Human pluripotent stem cells

人类多能干细胞

• 批准号: 10016963
• 负责人: Pamela Robey
• 金额: $ 127.32万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 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/10016963
• 关键词: 3-Dimensional; Area; Biological Assay; Biological Process; CRISPR/Cas technology; Cell Culture Techniques; Cell Differentiation process; Cells; Clinic; Clinical; Clone Cells; Collaborations; Communities; Cystic Fibrosis; DNA Sequence Alteration; Data Set; Derivation procedure; Development; Disease; Disease model; Drug Screening; Educational process of instructing; Event; Future; GLIS2 gene; Gene Expression; Generations; Genes; Genomic Instability; Genomics; Goals; Human; In Vitro; Informal Social Control; Kidney; Kidney Diseases; Knock-out; Manuscripts; Membrane; Mentors; Metabolism; Methods; Methylation; Mission; Monitor; Morphogenesis; Mutation; National Institute of Environmental Health Sciences; Nephronophthisis; Netherlands; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Pluripotent Stem Cells; Preclinical Drug Development; Preparation; Process; Proteins; Protocols documentation; Publications; Quality Control; Regenerative Medicine; Regulation; Research Personnel; Role; Sampling; Side; Stem cells; Structure of retinal pigment epithelium; System; TP53 gene; Technology; Tissues; United States National Institutes of Health; Update; Work; X Chromosome; Zinc Fingers; base; cell community; cell growth; cell type; cohort; drug discovery; genome-wide; human embryonic stem cell; human pluripotent stem cell; induced pluripotent stem cell; interest; monolayer; nerve stem cell; personalized predictions; pluripotency; precision medicine; self-renewal; stem cell technology; three dimensional cell culture; tool; web site

During the last fiscal year, the NIH SCCF has made progress in a number of areas as highlighted below. We continued to mentor and teach, both standard and feeder-free, pluripotent stem cell cultures, provided assistance and advice on the generation of human induced pluripotent stem cells (hiPSCs) from collaborators samples, as well as assistance and advice on differentiation strategies as requested. We have also been investigating in-depth the concept of nave pluripotency, which could be important in studying diseases related to the X chromosome. We are currently analyzing large datasets related to gene expression, metabolism and methylation status and a manuscript on this work should be submitted in the coming fiscal year. In terms of bringing pluripotent stem cells to the clinic, we have provided advice on assays and culture of iPSCs related to Dr. Kapil Bhartis (NEI) clinical initiative regarding iPSC derivation and differentiation into retinal pigmented epithelial cells. Traceable markers in human pluripotent stem cell are a valuable tool to study biological processes. To generate various human pluripotent stem cell clones expressing tagged proteins, CRISPR technology has been adapted to introduce GFP or RFP cassettes to the C-termini of endogenous proteins. We are currently generating hPSC lines which express CNS cell type-specific markers for use by the scientific community. We continue to collaborate with Dr. Curtis Harris (NCI), on genomic instability events using a cohort of well-controlled hESC samples. These samples are being examined with various genome-wide technologies and we expect to finish these analyses in the coming fiscal year. These comprehensive analyses will likely provide important information regarding the effect of genomic alterations on human pluripotent stem cell growth under long-term cell culture conditions. Our collaboration with Dr. Ettore Appella (NCI), investigating the role of p53 in the regulation of the self-renewal and differentiation of neural precursors or neural stem cells, is also reaching completion. A manuscript on this work is currently in preparation. In collaboration with Dr. Wei Zheng (NCATS), Dr. Jeffrey Beekman (Regenerative Medicine Center Utrecht, The Netherlands) and Dr. Michael Gottesman (NCI), we considered the use of hPSCs and derivative organoids in drug discovery, in order to define a pathway to implement hPSC-based drug discovery (hPDD). In two publications we discuss specific maladies that employ personalized drug screening (PDS) including cystic fibrosis. We predict that PDS combined with personal genomic analyses will contribute to the development of precision medicine endeavors in the future. Specifically, we dissected representative hPDD systems such as hPSC-based 2D-monolayers, 3D culture, and organoids. We discussed mechanisms of drug discovery and drug repurposing, and the roles of membrane drug transporters in tissue maturation. Following on from a previous collaboration with Dr. Anton Jetten (NIEHS), the role of GLI-similar 2 (GLIS2), which encodes a zinc finger protein, is being investigated. This protein is suggested to have a role in the regulation of kidney morphogenesis with mutations in the gene causing nephronophthisis (NPHP), an autosomal recessive kidney disease. To study the function of GLIS2 protein, the GLIS2 gene has been knocked out using CRISPR technology. In vitro differentiation studies using GLIS2-deficient hESC lines are under way. As always, we update the SCU website with protocols and information as it becomes available to aid other researchers in their studies.

在上一财政年度，NIH SCCF在以下强调的一些领域取得了进展。