TILLING wheat for reduced celiac disease causing proteins

耕种小麦以减少引起乳糜泻的蛋白质

• 批准号: 8143121
• 负责人: Charles Paul Moehs
• 金额: $ 6.64万
• 依托单位: ARCADIA BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8143121
• 关键词: Address; Adherence; Affect; Alleles; Amino Acid Sequence; Autoimmune Process; Barley; Binding; Boxing; Bread; Breeding; Candidate Disease Gene; Celiac Disease; Cereals; Chromosomes; Collaborations; CpG Islands; DNA; DNA Binding; Development; Diet; Digestion; Digestive System Disorders; Disease; Epitopes; Exclusion; Fingers; Food; Gastroenterology; Gene Expression; Gene Family; Gene Silencing; Genes; Genetic Transcription; Genetic Transformation; Genetically Modified Organisms; Genome; Gliadin; Gluten; Homologous Gene; Human; Immune; Individual; Ingestion; Intestines; Lead; Lesion; Lysine; Methods; Molecular Weight; Mutation; Nonsense Mutation; Nutritional; Outcome; Outcomes Research; Patients; Peptide Sequence Determination; Peptides; Phase; Phenotype; Plants; Play; Prevalence; Property; Proteins; Reaction; Research; Research Support; Resistance; Resources; Role; Science; Scientist; Seeds; Self-Fertilizations; Small Business Technology Transfer Research; Starch; Techniques; Testing; Transgenic Organisms; Translations; United States National Institutes of Health; Universities; Variant; Washington; Wheat; Work; commercialization; demethylation; expression cloning; glutenin; improved; innovation; mutant; novel; plant prolamin protein; prevent; promoter; public health relevance; response; transcription factor

DESCRIPTION (provided by applicant): Celiac disease is an autoimmune digestive disorder in which intestinal lesions develop in genetically susceptible individuals in response to the ingestion of specific cereal seed storage proteins. The only treatment is strict adherence to a diet free from the offending grains. Therefore, considerable research has been devoted to identifying the specific amino acid sequences in wheat seed storage proteins responsible for eliciting the immune reaction in celiac patients, and this research has motivated the hope that strains of wheat and other grains could be developed in which these proteins have been reduced or eliminated. During the course of NIH STTR phase I research conducted in collaboration between Dr. Diter von Wettstein of Washington State University and Arcadia Biosciences, we have cloned and begun a systematic analysis of the composition of the previously uncharacterized ? gliadin gene family located on homoeologous chromosome 6D of hexaploid wheat. Among this gliadin gene family there exist one or more genes for the gliadin subtype immunodominant in celiac disease containing a 33-mer peptide highly resistant to digestion (Molberg et al., Gastroenterology 128:393-401,2005; Shan et al., Science 297:2275-2279,2002). Identifying the gene(s) encoding this gliadin subtype is a prerequisite for using TILLING to attempt to eliminate it from the wheat grain. TILLING (Targeting Induced Local Lesions in Genomes) is a novel, non-GMO (genetically modified organism) technique invented by an Arcadia Biosciences scientist that is a target-selected variation of mutation breeding. During STTR phase I research we have also determined the importance of a DNA 5- methylcytosine deglycosylase for gluten gene expression in wheat and barley and are targeting the wheat homologues for inactivation via TILLING and transgenic means. Finally, we have developed a large number of novel wheat lines containing missense and nonsense mutations in candidate transcription factors known to be important for the accumulation of seed storage proteins toxic to celiac patients. STTR phase II research will continue progress towards the development of wheat acceptable for celiac patients by: 1. Using TILLING and transgenic approaches to develop wheat plants down-regulated for endosperm DNA 5- methylcytosine deglycosylation activity and characterizing their seed storage composition. 2. Making crosses between plants harboring priority missense and nonsense alleles in the candidate transcription factors and characterizing the phenotypes of the progeny. The outcome of this NIH STTR Phase II research will be mutant and transgenic wheat lines whose creation are a necessary prerequisite for the commercialization of wheat cultivars acceptable for sufferers of celiac disease. PUBLIC HEALTH RELEVANCE: The proposed research takes an innovative approach to ameliorating the cause of celiac disease by developing wheat varieties that lack the major gluten components that elicit the disease. Celiac disease is the most common food-sensitive enteropathological condition in humans with a prevalence of 0.5-1%, and currently the only treatment for patients is the complete exclusion of cereal proteins from the diet. Developing wheat that is acceptable for celiac patients thus addresses an urgent, unmet need.

描述（由申请人提供）：乳糜泻是一种自身免疫性消化系统疾病，其中肠道病变在遗传易感个体中发生，以响应特定谷物种子贮藏蛋白的摄入。唯一的治疗方法是严格遵守不含有害谷物的饮食。因此，大量的研究致力于鉴定小麦种子贮藏蛋白中负责引发乳糜泻患者免疫反应的特定氨基酸序列，并且这项研究激发了希望，可以开发出小麦和其他谷物的菌株，其中这些蛋白质已经减少或消除。在美国国立卫生研究院STTR第一阶段的研究过程中，华盛顿州立大学和阿卡迪亚生物科学公司的Diter von Wettstein博士合作进行，我们已经克隆并开始系统分析以前未表征的？醇溶蛋白基因家族位于六倍体小麦同源染色体6D上。在该麦醇溶蛋白基因家族中，存在一个或多个在乳糜泻中免疫显性的麦醇溶蛋白亚型的基因，其含有高度耐消化的33-mer肽（Molberg et al.，Gastroenterology 128：393- 401，2005; Shan等人，Science 297：2275- 2279，2002）。鉴定编码这种麦醇溶蛋白亚型的基因是使用TILLING试图从小麦籽粒中消除它的先决条件。TILLING（靶向诱导基因组局部病变）是一种新型的非转基因（转基因生物）技术，由Arcadia Biosciences科学家发明，是一种目标选择的突变育种变异。在STTR I期研究期间，我们还确定了DNA 5-甲基胞嘧啶去糖基化酶对小麦和大麦中谷蛋白基因表达的重要性，并通过TILLING和转基因手段靶向小麦同源物进行失活。最后，我们已经开发了大量的新的小麦品系含有错义和无义突变的候选转录因子已知是重要的积累的种子储存蛋白有毒的乳糜泻患者。STTR II期研究将继续朝着开发乳糜泻患者可接受的小麦的方向发展：1。使用TILLING和转基因方法开发胚乳DNA 5-甲基胞嘧啶去糖基化活性下调的小麦植株，并表征其种子贮藏组成。2.在候选转录因子中具有优先错义和无义等位基因的植物之间进行杂交，并表征后代的表型。这项NIH STTR第二阶段研究的结果将是突变和转基因小麦品系，其创造是乳糜泻患者可接受的小麦品种商业化的必要先决条件。公共卫生相关性：这项拟议中的研究采取了一种创新的方法，通过开发缺乏引起这种疾病的主要面筋成分的小麦品种来改善乳糜泻的病因。乳糜泻是人类中最常见的食物敏感性肠道病理状况，患病率为0.5- 1%，目前对患者的唯一治疗方法是从饮食中完全排除谷物蛋白。因此，开发乳糜泻患者可接受的小麦解决了一个紧迫的，未满足的需求。