A5001173505- Pluripotent Stem Cells Research
- CRISPR and Genetic Engineering
- Lysosomal Storage Disorders Research
- Virus-based gene therapy research
- Neuroinflammation and Neurodegeneration Mechanisms
- Renal and related cancers
- Retinal Development and Disorders
- Animal Genetics and Reproduction
- Retinal Diseases and Treatments
- RNA modifications and cancer
- Zebrafish Biomedical Research Applications
- Epigenetics and DNA Methylation
- Genetics and Neurodevelopmental Disorders
- Pancreatic function and diabetes
- RNA Interference and Gene Delivery
- Neuroscience and Neural Engineering
- CAR-T cell therapy research
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Neurogenesis and neuroplasticity mechanisms
- Cellular transport and secretion
- Endoplasmic Reticulum Stress and Disease
- Child Nutrition and Feeding Issues
- Pediatric Hepatobiliary Diseases and Treatments
- 3D Printing in Biomedical Research
- Developmental Biology and Gene Regulation
National Heart Lung and Blood Institute
2016-2025
National Institutes of Health
2016-2025
Xiangya Hospital Central South University
2022
Central South University
2022
Johns Hopkins Medicine
2008-2019
Johns Hopkins University
2008-2019
Center for Neuroscience and Regenerative Medicine
2013-2019
Center for Molecular Medicine and Immunology
2016
National Institute of Arthritis and Musculoskeletal and Skin Diseases
2013-2015
Stem Cell Institute
2011-2012
Abstract We report here that butyrate, a naturally occurring fatty acid commonly used as nutritional supplement and differentiation agent, greatly enhances the efficiency of induced pluripotent stem (iPS) cell derivation from human adult or fetal fibroblasts. After transient butyrate treatment, iPS is enhanced by 15- to 51-fold using either retroviral piggyBac transposon vectors expressing 4 5 reprogramming genes. Butyrate stimulation more remarkable (>100- 200-fold) on in absence...
Abstract It was reported recently that human fibroblasts can be reprogrammed into a pluripotent state resembles of embryonic stem (hES) cells. This achieved by ectopic expression four genes followed culture on mouse fibroblast (MEF) feeders under condition favoring hES cell growth. However, the efficiency generating induced (iPS) cells is low, especially for postnatal fibroblasts. We started supplementing with an additional gene or bioactive molecules to increase iPS from adult as well fetal...
Human induced pluripotent stem cell (iPSC) lines are a powerful tool for studying development and disease, but the considerable phenotypic variation between makes it challenging to replicate key findings integrate data across research groups. To address this issue, we sub-cloned candidate human iPSC deeply characterized their genetic properties using whole genome sequencing, genomic stability upon CRISPR-Cas9-based gene editing, including differentiation commonly used types. These studies...
The derivation of three-dimensional (3D) stratified neural retina from pluripotent stem cells has permitted investigations human photoreceptors. We have generated a H9 embryonic cell subclone that carries green fluorescent protein (GFP) reporter under the control promoter cone-rod homeobox (CRX), an established marker postmitotic photoreceptor precursors. CRXp-GFP replicates endogenous CRX expression in vitro when is induced to form self-organizing 3D retina-like tissue. At day 37, CRX+...
Targeted genome engineering to robustly express transgenes is an essential methodology for stem cell-based research and therapy. Although designer nucleases have been used drastically enhance gene editing efficiency, targeted addition stable expression of date limited at single gene/locus mostly PPP1R12C/AAVS1 in human cells. Here we constructed transcription activator-like effector (TALENs) targeting the safe-harbor like CLYBL mediate reporter integration 38%–58% both AAVS1-TALENs...
The polycomb repressive complex 2 (PRC2) methylates lysine 27 of histone H3 (H3K27) through its catalytic subunit Ezh2. PRC2-mediated di- and tri-methylation (H3K27me2/H3K27me3) have been interchangeably associated with gene repression. However, it remains unclear whether these two degrees H3K27 methylation different functions. In this study, we generated isogenic mouse embryonic stem cells (ESCs) a modified H3K27me2/H3K27me3 ratio. Our findings document dynamic developmental control in the...
Abstract Human induced pluripotent stem (hiPS) cell lines with tissue-specific or ubiquitous reporter genes are extremely useful for optimizing in vitro differentiation conditions as well monitoring transplanted cells vivo. The adeno-associated virus integration site 1 (AAVS1) locus has been used a “safe harbor” inserting transgenes because of its open chromatin structure, which permits transgene expression without insertional mutagenesis. However, it is not clear whether targeted at the...
