A5060809038- CRISPR and Genetic Engineering
- Pluripotent Stem Cells Research
- Retinal Development and Disorders
- Virus-based gene therapy research
- Retinal Diseases and Treatments
- Renal and related cancers
- RNA modifications and cancer
- Platelet Disorders and Treatments
- Cancer-related gene regulation
- RNA Research and Splicing
- Retinal and Macular Surgery
- Genomics and Chromatin Dynamics
- Telomeres, Telomerase, and Senescence
- Epigenetics and DNA Methylation
- RNA Interference and Gene Delivery
- melanin and skin pigmentation
- Retinoids in leukemia and cellular processes
- Pancreatic function and diabetes
- Skin Protection and Aging
- Advanced biosensing and bioanalysis techniques
- Zebrafish Biomedical Research Applications
- Plasma Diagnostics and Applications
- Immunodeficiency and Autoimmune Disorders
- Underwater Acoustics Research
- Advanced SAR Imaging Techniques
University of California, Los Angeles
2011-2024
Century Therapeutics (United States)
2023-2024
Children's Hospital of Philadelphia
2010-2020
Penn Presbyterian Medical Center
2016-2020
University of Pennsylvania
2016-2020
Star Center
2020
California University of Pennsylvania
2015
Emory University Hospital
2013-2014
Virginia Commonwealth University Medical Center
2011
Philadelphia University
2011
The development of methods to achieve efficient reprogramming human cells while avoiding the permanent presence transgenes represents a critical step toward use induced pluripotent stem (iPSC) for clinical purposes, such as disease modeling or reconstituting therapies. Although several exist generating iPSC free from mouse neonatal normal tissues, sufficiently system is still needed widespread derivation disease-specific humans with inherited degenerative diseases. Here, we report humanized...
Patients with Down syndrome (trisomy 21, T21) have hematologic abnormalities throughout life. Newborns frequently exhibit abnormal blood counts and a clonal preleukemia. Human T21 fetal livers contain expanded erythro-megakaryocytic precursors enhanced proliferative capacity. The impact of on the earliest stages embryonic hematopoiesis is unknown nearly impossible to examine in human subjects. We modeled yolk sac using induced pluripotent stem cells (iPSCs). Blood progenitor populations...
Induced pluripotent stem cells (iPSCs) provide an inexhaustible source of for modeling disease and testing drugs. Here we develop a bioinformatic approach to detect differences between the genomic programs iPSCs derived from diseased versus normal human cohorts as they emerge during in vitro directed differentiation. Using generated cohort carrying mutations (PiZZ) gene responsible alpha-1 antitrypsin (AAT) deficiency, find that global transcriptomes PiZZ diverge controls upon...
Through the ectopic expression of four transcription factors, Oct4, Klf4, Sox2 and cMyc, human somatic cells can be converted to a pluripotent state, generating so-called induced stem (iPSCs)1-4. Patient-specific iPSCs lack ethical concerns that surround embryonic (ESCs) would bypass possible immune rejection. Thus, have attracted considerable attention for disease modeling studies, screening pharmacological compounds, regenerative therapies5. We shown generation transgene-free from patients...
Choroideremia (CHM) is an X- linked retinal degeneration that symptomatic in the 1st or 2nd decade of life causing nyctalopia and loss peripheral vision. The disease progresses through mid-life, when most patients become blind. CHM a favorable target for gene augmentation therapy, as due to function protein necessary cell health, Rab Escort Protein 1 (REP1).The cDNA can be packaged recombinant adeno-associated virus (rAAV), which has established track record human therapy studies, and,...
The Wnt gene family consists of structurally related genes encoding secreted signaling molecules that have been implicated in many developmental processes, including regulation cell fate and patterning during embryogenesis. Previously, we found is required for primitive or yolk sac-derived-erythropoiesis using the murine embryonic stem (ESC) system. Here, examine effect on formation early hematopoietic progenitors derived from human ESCs. first progenitor cells ESC system express...
Glaucoma is a group of progressive optic neuropathies that share common biological and clinical characteristics including irreversible changes to the nerve visual field loss caused by death retinal ganglion cells (RGCs). The RGCs manifests as characteristic cupping or degeneration, resulting in patients with Glaucoma. Published studies on vitro RGC differentiation from stem utilized classical signaling pathways mimicking development vivo. Although many strategies allowed for generation RGCs,...
Choroideremia (CHM) is a rare monogenic, X-linked recessive inherited retinal degeneration resulting from mutations in the Rab Escort Protein-1 (REP1) encoding CHM gene. The primary cell type leading to unknown. In this study, we explored utility of induced pluripotent stem cell-derived models pigmented epithelium (iPSC-RPE) study disease pathogenesis and potential gene-based intervention four different genetically distinct forms CHM. A number abnormal biologic, biochemical, physiologic...
