A novel imaging flow cytometry method for the detection of histone H4 acetylation in myeloid cells.
Acetylation
Analysis of Variance
Antineoplastic Agents
/ administration & dosage
Child
Dose-Response Relationship, Drug
Ependymoma
/ drug therapy
Flow Cytometry
/ methods
Healthy Volunteers
Histone Deacetylase Inhibitors
/ administration & dosage
Histones
/ metabolism
Humans
Leukocytes
/ chemistry
Myeloid Cells
/ chemistry
Valproic Acid
/ administration & dosage
Young Adult
biomarkers
cancer
histone H4 acetylation
imaging flow cytometry
valproic acid
Journal
European journal of clinical investigation
ISSN: 1365-2362
Titre abrégé: Eur J Clin Invest
Pays: England
ID NLM: 0245331
Informations de publication
Date de publication:
Jul 2019
Jul 2019
Historique:
received:
27
11
2018
revised:
05
03
2019
accepted:
26
03
2019
pubmed:
1
4
2019
medline:
7
1
2020
entrez:
1
4
2019
Statut:
ppublish
Résumé
The histone deacetylase inhibitor (HDACI) valproic acid has been shown to inhibit the growth of multiple paediatric tumour types and is well tolerated in a childhood cancer setting. The current study was designed to develop a novel imaging flow cytometry method for the detection of histone H4 acetylation in white blood cells obtained from childhood cancer patients treated with valproic acid. HL-60 cells and whole blood samples from healthy volunteers were incubated with valproic acid (0-8 mM) for 0-24 hours, with additional blood samples collected from ependymoma patients receiving valproic acid on the SIOP Ependymoma II clinical trial. An imaging flow cytometry method was developed using an ImageStream The mean percentage of acH4-positive cells increased to a greater extent than increases in mean and median fluorescence intensity following incubation with valproic acid. Comparable results were observed for in vitro and ex vivo experiments, and the assay was shown to be appropriate for clinical sample analysis. Myeloid cells exhibited a smaller proportion of acH4-positive cells than the lymphoid population, but a greater fold increase above basal levels. The percentage of acH4-positive myeloid cells has the potential to be used as a robust pharmacodynamic biomarker for the measurement of acH4 for HDACIs. The developed assay is now being utilised in a clinical trial involving the treatment of childhood ependymoma patients with valproic acid.
Sections du résumé
BACKGROUND
BACKGROUND
The histone deacetylase inhibitor (HDACI) valproic acid has been shown to inhibit the growth of multiple paediatric tumour types and is well tolerated in a childhood cancer setting. The current study was designed to develop a novel imaging flow cytometry method for the detection of histone H4 acetylation in white blood cells obtained from childhood cancer patients treated with valproic acid.
MATERIALS AND METHODS
METHODS
HL-60 cells and whole blood samples from healthy volunteers were incubated with valproic acid (0-8 mM) for 0-24 hours, with additional blood samples collected from ependymoma patients receiving valproic acid on the SIOP Ependymoma II clinical trial. An imaging flow cytometry method was developed using an ImageStream
RESULTS
RESULTS
The mean percentage of acH4-positive cells increased to a greater extent than increases in mean and median fluorescence intensity following incubation with valproic acid. Comparable results were observed for in vitro and ex vivo experiments, and the assay was shown to be appropriate for clinical sample analysis. Myeloid cells exhibited a smaller proportion of acH4-positive cells than the lymphoid population, but a greater fold increase above basal levels.
CONCLUSIONS
CONCLUSIONS
The percentage of acH4-positive myeloid cells has the potential to be used as a robust pharmacodynamic biomarker for the measurement of acH4 for HDACIs. The developed assay is now being utilised in a clinical trial involving the treatment of childhood ependymoma patients with valproic acid.
Substances chimiques
Antineoplastic Agents
0
Histone Deacetylase Inhibitors
0
Histones
0
Valproic Acid
614OI1Z5WI
Types de publication
Clinical Trial, Phase II
Journal Article
Randomized Controlled Trial
Langues
eng
Sous-ensembles de citation
IM
Pagination
e13115Subventions
Organisme : Cancer Research UK
Organisme : Experimental Cancer Medicine Centre Network
Informations de copyright
© 2019 Stichting European Society for Clinical Investigation Journal Foundation.