338. Evaluation of TALENs and the CRISPR/Cas9 Nuclease System To Correct the Sickle Cell Disease Mutation

Hoban, Megan D.; Lumaquin, Dianne; Kuo, Caroline Y.; Romero, Zulema; Young, Courtney S.; Ho, Michelle; Long, Joseph; Coss, Nathan; Bjurström, Carmen Flores; Mojadidi, Michelle; Hollis, Roger P.; Kohn, Donald B.

doi:10.1016/s1525-0016(16)33947-8

Cited by 7 publications

(8 citation statements)

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“…However, concerns regarding the safety of genome editing, particularly upon delivery in vivo or transplantation must be taken seriously. Thus, this is currently under major consideration, and several studies have recently demonstrated the effectiveness and, particularly, safety of genome editing in a broad spectrum of hematological and coagulation diseases (Hoban et al, 2016;Ou et al, 2016;Ohmori et al, 2017;Lyu et al, 2018;Ramaswamy et al, 2018;Cosenza et al, 2021;Son et al, 2022). Importantly, clinical trials implementing CRISPR-edited HSPCs for the treatment of severe Sickle Cell Disease (SCD) and β-thalassemia are currently on-going, with no reported off-target effects (Frangoul et al, 2021).…”

Section: The Future Of Liver Organoid-based Therapiesmentioning

confidence: 99%

“iPSC-derived liver organoids and inherited bleeding disorders: Potential and future perspectives”

Roman

Stavik²,

Lauritzen³

et al. 2023

Front. Physiol.

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The bleeding phenotype of hereditary coagulation disorders is caused by the low or undetectable activity of the proteins involved in hemostasis, due to a broad spectrum of genetic alterations. Most of the affected coagulation factors are produced in the liver. Therefore, two-dimensional (2D) cultures of primary human hepatocytes and recombinant overexpression of the factors in non-human cell lines have been primarily used to mimic disease pathogenesis and as a model for innovative therapeutic strategies. However, neither human nor animal cells fully represent the hepatocellular biology and do not harbor the exact genetic background of the patient. As a result, the inability of the current in vitro models in recapitulating the in vivo situation has limited the studies of these inherited coagulation disorders. Induced Pluripotent Stem Cell (iPSC) technology offers a possible solution to overcome these limitations by reprogramming patient somatic cells into an embryonic-like pluripotent state, thus giving the possibility of generating an unlimited number of liver cells needed for modeling or therapeutic purposes. By combining this potential and the recent advances in the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology, it allows for the generation of autologous and gene corrected liver cells in the form of three-dimensional (3D) liver organoids. The organoids recapitulate cellular composition and organization of the liver, providing a more physiological model to study the biology of coagulation proteins and modeling hereditary coagulation disorders. This advanced methodology can pave the way for the development of cell-based therapeutic approaches to treat inherited coagulation disorders. In this review we will explore the use of liver organoids as a state-of-the-art methodology for modeling coagulation factors disorders and the possibilities of using organoid technology to treat the disease.

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Section: The Future Of Liver Organoid-based Therapiesmentioning

confidence: 99%

“iPSC-derived liver organoids and inherited bleeding disorders: Potential and future perspectives”

Roman

Stavik²,

Lauritzen³

et al. 2023

Front. Physiol.

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“…Genome editing, since it was pioneered in the previous century, is developing meteorically as a revolutionary therapeutic tool for genetic defects, including hematological disorders (Xie et al, 2014;Hoban et al, 2016;De Ravin et al, 2017;Orkin and Bauer, 2019) CRISPR/Cas9, a part of the bacterial acquired immune system, was adapted as a breakthrough genome engineering technology and has since been extensively used to engineer eukaryotic cells in basic research and holds great potential for gene therapy (Gasiunas et al, 2012;Jinek et al, 2012;Cong et al, 2013;Barrangou and Doudna, 2016). CRISPR/Cas9 mediated genome editing relies on sequence specific guide RNAs that assemble with Cas9 protein to create double strand breaks (DSBs) in the targeted sequence.…”

Section: Genome Editing: a Cure For Gata2 Haploinsufficiencies? Gata2mentioning

confidence: 99%

From Basic Biology to Patient Mutational Spectra of GATA2 Haploinsufficiencies: What Are the Mechanisms, Hurdles, and Prospects of Genome Editing for Treatment

Koyunlar

Pater

2020

Front. Genome Ed.

