Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.

<h4>Background</h4>Sickle cell disease (SCD), a congenital hemolytic anemia that exacts terrible global morbidity and mortality, is driven by polymerization of mutated sickle hemoglobin (HbS) in red blood cells (RBCs). Fetal hemoglobin (HbF) interferes with this polymerization, but HbF i...

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Main Authors: Robert Molokie, Donald Lavelle, Michel Gowhari, Michael Pacini, Lani Krauz, Johara Hassan, Vinzon Ibanez, Maria A Ruiz, Kwok Peng Ng, Philip Woost, Tomas Radivoyevitch, Daisy Pacelli, Sherry Fada, Matthew Rump, Matthew Hsieh, John F Tisdale, James Jacobberger, Mitch Phelps, James Douglas Engel, Santhosh Saraf, Lewis L Hsu, Victor Gordeuk, Joseph DeSimone, Yogen Saunthararajah
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-09-01
Series:PLoS Medicine
Online Access:https://journals.plos.org/plosmedicine/article/file?id=10.1371/journal.pmed.1002382&type=printable
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author Robert Molokie
Donald Lavelle
Michel Gowhari
Michael Pacini
Lani Krauz
Johara Hassan
Vinzon Ibanez
Maria A Ruiz
Kwok Peng Ng
Philip Woost
Tomas Radivoyevitch
Daisy Pacelli
Sherry Fada
Matthew Rump
Matthew Hsieh
John F Tisdale
James Jacobberger
Mitch Phelps
James Douglas Engel
Santhosh Saraf
Lewis L Hsu
Victor Gordeuk
Joseph DeSimone
Yogen Saunthararajah
author_facet Robert Molokie
Donald Lavelle
Michel Gowhari
Michael Pacini
Lani Krauz
Johara Hassan
Vinzon Ibanez
Maria A Ruiz
Kwok Peng Ng
Philip Woost
Tomas Radivoyevitch
Daisy Pacelli
Sherry Fada
Matthew Rump
Matthew Hsieh
John F Tisdale
James Jacobberger
Mitch Phelps
James Douglas Engel
Santhosh Saraf
Lewis L Hsu
Victor Gordeuk
Joseph DeSimone
Yogen Saunthararajah
author_sort Robert Molokie
collection DOAJ
description <h4>Background</h4>Sickle cell disease (SCD), a congenital hemolytic anemia that exacts terrible global morbidity and mortality, is driven by polymerization of mutated sickle hemoglobin (HbS) in red blood cells (RBCs). Fetal hemoglobin (HbF) interferes with this polymerization, but HbF is epigenetically silenced from infancy onward by DNA methyltransferase 1 (DNMT1).<h4>Methods and findings</h4>To pharmacologically re-induce HbF by DNMT1 inhibition, this first-in-human clinical trial (NCT01685515) combined 2 small molecules-decitabine to deplete DNMT1 and tetrahydrouridine (THU) to inhibit cytidine deaminase (CDA), the enzyme that otherwise rapidly deaminates/inactivates decitabine, severely limiting its half-life, tissue distribution, and oral bioavailability. Oral decitabine doses, administered after oral THU 10 mg/kg, were escalated from a very low starting level (0.01, 0.02, 0.04, 0.08, or 0.16 mg/kg) to identify minimal doses active in depleting DNMT1 without cytotoxicity. Patients were SCD adults at risk of early death despite standard-of-care, randomized 3:2 to THU-decitabine versus placebo in 5 cohorts of 5 patients treated 2X/week for 8 weeks, with 4 weeks of follow-up. The primary endpoint was ≥ grade 3 non-hematologic toxicity. This endpoint was not triggered, and adverse events (AEs) were not significantly different in THU-decitabine-versus placebo-treated patients. At the decitabine 0.16 mg/kg dose, plasma concentrations peaked at approximately 50 nM (Cmax) and remained elevated for several hours. This dose decreased DNMT1 protein in peripheral blood mononuclear cells by >75% and repetitive element CpG methylation by approximately 10%, and increased HbF by 4%-9% (P < 0.001), doubling fetal hemoglobin-enriched red blood cells (F-cells) up to approximately 80% of total RBCs. Total hemoglobin increased by 1.2-1.9 g/dL (P = 0.01) as reticulocytes simultaneously decreased; that is, better quality and efficiency of HbF-enriched erythropoiesis elevated hemoglobin using fewer reticulocytes. Also indicating better RBC quality, biomarkers of hemolysis, thrombophilia, and inflammation (LDH, bilirubin, D-dimer, C-reactive protein [CRP]) improved. As expected with non-cytotoxic DNMT1-depletion, platelets increased and neutrophils concurrently decreased, but not to an extent requiring treatment holds. As an early phase study, limitations include small patient numbers at each dose level and narrow capacity to evaluate clinical benefits.<h4>Conclusion</h4>Administration of oral THU-decitabine to patients with SCD was safe in this study and, by targeting DNMT1, upregulated HbF in RBCs. Further studies should investigate clinical benefits and potential harms not identified to date.<h4>Trial registration</h4>ClinicalTrials.gov, NCT01685515.
