Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers

Laura Breda, Irene Motta, Silvia Lourenco, Chiara Gemmo, Wulan Deng, Jeremy W. Rupon, Osheiza Y. Abdulmalik, Deepa G. Manwani, Gerd A. Blobel, Stefano Rivella

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Overcoming the silencing of the fetal γ-globin gene has been a long-standing goal in the treatment of sickle cell disease (SCD). The major transcriptional enhancer of the β-globin locus, called the locus control region (LCR), dynamically interacts with the developmental stage-appropriate β-type globin genes via chromatin looping, a process requiring the protein Ldb1. In adult erythroid cells, the LCR can be redirected from the adult β- to the fetal γ-globin promoter by tethering Ldb1 to the human γ-globin promoter with custom designed zinc finger (ZF) proteins (ZF-Ldb1), leading to reactivation of γ-globin gene expression. To compare this approach to pharmacologic reactivation of fetal hemoglobin (HbF), hematopoietic cells from patients with SCD were treated with a lentivirus expressing the ZF-Ldb1 or with chemical HbF inducers. The HbF increase in cells treated with ZF-Ldb1 was more than double that observed with decitabine and pomalidomide; butyrate had an intermediate effect whereas tranylcypromine and hydroxyurea showed relatively low HbF reactivation. ZF-Ldb1 showed comparatively little toxicity, and reduced sickle hemoglobin (HbS) synthesis as well as sickling of SCD erythroid cells under hypoxic conditions. The efficacy and low cytotoxicity of lentiviral mediated ZF-Ldb1 gene transfer compared with the drug regimens support its therapeutic potential for the treatment of SCD.

Original languageEnglish (US)
Pages (from-to)1139-1143
Number of pages5
JournalBlood
Volume128
Issue number8
DOIs
StatePublished - Aug 25 2016

Fingerprint

Fetal Hemoglobin
Globins
Zinc Fingers
Chromatin
Zinc
Sickle Cell Anemia
Locus Control Region
Erythroid Cells
decitabine
Genes
Tranylcypromine
Gene transfer
Sickle Hemoglobin
Lentivirus
Hydroxyurea
Butyrates
Cytotoxicity
Gene expression
Toxicity
Proteins

ASJC Scopus subject areas

  • Immunology
  • Biochemistry
  • Hematology
  • Cell Biology

Cite this

Breda, L., Motta, I., Lourenco, S., Gemmo, C., Deng, W., Rupon, J. W., ... Rivella, S. (2016). Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers. Blood, 128(8), 1139-1143. https://doi.org/10.1182/blood-2016-01-691089

Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers. / Breda, Laura; Motta, Irene; Lourenco, Silvia; Gemmo, Chiara; Deng, Wulan; Rupon, Jeremy W.; Abdulmalik, Osheiza Y.; Manwani, Deepa G.; Blobel, Gerd A.; Rivella, Stefano.

In: Blood, Vol. 128, No. 8, 25.08.2016, p. 1139-1143.

Research output: Contribution to journalArticle

Breda, L, Motta, I, Lourenco, S, Gemmo, C, Deng, W, Rupon, JW, Abdulmalik, OY, Manwani, DG, Blobel, GA & Rivella, S 2016, 'Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers', Blood, vol. 128, no. 8, pp. 1139-1143. https://doi.org/10.1182/blood-2016-01-691089
Breda, Laura ; Motta, Irene ; Lourenco, Silvia ; Gemmo, Chiara ; Deng, Wulan ; Rupon, Jeremy W. ; Abdulmalik, Osheiza Y. ; Manwani, Deepa G. ; Blobel, Gerd A. ; Rivella, Stefano. / Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers. In: Blood. 2016 ; Vol. 128, No. 8. pp. 1139-1143.
@article{a1a2bd331fea419985a7ca05c51a7dfb,
title = "Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers",
abstract = "Overcoming the silencing of the fetal γ-globin gene has been a long-standing goal in the treatment of sickle cell disease (SCD). The major transcriptional enhancer of the β-globin locus, called the locus control region (LCR), dynamically interacts with the developmental stage-appropriate β-type globin genes via chromatin looping, a process requiring the protein Ldb1. In adult erythroid cells, the LCR can be redirected from the adult β- to the fetal γ-globin promoter by tethering Ldb1 to the human γ-globin promoter with custom designed zinc finger (ZF) proteins (ZF-Ldb1), leading to reactivation of γ-globin gene expression. To compare this approach to pharmacologic reactivation of fetal hemoglobin (HbF), hematopoietic cells from patients with SCD were treated with a lentivirus expressing the ZF-Ldb1 or with chemical HbF inducers. The HbF increase in cells treated with ZF-Ldb1 was more than double that observed with decitabine and pomalidomide; butyrate had an intermediate effect whereas tranylcypromine and hydroxyurea showed relatively low HbF reactivation. ZF-Ldb1 showed comparatively little toxicity, and reduced sickle hemoglobin (HbS) synthesis as well as sickling of SCD erythroid cells under hypoxic conditions. The efficacy and low cytotoxicity of lentiviral mediated ZF-Ldb1 gene transfer compared with the drug regimens support its therapeutic potential for the treatment of SCD.",
author = "Laura Breda and Irene Motta and Silvia Lourenco and Chiara Gemmo and Wulan Deng and Rupon, {Jeremy W.} and Abdulmalik, {Osheiza Y.} and Manwani, {Deepa G.} and Blobel, {Gerd A.} and Stefano Rivella",
year = "2016",
month = "8",
day = "25",
doi = "10.1182/blood-2016-01-691089",
language = "English (US)",
volume = "128",
pages = "1139--1143",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "8",

