Microphysiological Human Tissue Systems for Monitoring of Genome Editing Outcomes

用于监测基因组编辑结果的微生理人体组织系统

• 批准号: 10477016
• 负责人: Nenad Bursac
• 金额: $ 66.82万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10477016
• 关键词: Address; Affect; Animal Model; Architecture; Bacteria; Biology; CRISPR/Cas technology; Cell Line; Cell Survival; Cells; Cellular Immunity; Cellular Metabolic Process; Clinical; Communicable Diseases; DNA Sequence Alteration; Development; Disease; Disease model; Duchenne muscular dystrophy; Ensure; Evaluation; Genes; Genetic Diseases; Genome; Genome Stability; Genome engineering; Genomics; Goals; Human; Human Biology; Human Engineering; Human Genome; Humoral Immunities; Immune; Immune response; Immune system; Immunity; Injury; Methods; Modification; Monitor; Mus; Muscle; Muscle Development; Muscle satellite cell; Myocardium; Natural regeneration; Non-Viral Vector; Nucleic Acids; Outcome; Physiological; Physiology; Pluripotent Stem Cells; Prevalence; Proteins; Regenerative response; Research Personnel; Role; Severities; Site; Site-Directed Mutagenesis; Skeletal Muscle; System; Technology; Therapeutic; Tissue Engineering; Tissues; Translating; Translations; Treatment Efficacy; United States National Institutes of Health; Viral; adaptive immune response; adverse outcome; delivery vehicle; design; drug discovery; experimental study; genome analysis; genome editing; genome integrity; genome-wide; human disease; human tissue; insight; microphysiology system; muscle form; new technology; novel strategies; off-target site; response; somatic cell gene editing; stem cells; therapeutic genome editing; therapy outcome; tissue injury; tissue regeneration; tissue stem cells; tool; vector

Abstract: Genome editing technologies have significant potential to treat a variety of devastating human diseases and disorders. However, there are a number of challenges that genome editing therapies must overcome to reach their full promise. Specifically, there are many possible adverse consequences that are unique to genome editing tools, such as genome integrity, immune responses, and loss of therapeutic efficacy due to cell turnover, for which there are currently are no optimal systems for rigorous assessment. Moreover, these consequences are unique to human physiology, genome sequence, and immune systems, and therefore typical animal models are not completely informative. To address this unmet need, we have assembled a team of collaborative investigators that have developed advanced genome editing strategies and methods for engineering human microphysiological tissue systems that recapitulate human physiology and function, with an emphasis on skeletal and cardiac muscle. We will combine these technologies in this proposal to systemically evaluate tissue physiology, genomic alterations, tissue regeneration, and immune response in response to various genome editing strategies and delivery methods. Specifically, this will include comprehensive and unbiased mapping of unintended modifications to human genome sequences, including at on-target and off-target sites. We will also determine the role of resident tissue stem cells, cell turnover, and tissue injury and regeneration in the stability of genome editing. Finally, we can incorporate immune cells into these microphysiological tissues to understand the consequences of immunity to bacteria-derived genome editing components. Collectively, this proposal will develop a platform to systematically address the most significant challenges to realizing the transformative potential of genome editing therapies in human tissues.

抽象的： 基因组编辑技术具有治疗多种毁灭性人类疾病的巨大潜力 失调。然而，基因组编辑疗法必须克服许多挑战才能实现 他们的全部承诺。具体来说，基因组编辑特有的许多可能的不利后果 工具，例如基因组完整性、免疫反应以及由于细胞更新而导致的治疗功效丧失， 目前尚无最佳系统进行严格评估。而且，这些后果是 人类生理学、基因组序列和免疫系统所独有，因此典型的动物模型是 信息不完全。为了解决这一未满足的需求，我们组建了一个协作团队 研究人员开发了先进的基因组编辑策略和方法来工程人类 概括人体生理学和功能的微观生理组织系统，重点是骨骼 和心肌。我们将在本提案中结合这些技术来系统地评估组织 生理学、基因组改变、组织再生和对各种基因组的免疫反应 编辑策略和交付方法。具体来说，这将包括全面、公正地绘制 对人类基因组序列的意外修改，包括在目标位点和脱靶位点的修改。我们也会 确定常驻组织干细胞、细胞更新以及组织损伤和再生在稳定性中的作用 基因组编辑。最后，我们可以将免疫细胞整合到这些微生理组织中来了解 对细菌衍生的基因组编辑成分产生免疫力的后果。总的来说，该提案将 开发一个平台来系统地解决实现变革的最重大挑战 基因组编辑疗法在人体组织中的潜力。

项目成果

A humanized mouse model for adeno-associated viral gene therapy.

用于腺相关病毒基因治疗的人源化小鼠模型。

• 发表时间: 2024-03-04
• 影响因子: 16.6
• 作者: Barzi, Mercedes;Chen, Tong;Gonzalez, Trevor J;Pankowicz, Francis P;Oh, Seh Hoon;Streff, Helen L;Rosales, Alan;Ma, Yunhan;Collias, Sabrina;Woodfield, Sarah E;Diehl, Anna Mae;Vasudevan, Sanjeev A;Galvan, Thao N;Goss, John;Gersbach, Charles A
• 通讯作者: Gersbach, Charles A