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Inherited bone marrow failure syndromes (IBMFS) are monogenetic disorders that result in a reduction of mature blood cell formation and predisposition to leukemia. In children with myeloid leukemia the gene most often mutated is Gata binding protein 2 (GATA2) and 80% of patients with GATA2 mutations develop myeloid malignancy before the age of forty. Although GATA2 is established as one of the key regulators of embryonic and adult hematopoiesis, the mechanisms behind the leukemia predisposition in GATA2 haploinsufficiencies is ambiguous. The only curative treatment option currently available is allogeneic hematopoietic stem cell transplantation (allo-SCT). However, allo-SCT can only be applied at a relatively late stage of the disease as its applicability is compromised by treatment related morbidity and mortality (TRM). Alternatively, autologous hematopoietic stem cell transplantation (auto-SCT), which is associated with significantly less TRM, might become a treatment option if repaired hematopoietic stem cells would be available. Here we discuss the recent literature on leukemia predisposition syndromes caused by GATA2 mutations, current knowledge on the function of GATA2 in the hematopoietic system and advantages and pitfalls of potential treatment options provided by genome editing.

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“…K562 cells were modified to contain sickle cell disease-causing mutation, as described previously (Hoban et al, 2016). K562 BFP cells were modified to contain monoallelic copy of the BFP gene, as described in Richardson et al (2016).…”

Section: K562 Cellsmentioning

confidence: 99%

“…Analysis of sequencing data was performed as described elsewhere (Hoban et al, 2015(Hoban et al, , 2016Lomova et al, 2019). Percentage of HDR was calculated as the (number of sequence reads containing a sickle change)/(total reads for that sample) * 100.…”

Section: Sequencing Analysis and Calculationsmentioning

confidence: 99%

Global and Local Manipulation of DNA Repair Mechanisms to Alter Site-Specific Gene Editing Outcomes in Hematopoietic Stem Cells

et al. 2020

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Monogenic disorders of the blood system have the potential to be treated by autologous stem cell transplantation of ex vivo genetically modified hematopoietic stem and progenitor cells (HSPCs). The sgRNA/Cas9 system allows for precise modification of the genome at single nucleotide resolution. However, the system is reliant on endogenous cellular DNA repair mechanisms to mend a Cas9-induced double stranded break (DSB), either by the non-homologous end joining (NHEJ) pathway or by the cell-cycle regulated homology-directed repair (HDR) pathway. Here, we describe a panel of ectopically expressed DNA repair factors and Cas9 variants assessed for their ability to promote gene correction by HDR or inhibit gene disruption by NHEJ at the HBB locus. Although transient global overexpression of DNA repair factors did not improve the frequency of gene correction in primary HSPCs, localization of factors to the DSB by fusion to the Cas9 protein did alter repair outcomes toward microhomology-mediated end joining (MMEJ) repair, an HDR event. This strategy may be useful when predictable gene editing outcomes are imperative for therapeutic success.

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338. Evaluation of TALENs and the CRISPR/Cas9 Nuclease System To Correct the Sickle Cell Disease Mutation

Cited by 7 publications

References 0 publications

“iPSC-derived liver organoids and inherited bleeding disorders: Potential and future perspectives”

“iPSC-derived liver organoids and inherited bleeding disorders: Potential and future perspectives”

From Basic Biology to Patient Mutational Spectra of GATA2 Haploinsufficiencies: What Are the Mechanisms, Hurdles, and Prospects of Genome Editing for Treatment

Global and Local Manipulation of DNA Repair Mechanisms to Alter Site-Specific Gene Editing Outcomes in Hematopoietic Stem Cells

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