}

TY - JOUR

T1 - Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers

AU - Breda, Laura

AU - Motta, Irene

AU - Lourenco, Silvia

AU - Gemmo, Chiara

AU - Deng, Wulan

AU - Rupon, Jeremy W.

AU - Abdulmalik, Osheiza Y.

AU - Manwani, Deepa G.

AU - Blobel, Gerd A.

AU - Rivella, Stefano

PY - 2016/8/25

Y1 - 2016/8/25

N2 - Overcoming the silencing of the fetal γ-globin gene has been a long-standing goal in the treatment of sickle cell disease (SCD). The major transcriptional enhancer of the β-globin locus, called the locus control region (LCR), dynamically interacts with the developmental stage-appropriate β-type globin genes via chromatin looping, a process requiring the protein Ldb1. In adult erythroid cells, the LCR can be redirected from the adult β- to the fetal γ-globin promoter by tethering Ldb1 to the human γ-globin promoter with custom designed zinc finger (ZF) proteins (ZF-Ldb1), leading to reactivation of γ-globin gene expression. To compare this approach to pharmacologic reactivation of fetal hemoglobin (HbF), hematopoietic cells from patients with SCD were treated with a lentivirus expressing the ZF-Ldb1 or with chemical HbF inducers. The HbF increase in cells treated with ZF-Ldb1 was more than double that observed with decitabine and pomalidomide; butyrate had an intermediate effect whereas tranylcypromine and hydroxyurea showed relatively low HbF reactivation. ZF-Ldb1 showed comparatively little toxicity, and reduced sickle hemoglobin (HbS) synthesis as well as sickling of SCD erythroid cells under hypoxic conditions. The efficacy and low cytotoxicity of lentiviral mediated ZF-Ldb1 gene transfer compared with the drug regimens support its therapeutic potential for the treatment of SCD.

AB - Overcoming the silencing of the fetal γ-globin gene has been a long-standing goal in the treatment of sickle cell disease (SCD). The major transcriptional enhancer of the β-globin locus, called the locus control region (LCR), dynamically interacts with the developmental stage-appropriate β-type globin genes via chromatin looping, a process requiring the protein Ldb1. In adult erythroid cells, the LCR can be redirected from the adult β- to the fetal γ-globin promoter by tethering Ldb1 to the human γ-globin promoter with custom designed zinc finger (ZF) proteins (ZF-Ldb1), leading to reactivation of γ-globin gene expression. To compare this approach to pharmacologic reactivation of fetal hemoglobin (HbF), hematopoietic cells from patients with SCD were treated with a lentivirus expressing the ZF-Ldb1 or with chemical HbF inducers. The HbF increase in cells treated with ZF-Ldb1 was more than double that observed with decitabine and pomalidomide; butyrate had an intermediate effect whereas tranylcypromine and hydroxyurea showed relatively low HbF reactivation. ZF-Ldb1 showed comparatively little toxicity, and reduced sickle hemoglobin (HbS) synthesis as well as sickling of SCD erythroid cells under hypoxic conditions. The efficacy and low cytotoxicity of lentiviral mediated ZF-Ldb1 gene transfer compared with the drug regimens support its therapeutic potential for the treatment of SCD.

UR - http://www.scopus.com/inward/record.url?scp=84996807220&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84996807220&partnerID=8YFLogxK

U2 - 10.1182/blood-2016-01-691089

DO - 10.1182/blood-2016-01-691089

M3 - Article

VL - 128

SP - 1139

EP - 1143

JO - Blood

JF - Blood

SN - 0006-4971

IS - 8

ER -