Human pluripotent stem cells

National Institute Of Neurological Disorders And Stroke

Investigators

Linked publications & trials

Abstract

Service and collaboration (disease models) We provided comprehensive training and assistance to ten scientists/trainees from the CC, and NCI, NICHD, NIDCR, NIMH, and NINDS this year: as listed below: ALS (SPT-related) - With Dr. Carsten Bonnemann, NINDS, we are reprogramming fibroblast lines with SPTLC1 and SPTLC2 mutations to iPSCs (SPT-related ALS) and optimizing an in vitro 3D model to obtain mature muscle cells, with the NCATS 3D Tissue Bioprinting Laboratory, to test the effect of gene therapy. ALS4-hiPSC Derivation - With Dr. Christopher Grunseich at NINDS, two fibroblast samples with amyotrophic lateral sclerosis type 4 (ALS4), a rare genetic form of motor neuron disease characterized by late childhood or adolescent onset, are being reprogrammed to iPSCs for motor neuron differentiation to evaluate their phenotypes. Down’s syndrome - Dr. Christopher Bartley, NIMH, plans to study the response of microglia from Down’s Syndrome iPSCs to immunomodulators. We have been generating hiPSC lines from peripheral blood mononuclear cells from these patients as well as controls. GLIS3-mediated differentiation of iPSCs - We previously showed with Dr. Anton Jetton at NIEHS that GLIS3 plays a critical role in pancreatic beta cells, astrocytes, and several other cell types. Protocols are now available to differentiate PSCs into many cell types. We are using differentiation along different cell lineages to obtain further insights into the role of GLIS3 in the regulation of these differentiation pathways and the functions of GLIS3 in these cell types. These strategies should provide a better understanding of the role of GLIS3 in several human diseases, including diabetes, Alzheimer's and cystic renal disease. Muenke Syndrome - With Dr. Fahad Kidwai, NIDCR, we previously reported the craniofacial morphometric analysis and created the world's first human iPSCs from patients with the craniofacial condition, Muenke Syndrome. This project is to perform gene correction using CRISP/Cas9 to make isogeneic lines to comprehend the disease mechanism of craniosynostosis, the major phenotypic finding in these patients. Tumor-induced osteomalacia - Dr. Carlos Ferreira, NICHD, studies tumor-induced osteomalacia (TIO), which is a paraneoplastic syndrome of abnormal phosphorus and vitamin D metabolism, caused most frequently by a translocation of FN1 (fibronectin) and FGFR1 (fibroblast growth factor receptor 1) genes to produce a fusion protein that leads to massive elevation of the phosphaturic hormone, FGF23. The tumor cells that give rise to this condition do not grow in culture. To generate a model for TIO, translocation between the FN1 locus on chromosome 2 and the FGFR1 locus on chromosome 8 has been performed in iPSCs using CRISPR/cas9 technology. Candidate clones are being screened by PCR amplification and sequencing analysis. Research Characterize new pluripotent states and their associated differentiation trajectories - An important property of human pluripotent stem cells (hPSCs) is related to their ground or naive pluripotent state, which may have major impacts on hPSC growth, genetic engineering, disease modeling, and drug discovery. We have derived and comprehensively characterized naive-like hPSCs (NLPs) under various normoxic and hypoxic conditions. Our comparative meta-analysis indicates the existence heterogeneous pluripotent states in diverse NLPs generated from different naive protocols. Interestingly, some NLPs exhibit much lower single cell plating efficiency and commonly lack unique mouse and human NLP marker expression. Evidently, these cells represent an unrecognized minimal naive-like state downstream of formative pluripotency and a trivalent metabolome. Moreover, we revealed a unique metabolome associated with a limited metabolic reprograming capacity in these cells. Our current data provide significant insights into pluripotent state transitions and their associated downstream lineage priming. This research project (Chen et al, Stem Cells, In press).