Humanized synthetic chromosomes built in yeast for organ xenotransplantation

酵母中内置人源化合成染色体用于器官异种移植

• 批准号: 9305101
• 负责人: David Minh Truong
• 金额: $ 5.92万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-06 至 2018-07-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9305101
• 关键词: Address; Alpha Cell; Animal Model; Artificial Chromosomes; Biochemical; Cells; ChIP-seq; Chromatin; Chromosome Structures; Chromosomes; Chromosomes, Artificial, Mammalian; Cloning; Complex; Computer software; Data; Ensure; Eukaryota; Family suidae; Fellowship; Functional disorder; Gene Expression; Generations; Genes; Genetic; Genome; Goals; Histone H3; Histones; Human; Human Genome; Immune; Immunology; Kinetochores; Knock-out; Lead; Lysine; Maintenance; Methods; Methylation; Modeling; Mutation; Organ; Organ Transplantation; Organism; Pathway interactions; Patients; Pattern; Phenotype; Play; Process; Property; Protein Isoforms; Proteins; Resources; Saccharomyces cerevisiae; Testing; Time; Work; Xenograft procedure; Yeasts; base; cell type; design; gene repression; genetic manipulation; genome-wide; histone methylation; histone modification; homologous recombination; human disease; innovation; insight; pleiotropism; precision genetics; programs; promoter; public health relevance; telomere; tool; transcriptome sequencing; vector; yeast genome

 DESCRIPTION (provided by applicant): The need for human organs far outstrips the supply. One potential transformative solution is to humanize the immune repertoire of pig organs in order to limit patient rejection of xenotransplanted organs. Whole genes and pathways can in principle be put into pig cells using a large artificial chromosome, which the Boeke lab refers to as synthetic Pig Artificial Chromosomes (sPACs). Current versions of these mammalian artificial chromosomes have limitations. The lab will address these issues by completely redesigning the sPACs from the `bottom up' in the yeast Saccharomyces cerevisiae. These yeast-based sPACs could be readily reprogrammed with patient data and quickly uploaded into pig cells. In order to achieve this `plug and play' capability, the primary goal of this postdoctoral proposal is to humanize the chromatin of S. cerevisiae, thus ensuring the generation of mammalian-like chromosome structure. The proposal will focus on three specific aims: 1) humanize the histones of yeast; 2) build in H3K27 methylation; and 3) build in H3K9 methylation. In addition, the proposal will permit the study of the de novo establishment of mammalian chromatin in a cell. This will lead to fundamental insight into methylation establishment, maintenance, and mechanisms of perturbation, information relevant to human gene expression. This proposal will achieve two innovative and complementary goals by building human chromatin into yeast: study the mechanisms of chromatin establishment in a cellular 'blank slate', and generate a cell capable of making pig artificial chromosomes designed to facilitate organ xenotransplantation. The long-term goal is to provide an unlimited method of generating humanized pig organs for organ transplantation, thus advancing the treatment of human disease.

 描述（由申请人提供）：人体器官的需求远远超过供应。一种潜在的变革性解决方案是将猪器官的免疫系统人性化，以限制患者对异种移植器官的排斥。原则上可以使用大型人工染色体将整个基因和通路放入猪细胞中，Boeke 实验室将其称为合成猪人工染色体（sPAC）。这些哺乳动物人工染色体的当前版本有局限性。该实验室将通过在酿酒酵母中“自下而上”彻底重新设计 sPAC 来解决这些问题。这些基于酵母的 sPAC 可以很容易地用患者数据重新编程，并快速上传到猪细胞中。为了实现这种“即插即用”的能力，该博士后提案的主要目标是使酿酒酵母的染色质人源化，从而确保生成类似哺乳动物的染色体结构。该提案将重点关注三个具体目标：1）将酵母组蛋白人性化； 2) 内置H3K27甲基化； 3) 构建 H3K9 甲基化。此外，该提案还将允许研究哺乳动物染色质在细胞中的从头建立。这将带来对甲基化建立、维持和扰动机制以及与人类基因表达相关的信息的基本了解。该提案将通过将人类染色质构建到酵母中来实现两个创新和互补的目标：研究细胞“空白石板”中染色质建立的机制，并生成能够制造猪人工染色体的细胞，旨在促进器官异种移植。长期目标是为器官移植提供无限的人源化猪器官生成方法，从而推进人类疾病的治疗。