Human pluripotent stem cells offer a powerful system to study human biology and disease. Here, we report both express transgenes specifically in ES cell derived hematopoietic knockdown gene expression stably throughout the differentiation of cells. We characterize CD43 promoter construct that when inserted into AAVS1 "safe harbor" locus utilizing zinc finger nuclease drives GFP from transgenic line faithfully recapitulates endogenous expression. In addition, using same targeting strategy...
Cornelia de Lange syndrome (CdLS) is a complex disorder with multiple structural and developmental defects caused by mutations in regulatory proteins involved the cohesin complex. NIPBL, protein, has been identified as critical protein responsible for orchestration of transcriptomic networks necessary embryonic development. Mutations NIPBL are majority cases CdLS. Through RNA-sequencing human induced pluripotent stem cells vitro-derived cardiomyocytes, we hundreds mRNAs, pseudogenes,...
Recombinant adeno-associated virus (rAAV), produced from a nonpathogenic parvovirus, has become an increasing popular vector for gene therapy applications in human clinical trials. However, transduction and transgene expression of rAAVs can differ across vitro ex vivo cellular strategies. This study compared 11 rAAV serotypes, carrying one reporter cassette containing cytomegalovirus immediate-early enhancer (eCMV) chicken beta actin (CBA) promoter driving the enhanced green-fluorescent...
Dyskeratosis congenita (DC) is an inherited bone marrow failure syndrome characterized by the presence of short telomeres at presentation. Mutations in ten different genes, whose products are involved telomere maintenance pathway, have been shown to cause DC. The X-linked form most common disease and caused mutations gene DKC1, encoding protein dyskerin. Dyskerin required for assembly stability telomerase also ribosomal RNA (rRNA) processing where it converts specific uridines pseudouridine....
We demonstrate that the pluripotency gene OCT4 has a role in regulating differentiation via Wnt signaling. expression levels human embryonic stem cells increases transiently during first 24 hr of vitro differentiation, with occupancy increasing at endoderm regulators such as SOX17 and FOXA2. This increased correlates loss PRC2 complex inhibitory histone mark H3K27me3. Knockdown inhibits mesendoderm formation removal H3K27me3 from promoter, suggesting acts to induce complex. Furthermore,...
Diamond Blackfan Anemia (DBA) is an inherited bone marrow failure syndrome with clinical features of red cell aplasia and variable developmental abnormalities. Most affected patients have heterozygous loss function mutations in ribosomal protein genes but the pathogenic mechanism still unknown. We generated induced pluripotent stem cells from DBA carrying RPS19 or RPL5 mutations. Transcriptome analysis revealed striking dysregulation transforming growth factor β (TGFβ) signaling pathway...
Most genetically distinct inherited retinal degenerations are primary photoreceptor degenerations. We selected a severe early onset form of Leber congenital amaurosis (LCA), caused by mutations in the gene LCA5, order to test efficacy augmentation therapy for ciliopathy. The LCA5-encoded protein, Lebercilin, is essential trafficking proteins and vesicles outer segment. Using AAV serotype AAV7m8 deliver human LCA5 cDNA into an Lca5 null mouse model we show partial rescue structure visual...
This manuscript illustrates a protocol for efficiently creating integration-free human induced pluripotent stem cells (iPSCs) from peripheral blood using episomal plasmids and histone deacetylase (HDAC) inhibitors. The advantages of this approach include: (1) the use minimal amount as source material; (2) nonintegrating reprogramming vectors; (3) cost effective method generating vector free iPSCs; (4) single transfection; (5) small molecules to facilitate epigenetic reprogramming. Briefly,...
Through the ectopic expression of four transcription factors, Oct4, Klf4, Sox2 and cMyc, human somatic cells can be converted to a pluripotent state, generating so-called induced stem (iPSCs)1-4. Patient-specific iPSCs lack ethical concerns that surround embryonic (ESCs) would bypass possible immune rejection. Thus, have attracted considerable attention for disease modeling studies, screening pharmacological compounds, regenerative therapies5. We shown generation transgene-free from patients...
Validation of gene transfer vectors containing tissue-specific promoters in cell-based functional assays poses a formidable challenge for therapy product development. Here, we describe novel approach based on CRISPR/dCas9 transcriptional activation to achieve robust transgene expression from cassettes tissue or cell type-specific after infection with AAV systems. Guide RNA sequences targeting two that are highly active within mammalian photoreceptors were screened promoter assay. Using this...