Genomic discovery of potent chromatin insulators for human gene therapy

Liu, Mingdong; Maurano, Matthew T.; Wang, Hao; Qi, Heyuan; Song, Chao; Navas, Patrick A.; Emery, David W.; Stamatoyannopoulos, J; Stamatoyannopoulos, George

doi:10.1038/nbt.3062

Cited by 110 publications

(126 citation statements)

References 50 publications

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“…Evaluation of CTCF-binding sites in the human genome has established hundreds of CTCF-binding domains, so designing repeated CTCF elements with potent insulating activity that do not reduce vector titers may be possible. [62][63][64][65] Only the FV vectors with 650cHS4 in the reverse orientation fulfilled the criteria of significant insulating activity, high titer, and high fidelity of insulator transfer from vector plasmid to vector provirus. By using vectors with a strong SFFV internal promoter, we showed in the established and sensitive 32D assay that the FV vectors were significantly less genotoxic than a GV vector (CL-SGN) and similar in genotoxicity to an LV vector (LV-SFFVEGFP) (Fig.…”

Section: Discussionmentioning

confidence: 99%

Insulated Foamy Viral Vectors

et al. 2016

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Retroviral vector-mediated gene therapy is promising, but genotoxicity has limited its use in the clinic. Genotoxicity is highly dependent on the retroviral vector used, and foamy viral (FV) vectors appear relatively safe. However, internal promoters may still potentially activate nearby genes. We developed insulated FV vectors, using four previously described insulators: a version of the well-studied chicken hypersensitivity site 4 insulator (650cHS4), two synthetic CCCTC-binding factor (CTCF)-based insulators, and an insulator based on the CCAAT box-binding transcription factor/nuclear factor I (7xCTF/NF1). We directly compared these insulators for enhancer-blocking activity, effect on FV vector titer, and fidelity of transfer to both proviral long terminal repeats. The synthetic CTCF-based insulators had the strongest insulating activity, but reduced titers significantly. The 7xCTF/NF1 insulator did not reduce titers but had weak insulating activity. The 650cHS4-insulated FV vector was identified as the overall most promising vector. Uninsulated and 650cHS4-insulated FV vectors were both significantly less genotoxic than gammaretroviral vectors. Integration sites were evaluated in cord blood CD34+ cells and the 650cHS4-insulated FV vector had fewer hotspots compared with an uninsulated FV vector. These data suggest that insulated FV vectors are promising for hematopoietic stem cell gene therapy.

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Section: Discussionmentioning

confidence: 99%

Insulated Foamy Viral Vectors

et al. 2016

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“…Studies aimed at characterizing and including insulator elements into viral vectors to reduce genome toxicity are being actively pursued. 168 In addition, new technologies to genetically modify HSCs and induce pluripotent stem cells by genome editing are also being explored (see "Genome editing").…”

Section: Gene Therapymentioning

confidence: 99%

-thalassemias: paradigmatic diseases for scientific discoveries and development of innovative therapies

2015

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b-thalassemias are monogenic disorders characterized by defective synthesis of the b-globin chain, one of the major components of adult hemoglobin. A large number of mutations in the b-globin gene or its regulatory elements have been associated with b-thalassemias. Due to the complexity of the regulation of the b-globin gene and the role of red cells in many physiological processes, patients can manifest a large spectrum of phenotypes, and clinical requirements vary from patient to patient. It is important to consider the major differences in the light of potential novel therapeutics. This review summarizes the main discoveries and mechanisms associated with the synthesis of b-globin and abnormal erythropoiesis, as well as current and novel therapies. ABSTRACTThe complex phenotype of b-thalassemias b-thalassemias are monogenic disorders characterized by reduced or no synthesis of the b-globin chain, one of the major components of adult hemoglobin (HbA). Several hundred mutations in the b-globin gene or regulatory elements have been associated with b-thalassemias.

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“…The basis of TAD loops is highly enriched for CCCTCF-binding factor (CTCF) [47] (Figure 2). CTCF is known as a transcriptional regulator that functionally segregates chromosomal TADs by inhibiting enhancer-promoter interactions [49]. Importantly, the majority of mammalian TAD loops are flanked by a pair of convergent CTCF motifs that mark the TAD's left and right boundaries [50].…”

Section: Chromosome Architecture and Epigenetic Control Of Glucocortimentioning

confidence: 99%

Twenty-First Century Glucocorticoid Receptor Molecular Biology

Wang¹,

Oldenkamp²,

Oellers³

et al. 2018

Corticosteroids

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Glucocorticoids are central to homeostasis as a function of the circadian cycle, temporally preceding circulating adrenaline concentration circadian fluctuations. Virtually, all cell types express the glucocorticoid receptor (GR). GR is a transcription factor that activates gene expression by binding to enhancers. Intriguingly, not all cell types respond to GR stimulation in the same fashion at the molecular level. This indicates that GR activity is subject to epigenetic control. We discuss the molecular basis for epigenetic control of GR action at the genomic level, including the concept of topologically associating domains which may restrain the roaming range of distal enhancers. Furthermore, much evidence indicates that GR can repress gene expression programs. We therefore discuss current concepts of the molecular basis of GR-mediated gene expression repression, including non-genomic mechanisms that involve mRNA destabilization.

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Genomic discovery of potent chromatin insulators for human gene therapy

Cited by 110 publications

References 50 publications

Insulated Foamy Viral Vectors

Insulated Foamy Viral Vectors

-thalassemias: paradigmatic diseases for scientific discoveries and development of innovative therapies

Twenty-First Century Glucocorticoid Receptor Molecular Biology

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