ANALYSIS OF DOMINANT MEGACOLON--ANOTHER MODEL FORHIRSCHSPRUNG DISEASE

显性巨结肠分析--先天性巨结肠症的另一种模型

• 批准号: 7968850
• 负责人: William J Pavan
• 金额: $ 100.42万
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7968850
• 关键词: Animals; Back; Cleft Lip; Colon; Complement; Congenital Disorders; Congenital Megacolon; DNA; Defect; Development; Diagnosis; Disease; Embryo; Exhibits; Functional RNA; Ganglia; Gene Targeting; Gene Transfer; Genes; Genetic; Genome; Goals; Human; Hypopigmentation; In Vitro; Investigation; Lead; Maps; Megacolon; Modeling; Mus; Mutagenesis; Mutant Strains Mice; Mutation; Myxoid cyst; Nature; Neural Crest; Pathway interactions; Patients; Peripheral Nervous System; Sampling; Severity of illness; Skin; Stem Cell Development; Stem cells; System; Testing; Transgenic Mice; Variant; Waardenburg syndrome; deafness; gene cloning; in utero; interest; melanocyte; melanoma; mouse model; mutant; overexpression; programs; therapy development; transcription factor; vector

Animals heterozygous for mutants in the SOX10 transcription factor exhibit multiple defects in neural crest development including reduced numbers of melanocytes in the skin, an absence of myenteric ganglion in the colon and can be associated with deafness. Homozgous animals die in utero and there is extensive defects in the entire peripheral nervous system. A human congenital disorder, Hirschsprung disease also exhibits rectocolic aganglionosis and can be associated with hypopigmentation and casued by SOX10 mutations. Thus SOX10 mice, as well as the other neural crest mutant mice, serve as mouse models for this disease. We have found that the SOX10 defects disrupt expression of early neural crest genes, MITF, DCT and EDNRB placing the SOX10 gene early in the neural crest development pathway. We are using additional markers and lineage directed gene transfer to determine the mode of action of SOX10 and its effects on downstream targets. Investigation of the involvement of SOX10 in Hirschsprung disease and other neural crest related disorders will be explored.We have demonstrated that SOX10 directly controls the expression of MITF and DCT. We have shown that the effect on target genes is semindomnant in nature. We have made transgenic mice that overexpress Sxo10 to analyze its effects on neural crest stem cell development. We have also established a system for adding genes back to neural crest stem cells in order to complement genetic defects. We used this system to test hierarchial relationships between SOX10 and its target genes. We have demonstrated that we can use this system to correct SOX10 defects in vitro. We have generated vectors to make this very efficient. We have shown that MITF is not sufficient to complete repalce SOX10 in development. We have also established a whole genome mutagenesis program to identifying SOX10 genetic interaction factors. We have identified 7 heritable loci, mapped five and cloned the mutation in four of the genes. These may become human modifier loci and interesting contributors to neural crest developmental pathways. We are testing humand diseases for mutations in these genes. We have extended the screen to look for earlier embryonic neural crest defects and have mapped four mutants and cloned the gene for three. In addition we have sequenced non-coding DNA from multiple samples of HSCR disease and are looking for correlations of sequence variants disease severity.

Sox10转录因子突变体的动物杂合子在神经rest发育中表现出多种缺陷，包括减少皮肤中的黑素细胞数量，结肠中缺乏肌肠神经节，可能与耳聋有关。纯种动物在子宫内死亡，整个周围神经系统存在广泛的缺陷。人类先天性疾病，Hirschsprung疾病也表现出直肠凝血症，可能与不形成性相关，并由Sox10突变壳。因此，Sox10小鼠以及其他神经rest突变小鼠是该疾病的小鼠模型。我们发现，Sox10缺陷破坏了早期神经rest基因，MITF，DCT和EDNRB的表达，将SOX10基因放置在神经rest发育途径中。我们使用其他标记和谱系定向基因转移来确定SOX10的作用方式及其对下游目标的作用。将研究Sox10参与Hirschsprung疾病和其他神经CREST相关疾病的研究。我们已经证明Sox10直接控制MITF和DCT的表达。我们已经表明，对靶基因的影响本质上是半磁性的。我们已经使过表达SXO10的转基因小鼠分析了其对神经Crest干细胞发育的影响。 我们还建立了一个将基因添加回神经rest干细胞的系统，以补充遗传缺陷。我们使用该系统测试Sox10及其目标基因之间的层次关系。我们已经证明，我们可以使用该系统在体外纠正Sox10缺陷。我们已经生成了向量以使其非常有效。 我们已经表明，MITF不足以完成开发中的Sox10。 我们还建立了一个整个基因组诱变计划，以识别SOX10遗传相互作用因子。我们已经确定了7个可遗传的基因座，绘制了五个基因座，并将突变克隆为四个基因。 这些可能成为人类的修饰基因座，并为神经Crest发育途径而引起有趣的贡献。 我们正在测试这些基因中突变的Humand疾病。 我们已经扩展了屏幕，以寻找早期的胚胎神经rest缺陷，并绘制了四个突变体，并将基因克隆为三个。 此外，我们已经从多个HSCR疾病样本中测序了非编码DNA，并正在寻找序列变体疾病严重程度的相关性。