to elucidate transcription factor recruitment and epigenetic modifications as mechanisms for Map3k1 promoter activation thereby connecting physiological/environmental factors to eyelid closure; (2) to delineate the downstream pathways responsible for MAP3K1-mediated gene expression; and (3) to understand how MAP3K1 crosstalk with the Smad pathway in coordinating transcription programs for eyelid closure. Human eyelid undergoes strikingly similar morphogenetic processes; however, unlike in mice, eyelid closure and re-opening in humans is entirely accomplished in utero, making it impossible to clinically diagnose eyelid closure defects. Identifying genetic and epigenetic changes associated with eyelid closure defect in mice therefore will offer useful prognostic tool for related congenital ocular diseases in human. Information derived from this work will pave the way for further investigation of genetic and environmental interactions in ocular surface development and identification of the etiology of congenital corneal anomalies. PUBLIC HEALTH RELEVANCE It is a major challenge to understand the mechanism and etiology of human developmental diseases. Results from the proposed work will provide basic understanding and useful prognostic tool for identifying the genetic and epigenetic causes of congenital corneal anomalies in human. Furthermore, our work may lead to mechanistic insights into numerous physiopathological processes in which MAP3K1 participates, such as wound healing, angiogenesis and tumor metastasis.

描述（由申请人提供）：组织形态发生是一种高度组织化和进化保守的过程，由信号转导途径的时间和组织特异性激活控制。脊椎动物眼睑形态发生经历短暂眼睑闭合的常见过程，导致闭合的眼睑包围富含粘蛋白的结膜囊，保护角膜成熟和发育。在小鼠中，眼睑闭合发生在胚胎第16天（E16），其重新打开发生在出生后第12-14天（P）;因此，胚胎眼睑闭合缺陷总是导致出生时睁眼（EOB）表型，这在出生时很容易追踪。因此，显示EOB的遗传突变小鼠构成了研究眼睑形态发生的分子因素和复杂调控机制的独特模型。MAP 3激酶1（MAP 3 K1，也称为MEKK 1），一种“眼睑闭合因子”，负责将形态发生激活素B信号传递至Jun N-末端激酶（JNK）。JNK进而磷酸化转录因子c-Jun以调节参与眼睑闭合的基因表达事件。虽然激活素B激活MAP 3 K1-JNK/Jun途径导致上皮细胞迁移，但在激活素2 B-/-、Map 3 k1-/-和皮肤特异性-c-Jun-/-小鼠中阻断该途径，或在Map 3 k1 Jnk 1-/-和Map 3 k1 Jnk 1 Jnk 2复合突变小鼠中通过该途径降低信号传递效率，均导致上皮细胞迁移缺陷和EOB表型。尽管JNK/Jun通路的贡献明显，但MAP 3 K1的形态发生作用在很大程度上取决于JNK非依赖性通路和与其他“眼睑闭合因子”的串扰。基于MAP 3 K1在发育中的眼睑尖端上皮细胞中特异性表达的发现，我们假设MAP 3 K1是协调眼睑上皮形态发生和闭合的多个信号通路的主基因。拟议的研究将使用遗传和分子方法与系统生物学相结合，以了解MAP 3 K1如何在眼表形态发生中协调复杂的信号网络。我们提出了3个具体目标：（1）阐明转录因子募集和表观遗传修饰作为Map 3 k1启动子激活的机制，从而将生理/环境因素与眼睑闭合联系起来;（2）描绘负责MAP 3 K1介导的基因表达的下游通路;（3）了解MAP 3 K1如何与Smad通路相互作用，协调眼睑闭合的转录程序。人类眼睑经历惊人相似的形态发生过程;然而，与小鼠不同，人类眼睑闭合和重新打开完全在子宫内完成，使得临床上无法诊断眼睑闭合缺陷。因此，识别与小鼠眼睑闭合缺陷相关的遗传和表观遗传变化将为人类相关先天性眼病提供有用的预后工具。从这项工作中获得的信息将为进一步研究眼表发育中的遗传和环境相互作用以及先天性角膜异常的病因学奠定基础。公共卫生相关性了解人类发育性疾病的机制和病因是一项重大挑战。从拟议的工作结果将提供基本的理解和有用的预后工具，以确定遗传和表观遗传的原因，先天性角膜异常的人类。此外，我们的工作可能导致对MAP 3 K1参与的许多生理病理过程的机制见解，如伤口愈合，血管生成和肿瘤转移。