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Improving the Safety of Genome Editing With Human Kidney Organoids

提高人肾类器官基因组编辑的安全性

基本信息

批准号：
10407081
负责人：
Benjamin Solomon Freedman
金额：
$ 65.28万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-16 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10407081
关键词：
AIDS-Associated Nephropathy Acute Acute Disease Acute Renal Failure with Renal Papillary Necrosis Adverse effects Adverse event Antigen Presentation Architecture Autoimmune Diseases Biological Assay CRISPR/Cas technology Cell Line Cells Chronic Chronic Disease Clinical Trials Computer Models DNA Damage DNA Sequence Distal Early Diagnosis Endothelial Cells Epithelial Cells Event Experimental Models FDA approved Fibrosis Frequencies Genes Genetic Diseases Genome Goals Human Human Genome Immune Impairment In Vitro Inflammation Inflammatory Injury Injury to Kidney Intervention Kidney Kidney Diseases Knowledge Lead Life Lupus Nephritis Malignant Neoplasms Mediating Metabolism Methods Modeling Modification Monitor Mutation Myocardial Infarction Nephritis Nephrons Oncogenic Organ Organoids Outcome Phenotype Physiological Pluripotent Stem Cells Pre-Clinical Model Production Proliferating Renal carcinoma Research Risk Rodent Model Safety Structure Syndrome Therapeutic Time Tissue Model Tissue Sample Tubular formation Work adaptive immune response adverse outcome base carcinogenesis cell type clinical application clinical effect cytokine deep sequencing design genome editing high throughput screening human tissue immunogenic improved in vivo innovation kidney cell podocyte preconditioning predictive modeling replication stress response side effect specific biomarkers therapeutic genome editing therapy development tumorigenesis

项目摘要

IMPROVING THE SAFETY OF GENOME EDITING WITH HUMAN KIDNEY ORGANOIDS PROJECT SUMMARY The goal of this project is to apply genome editors in organoid cultures to establish a predictive model for adverse events in human kidney cell types, including both acute and chronic disorders with life-threatening consequences. Genome editing platforms enable the efficient manipulation of specific DNA sequences in the human genome, and therefore have enormous potential as therapeutics. The kidneys are important target organs, with opportunities for interventions in vivo as well as ex vivo. However, there is a dearth of knowledge about how kidney cells respond to genome editing. Kidneys are known to be susceptible to acute toxic injury, long-term tumorigenesis, as well as chronic immune-mediated responses. A major barrier to predicting these effects is the lack of human experimental models that recapitulate in vivo responses. Rodent models are inherently low-throughput, and have limited ability to predict human safety, while kidney cell lines are too dedifferentiated to accurately model nephrons. To overcome this barrier, we have derived human kidney organoids from pluripotent stem cells as a surrogate for organ structure and function in vitro. Organoids possess many of the key features of kidney nephrons, including diverse cell types in distal-to-proximal arrangements, can express specific phenotypes associated with kidney injury and genetic disease, and are amenable to high throughput screening (HTS). Based on our preliminary work, we hypothesize that gene editing will have deleterious effects on the kidney that are specific, predictable, and can be recapitulated in an organoid model to optimize their design for safe application. This work is of great significance because it will establish a new paradigm for therapy development in human cells, in which multi-dimensional HTS in organoids followed by deep sequencing and detailed analysis identifies the safest and most promising candidates. It is highly innovative because it will bring to light adverse consequences of genome editing of which we are currently unaware, via cutting-edge assays that have never before been applied to organoids. Key findings in organoids will be validated in human kidney tissue samples and human kidneys-on-chips. The proposed research will be pursued as three Aims. The first Aim is to enhance the safety of gene editing for human nephrons by detecting and ameliorating physiological damage to critical cell types in kidney organoids. Aim 2 is to reduce the risk of inadvertent carcinogenesis by profiling oncogenic mutations and transformation events in human kidney organoids subjected to genome editing. Finally, in Aim 3 we propose to identify potential syndromes of editing-associated nephritis by elucidating the immunogenic consequences of CRISPR-Cas9 activity in nephron compartments. Collectively, these three Aims will establish a robust and potentially high throughput framework in which to improve the safety of candidate compounds and increase the number of FDA-approved treatments for kidneys and other organs.

提高人类肾类器官基因组编辑的安全性