Use of genome-editing tools to treat sickle cell disease

Tasan, Ipek; Jain, Surbhi; Zhao, Huimin

doi:10.1007/s00439-016-1688-0

Cited by 27 publications

(24 citation statements)

References 169 publications

(198 reference statements)

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“…The promise of gene therapy for CAMT and other hereditary hematologic disorders awaits successes in the rapidly advancing field of genome editing of patient-derived stem cells. 17,30 Because only a subset of hematopoietic progenitor cells may need to be edited to achieve long-term polyclonal hematopoiesis, this approach could represent a cure for the majority of CAMT patients.…”

Section: Discussionmentioning

confidence: 99%

Gene editing rescue of a novel MPL mutant associated with congenital amegakaryocytic thrombocytopenia

et al. 2017

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Key Points• We report unique familial cases of CAMT presenting with a novel MPL W272R mutation in the background of the activating MPL K39N mutation.• Function of mutant Mpl receptor can be rescued using 2 approaches: autophagic cell surface delivery and CRISPR-Cas9 gene editing.Thrombopoietin (Tpo) and its receptor (Mpl) are the principal regulators of early and late thrombopoiesis and hematopoietic stem cell maintenance. Mutations in MPL can drastically impair its function and be a contributing factor in multiple hematologic malignancies, including congenital amegakaryocytic thrombocytopenia (CAMT). CAMT is characterized by severe thrombocytopenia at birth, which progresses to bone marrow failure and pancytopenia. Here we report unique familial cases of CAMT that presented with a previously unreported MPL mutation: T814C (W272R) in the background of the activating MPL G117T(K39N or Baltimore) mutation. Confocal microscopy, proliferation and surface biotinylation assays, co-immunoprecipitation, and western blotting analysis were used to elucidate the function and trafficking of Mpl mutants. Results showed that Mpl protein bearing the W272R mutation, alone or together with the K39N mutation, lacks detectable surface expression while being strongly colocalized with the endoplasmic reticulum (ER) marker calreticulin.Both WT and K39N-mutated Mpl were found to be signaling competent, but single or double mutants bearing W272R were unresponsive to Tpo. Function of the deficient Mpl receptor could be rescued by using 2 separate approaches: (1) GRASP55 overexpression, which partially restored Tpo-induced signaling of mutant Mpl by activating an autophagy-dependent secretory pathway and thus forcing ER-trapped immature receptors to traffic to the cell surface; and (2) CRISPR-Cas9 gene editing used to repair MPL T814C mutation in transfected cell lines and primary umbilical cord blood-derived CD34 1 cells. We demonstrate proof of principle for rescue of mutant Mpl function by using gene editing of primary hematopoietic stem cells, which indicates direct therapeutic applications for CAMT patients.

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Section: Discussionmentioning

confidence: 99%

Gene editing rescue of a novel MPL mutant associated with congenital amegakaryocytic thrombocytopenia

et al. 2017

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“…In fact, CRISPR/Cas9, a recently developed genome-editing tool, has been proposed as a way to cure sickle-cell anemia (Tasan et al 2016). The key is in editing the genome such that fetal hemoglobin is produced in addition to adult hemoglobin.…”

Section: Discussionmentioning

confidence: 99%

Mechanisms and causality in molecular diseases

Keenan

Shvartsman

2017

HPLS

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How is a disease contracted, and how does it progress through the body? Answers to these questions are fundamental to understanding both basic biology and medicine. Advances in the biomedical sciences continue to provide more tools to address these fundamental questions and to uncover questions that have not been thought of before. Despite these major advances, we are still facing conceptual and technical challenges when learning about the etiology of disease, especially for genetic diseases. In this review, we illustrate this point by discussing the causal links between molecular mechanisms and systems-level phenotypes in molecular diseases. We begin with an examination of sickle cell anemia, and how mechanisms of the disease have been comprehended over the last century. While sickle cell anemia involves a mutation in a single protein in a single cell type, other diseases involve mutations in networks with many protein interactions and in diverse cell types. We introduce the challenges that result from these differences and illustrate the current obstacles by discussing the RASopathies, a recently discovered class of developmental syndromes that result from mutations in signaling networks. Methods to study mutant genotypes that lead to mutant phenotypes are discussed, particularly the use of model organisms and mutant proteins to study protein interactions that may be important for development of disease. These studies will point toward the future of diagnosing and treating genetic disease.

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“…symptomatic in nature. However, gene editing empowers researchers to overcome some of the fundamental challenges inherent to the treatment of cystic fibrosis (Harrison et al 2016), hemoglobinopathies (Cottle et al 2016;Tasan et al 2016), hemophilia (Park et al 2016), and Duchenne muscular dystrophy (DMD) (Robinson-Hamm and Gersbach 2016).…”

Section: Correction Of Genetic Diseasesmentioning

confidence: 99%