There are five genetic forms of chronic granulomatous disease (CGD), resulting from mutations in any subunits phagocyte oxidase, an enzyme complex neutrophils, monocytes, and macrophages that produces microbicidal reactive oxygen species. We generated induced pluripotent stem cells (iPSCs) peripheral blood CD34(+) hematopoietic patients with each CGD genotypes. used zinc finger nuclease (ZFN) targeting the AAVS1 safe harbor site together genotype-specific minigene plasmids flanking sequence...
Significance Doxorubicin is a DNA-damaging agent that highly effective against various types of cancers, but subset treated patients develop heart failure for unclear genetic reasons. In the current study using p53 mouse models, low-dose doxorubicin as administered in clinics surprisingly revealed absence increases susceptibility to cardiotoxicity while mutant retains mitochondrial regulation protective. Notably, promoting biogenesis with simple vitamin supplement ameliorated cardiotoxicity....
Cardiomyocytes can be differentiated from human pluripotent stem cells (hPSCs) in defined conditions, but efficient and consistent cardiomyocyte differentiation often requires expensive reagents such as B27 supplement or recombinant albumin. Using a chemically albumin-free (E8 basal) medium, we identified heparin novel factor that significantly promotes efficiency, developed an method to differentiate hPSCs into cardiomyocytes. The treatment with helped consistently reach at least 80% purity...
Efficient and homogeneous in vitro generation of peripheral sensory neurons may provide a framework for novel drug screening platforms disease models touch pain. We discover that, by overexpressing NGN2 BRN3A, human pluripotent stem cells can be transcriptionally programmed to differentiate into surprisingly uniform culture cold- mechano-sensing neurons. Although such neuronal subtype is not found mice, we identify molecular evidence its existence ganglia. Combining BRN3A programming with...
Embryos devoid of autonomic innervation suffer sudden cardiac death. However, whether neurons have a role in heart development is poorly understood. To investigate if sympathetic impact cardiomyocyte maturation, we co-cultured phenotypically immature cardiomyocytes derived from human induced pluripotent stem cells with mouse ganglion neurons. We found that 1) multiple structure and ion channel genes related to maturation were up-regulated when neurons; 2) sarcomere organization connexin-43...
TDP-43 mislocalization and pathology occurs across a range of neurodegenerative diseases, but the pathways that modulate in neurons are not well understood. We generated Halo-TDP-43 knock-in iPSC line performed genome-wide CRISPR interference FACS-based screen to identify modifiers levels neurons. A meta-analysis our publicly available screens identified both specific hits present multiple screens, latter likely responsible for generic protein level maintenance. BORC, complex required...
TDP-43 mislocalization and pathology occurs across a range of neurodegenerative diseases, but the pathways that modulate in neurons are not well understood. We generated Halo-TDP-43 knock-in iPSC line performed genome-wide CRISPR interference FACS-based screen to identify modifiers levels neurons. A meta-analysis our publicly available screens identified both specific hits present multiple screens, latter likely responsible for generic protein level maintenance. BORC, complex required...
Nonsense mediated mRNA decay (NMD) is an RNA surveillance mechanism that controls stability and ensures the speedy degradation of erroneous unnecessary transcripts. This depends on several core factors in exon junction complex (EJC), eIF4A3, RBM8a, Magoh, BTZ, as well peripheral to distinguish premature stop codons (PTCs) from normal Recently, emerging evidence has indicated NMD are associated with neurodevelopmental disorders such autism spectrum disorder (ASD) intellectual disability (ID)....
X-linked chronic granulomatous disease (X-CGD) is an immune deficiency resulting from defective production of microbicidal reactive oxygen species (ROS) by phagocytes. Causative mutations occur throughout the CYBB gene, in absent or gp91phox protein expression. To correct exon 5 while retaining normal gene regulation, we utilized TALEN Cas9 for replacement induced pluripotent stem cells (iPSCs) patients, which restored expression and ROS iPSC-derived granulocytes. Alternate approaches...
Tay-Sachs disease (TSD) is a rare neurodegenerative disorder caused by autosomal recessive mutations in the HEXA gene on chromosome 15 that encodes β-hexosaminidase. Deficiency results accumulation of GM2 ganglioside, glycosphingolipid, lysosomes. Currently, there no effective treatment for TSD. We generated induced pluripotent stem cells (iPSCs) from two TSD patient dermal fibroblast lines and further differentiated them into neural (NSCs). The exhibited phenotype lysosomal lipid...