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spelling doaj-art-c7733c856c0b41248e7cccfad4c6af2d2025-08-20T03:11:25ZengPublic Library of Science (PLoS)PLoS Medicine1549-12771549-16762017-09-01149e100238210.1371/journal.pmed.1002382Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.Robert MolokieDonald LavelleMichel GowhariMichael PaciniLani KrauzJohara HassanVinzon IbanezMaria A RuizKwok Peng NgPhilip WoostTomas RadivoyevitchDaisy PacelliSherry FadaMatthew RumpMatthew HsiehJohn F TisdaleJames JacobbergerMitch PhelpsJames Douglas EngelSanthosh SarafLewis L HsuVictor GordeukJoseph DeSimoneYogen Saunthararajah<h4>Background</h4>Sickle cell disease (SCD), a congenital hemolytic anemia that exacts terrible global morbidity and mortality, is driven by polymerization of mutated sickle hemoglobin (HbS) in red blood cells (RBCs). Fetal hemoglobin (HbF) interferes with this polymerization, but HbF is epigenetically silenced from infancy onward by DNA methyltransferase 1 (DNMT1).<h4>Methods and findings</h4>To pharmacologically re-induce HbF by DNMT1 inhibition, this first-in-human clinical trial (NCT01685515) combined 2 small molecules-decitabine to deplete DNMT1 and tetrahydrouridine (THU) to inhibit cytidine deaminase (CDA), the enzyme that otherwise rapidly deaminates/inactivates decitabine, severely limiting its half-life, tissue distribution, and oral bioavailability. Oral decitabine doses, administered after oral THU 10 mg/kg, were escalated from a very low starting level (0.01, 0.02, 0.04, 0.08, or 0.16 mg/kg) to identify minimal doses active in depleting DNMT1 without cytotoxicity. Patients were SCD adults at risk of early death despite standard-of-care, randomized 3:2 to THU-decitabine versus placebo in 5 cohorts of 5 patients treated 2X/week for 8 weeks, with 4 weeks of follow-up. The primary endpoint was ≥ grade 3 non-hematologic toxicity. This endpoint was not triggered, and adverse events (AEs) were not significantly different in THU-decitabine-versus placebo-treated patients. At the decitabine 0.16 mg/kg dose, plasma concentrations peaked at approximately 50 nM (Cmax) and remained elevated for several hours. This dose decreased DNMT1 protein in peripheral blood mononuclear cells by >75% and repetitive element CpG methylation by approximately 10%, and increased HbF by 4%-9% (P < 0.001), doubling fetal hemoglobin-enriched red blood cells (F-cells) up to approximately 80% of total RBCs. Total hemoglobin increased by 1.2-1.9 g/dL (P = 0.01) as reticulocytes simultaneously decreased; that is, better quality and efficiency of HbF-enriched erythropoiesis elevated hemoglobin using fewer reticulocytes. Also indicating better RBC quality, biomarkers of hemolysis, thrombophilia, and inflammation (LDH, bilirubin, D-dimer, C-reactive protein [CRP]) improved. As expected with non-cytotoxic DNMT1-depletion, platelets increased and neutrophils concurrently decreased, but not to an extent requiring treatment holds. As an early phase study, limitations include small patient numbers at each dose level and narrow capacity to evaluate clinical benefits.<h4>Conclusion</h4>Administration of oral THU-decitabine to patients with SCD was safe in this study and, by targeting DNMT1, upregulated HbF in RBCs. Further studies should investigate clinical benefits and potential harms not identified to date.<h4>Trial registration</h4>ClinicalTrials.gov, NCT01685515.https://journals.plos.org/plosmedicine/article/file?id=10.1371/journal.pmed.1002382&type=printable
spellingShingle Robert Molokie
Donald Lavelle
Michel Gowhari
Michael Pacini
Lani Krauz
Johara Hassan
Vinzon Ibanez
Maria A Ruiz
Kwok Peng Ng
Philip Woost
Tomas Radivoyevitch
Daisy Pacelli
Sherry Fada
Matthew Rump
Matthew Hsieh
John F Tisdale
James Jacobberger
Mitch Phelps
James Douglas Engel
Santhosh Saraf
Lewis L Hsu
Victor Gordeuk
Joseph DeSimone
Yogen Saunthararajah
Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.
PLoS Medicine
title Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.
title_full Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.
title_fullStr Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.
title_full_unstemmed Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.
title_short Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study.
title_sort oral tetrahydrouridine and decitabine for non cytotoxic epigenetic gene regulation in sickle cell disease a randomized phase 1 study
url https://journals.plos.org/plosmedicine/article/file?id=10.1371/journal.pmed.1002382&type=printable
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