Lentiviral Transfer of γ-Globin with Fusion Gene NUP98-HOXA10HD Expands Hematopoietic Stem Cells and Ameliorates Murine β-Thalassemia

Zhao, Hui; Abraham, Allistair; Kim, Yoon-Sang; Wang, Yongdong; Pestina, Tamara I.; Zhan, Jun; Humphries, R. Keith; Nienhuis, Arthur W.; Persons, Derek A.

doi:10.1016/j.ymthe.2017.01.019

Cited by 6 publications

(5 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results align with previous reports presenting HbF induction in conditions leading to rapid erythroid regeneration, such as transient erythroblastogenia of childhood [13], erythropoietin treatment of baboons [14], and bone marrow transplantation [15]. Because elevated HbF levels reduce the complications and severity of SCD and β-thalassemia, γ-globin gene addition [16] and targeted induction strategies [5] have been evaluated in preclinical models and are currently being investigated in clinical trials (ClinicalTrials.gov Identifiers: NCT02186418 and NCT03282656). On the basis of the data presented here and in previously published works, the results of these approaches should be evaluated after hematopoietic development has stabilized to have long-term clinical benefit.…”

Section: Discussionsupporting

confidence: 87%

Fetal hemoglobin and F-cell variance in mobilized CD34+ cell-transplanted rhesus monkeys

Demirci¹,

Haro‐Mora²,

Yapundich³

et al. 2019

Experimental Hematology

View full text Add to dashboard Cite

Elevated fetal hemoglobin (HbF) is associated with reduced severity of sickle cell disease. Therefore, γ-globin protein levels and F-cell (HbF-positive red blood cell) percentages are used for estimation of clinical benefit. Here, we monitored transplantation-related changes in HbF and F-cell percentages for rhesus macaques (Macaca mulatta) following total body irradiation or busulfan conditioning prior to CD34 + cell transplantation. HbF protein expression peaked in the first 4-9 weeks posttransplant (0.99%-2.53%), and F-cells increased in the first 6-17 weeks posttransplant (8.7%-45.3%). HbF and F-cell ratios gradually decreased and stabilized to levels similar to those of control animals (1.96 ± 1.97% for F cells and 0.49 ± 0.19% γ-globin expression) 4-7 months post-transplant. These findings confirm and expand on previous reports of transient induction in HbF and F-cell percentages in rhesus macaques following CD34 + cell transplantation, an observation that must be taken into consideration when evaluating therapeutic strategies that aim to specifically elevate HbF expression, which are currently in clinical development. Published by Elsevier Inc. on behalf of ISEH -Society for Hematology and Stem Cells.Sickle cell disease (SCD) is a monogenic disorder caused by a missense mutation in the β-globin gene, which results in the production of sickle hemoglobin (HbS, α 2 βS 2 ) that polymerizes under hypoxic conditions. Sickle erythrocytes occlude small blood vessels leading to ischemic tissue damage, resulting in severe pain and/or organ failure [1]. High fetal hemoglobin (HbF, α 2 γ 2 ) levels are clinically beneficial for SCD patients as HbF inhibits deoxy HbS polymerization. Patients with hereditary persistence of fetal hemoglobin (HPFH) mutations and a typical evenly distributed 30% HbF expression are clinically asymptomatic despite having high HbS levels [2]. Additionally, prolonged survival [3] and lower pain frequency [4] have been reported in patients with elevated HbF levels.

show abstract

Section: Discussionsupporting

confidence: 87%

Fetal hemoglobin and F-cell variance in mobilized CD34+ cell-transplanted rhesus monkeys

Demirci¹,

Haro‐Mora²,

Yapundich³

et al. 2019

Experimental Hematology

View full text Add to dashboard Cite

show abstract

“…www.nature.com/scientificreports www.nature.com/scientificreports/ co-inheritance of HPFH with β-thalassemia was noticed to alleviate the clinical manifestations of the latter 14 . Therefore, to attain a clinical profit for β-hemoglobinopathies, several attempts were made to re-establish the expression of HbF either by lentiviral transfer of the γ-globin gene or by CRISPR/Cas9-mediated gene disruption of specific regulators [14][15][16][17][18][19] .…”

mentioning

confidence: 99%

Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34+ HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin

Lamsfus‐Calle

Daniel‐Moreno

Antony

et al. 2020

Sci Rep

View full text Add to dashboard Cite

β-hemoglobinopathies are caused by abnormal or absent production of hemoglobin in the blood due to mutations in the β-globin gene (HBB). Imbalanced expression of adult hemoglobin (HbA) induces strong anemia in patients suffering from the disease. However, individuals with natural-occurring mutations in the HBB cluster or related genes, compensate this disparity through γ-globin expression and subsequent fetal hemoglobin (HbF) production. Several preclinical and clinical studies have been performed in order to induce HbF by knocking-down genes involved in HbF repression (KLF1 and BCL11A) or disrupting the binding sites of several transcription factors in the γ-globin gene (HBG1/2). In this study, we thoroughly compared the different CRISPR/Cas9 gene-disruption strategies by gene editing analysis and assessed their safety profile by RNA-seq and GUIDE-seq. All approaches reached therapeutic levels of HbF after gene editing and showed similar gene expression to the control sample, while no significant off-targets were detected by GUIDE-seq. Likewise, all three gene editing platforms were established in the GMP-grade CliniMACS Prodigy, achieving similar outcome to preclinical devices. Based on this gene editing comparative analysis, we concluded that BCL11A is the most clinically relevant approach while HBG1/2 could represent a promising alternative for the treatment of β-hemoglobinopathies. Sickle cell disease (SCD) and β-thalassemia, commonly known as β-hemoglobinopathies, are inherited blood disorders caused by mutations in the human β-globin gene (HBB) 1-4. In healthy condition, adult human hemoglobin (HbA) consists of 2 α and 2 β chains, whereas fetal hemoglobin (HbF) expressed in early gestation comprises 2 α chains and 2 γ chains. Notably, HbF was observed to bind oxygen with greater affinity than HbA, being functional when reactivated in adults 3,5,6. Recent studies have generated substantial experimental evidence that HbF reactivation by gene disruption of specific transcription factors and regulators could provide a therapeutic benefit for β-hemoglobinopathies 7. It has long been appreciated that KLF1 and BCL11A are key regulators involved in the process of γto β-globin switching and the repression of these genes leads to HbF resurgence 6-11. Interestingly, healthy individuals with a benign genetic condition namely hereditary persistence of fetal hemoglobin (HPFH) were observed to exhibit persistent production of functional HbF 4,10,12,13. HPFH is caused by large deletions in the δand β-globin genes, or point mutations in the γ-globin promoter and γ-globin repressors, such as KLF1 and BCL11A 5. Importantly,

show abstract

“…The Nup98 fusion protein was able to increase the number of human hematopoietic stem cells 4 months after initial transfer into immunodeficient mouse hosts [171]. This strategy has been shown to be effective in the treatment of a murine model of Beta Thalassemia [172]. …”

Section: Npcs In Other Disordersmentioning

confidence: 99%

The roles of the nuclear pore complex in cellular dysfunction, aging and disease

Sakuma

D’Angelo

2017

Seminars in Cell & Developmental Biology

110

View full text Add to dashboard Cite

The study of the Nuclear Pore Complex (NPC), the proteins that compose it (nucleoporins), and the nucleocytoplasmic transport that it controls has revealed an unexpected layer to pathogenic disease onset and progression. Recent advances in the study of the regulation of NPC composition and function suggest that the precise control of this structure is necessary to prevent diseases from arising or progressing. Here we discuss the role of nucleoporins in a diverse set of diseases, many of which directly or indirectly increase in occurrence and severity as we age, and often shorten the human lifespan. NPC biology has been shown to play a direct role in these diseases and therefore in the process of healthy aging.

show abstract

Lentiviral Transfer of γ-Globin with Fusion Gene NUP98-HOXA10HD Expands Hematopoietic Stem Cells and Ameliorates Murine β-Thalassemia

Cited by 6 publications

References 36 publications

Fetal hemoglobin and F-cell variance in mobilized CD34+ cell-transplanted rhesus monkeys

Fetal hemoglobin and F-cell variance in mobilized CD34+ cell-transplanted rhesus monkeys

Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34+ HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin

The roles of the nuclear pore complex in cellular dysfunction, aging and disease

Contact Info

Product

Resources

About