Cis-vaccenic acid induces differentiation and up-regulates gamma globin synthesis in K562, JK1 and transgenic mice erythroid progenitor stem cells

Aimola, Idowu A.; Inuwa, Hajiya Mairo; Nok, Andrew J.; Mamman, Aisha Indo; Bieker, James J.

doi:10.1016/j.ejphar.2016.02.041

Cited by 5 publications

(4 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Overall, the K562 cell experimental repeats were similar and although there was expected inter-individual variation in the degree and timing of up- and/or down-regulation of γ-globin and its key regulators in the erythroid model, the trends remained true and largely comparable. Despite the fact that several studies have and continue to utilise K562 as a model to investigate various components of γ-globin expression [ 39 , 53 , 74 – 77 ], it could be argued that K562 cells are not an ideal model for studying globin switching because of their bias expression of γ-globin. We however, saw this as an advantage in a number of ways: (1) the aim of this study was not to demonstrate the switching in expression of the globin genes (β to γ) but rather to use a stable cell line model in conjunction with cord blood-derived erythroid cells to demonstrate increases in γ-globin and decreases its regulators ( BCL11A, KLF - 1 and MYB ) in response to HU, and for this purpose, K562 cells would suffice as a model; (2) in choosing the whether to use miRNA mimics or anti-miRNAs (miR-inhibitors), the high expression of γ-globin in K562 was considered; if our hypothesis of HU-induced miRNA-mediated repression of MYB was correct, the use of mimics would not only introduce unnaturally high levels of miRNAs into this model (which we felt may in fact result in some artefactual results merely because of this unnatural state of aberrant concentration of miRNAs in the cells) but it would also be compounded by any co-treatments with HU in causing γ-globin expression.…”

Section: Discussionmentioning

confidence: 99%

Hydroxyurea down‐regulates BCL11A, KLF‐1 and MYB through miRNA‐mediated actions to induce γ‐globin expression: implications for new therapeutic approaches of sickle cell disease

Pule

Mowla

Novitzky

et al. 2016

Clinical & Translational Med

View full text Add to dashboard Cite

Background The major therapeutic benefit of hydroxyurea, the only FDA-approved pharmacologic treatment for sickle cell disease (SCD), is directly related to fetal hemoglobin (HbF) production that leads to significant reduction of morbidity and mortality. However, potential adverse effects such as infertility, susceptibility to infections, or teratogenic effect have been subject of concerns. Therefore, understanding HU molecular mechanisms of action, could lead to alternative therapeutic agents to increase HbF with less toxicity. This paper investigated whether HU-induced HbF could operate through post-transcriptional miRNAs regulation of BCL11A, KLF - 1 and MYB, potent negative regulators of HbF. Both ex vivo differentiated primary erythroid cells from seven unrelated individuals, and K562 cells were treated with hydroxyurea (100 μM) and changes in BCL11A , KLF - 1 , GATA - 1 , MYB, β- and γ-globin gene expression were investigated. To explore potential mechanisms of post-transcriptional regulation, changes in expression of seven targeted miRNAs, previously associated with basal γ-globin expression were examined using miScript primer assays. In addition, K562 cells were transfected with miScript miRNA inhibitors/anti-miRNAs followed by Western Blot analysis to assess the effect on HbF protein levels. Direct interaction between miRNAs and the MYB 3′-untranslated region (UTR) was also investigated by a dual-luciferase reporter assays. Results Down-regulation of BCL11A and MYB was associated with a sevenfold increase in γ-globin expression in both primary and K562 cells (p < 0.003). Similarly, KLF - 1 was down-regulated in both cell models, corresponding to the repressed expression of BCL11A and β-globin gene (p < 0.04). HU induced differential expression of all miRNAs in both cell models, particularly miR-15a, miR-16, miR-26b and miR-151-3p. An HU-induced miRNAs-mediated mechanism of HbF regulation was illustrated with the inhibition of miR-26b and -151-3p resulting in reduced HbF protein levels. There was direct interaction between miR-26b with the MYB 3′-untranslated region (UTR). Conclusions These experiments have shown the association between critical regulators of γ-globin expression ( MYB , BCL11A and KLF - 1 ) and specific miRNAs; in response to HU, and demonstrated a mechanism of HbF production through HU-induced miRNAs inhibition of MYB . The role of miRNAs-mediated post-tra...

show abstract

Section: Discussionmentioning

confidence: 99%

Hydroxyurea down‐regulates BCL11A, KLF‐1 and MYB through miRNA‐mediated actions to induce γ‐globin expression: implications for new therapeutic approaches of sickle cell disease

Pule

Mowla

Novitzky

et al. 2016

Clinical & Translational Med

View full text Add to dashboard Cite

show abstract

“…Cisvaccenic acid is a monounsaturated n -7 fatty acid and is reported to have antibacterial and hypolipidemic effects in rats (Hamazakiet al, 2016). Cis-vaccenic acid may have multiple modes of activity such as antioxidant activity which involves the neutralization of free radicals hence reducing oxidative stress, it may serve as an antiinflammatory agent by decreasing the production of cytokines that promote inflammation; it may also act as a transcription factor regulating the expression of genes that are involved in metabolic processes (Aimola et al, 2016., Rontani et al, 2003. Hexadecanoic acid methyl ester was reported to have bactericidal effect against multi drug resistant bacteria (Shaaban et al, 2021).…”

Section: Resultsmentioning

confidence: 99%

Identification of Bioactive Compounds in Chrysobalanus icaco Seed Kernel Using Gas Chromatography-Mass Spectrometry

N. C.,

T.,

S. O.

2024

Am. J. Food Sci. Technol.

View full text Add to dashboard Cite

Chrysobalanus icaco seed kernel (i.e Cocoplum) is good source of oil and is utilized as spice in soups or stews in some African cuisines. The work aimed to identify compounds in methanol and dichloromethane/methanol (1:1, v/v) flour extract from its seed kernels using gas chromatography-mass spectrometry. Eleven compounds were identified from the methanol extract while thirteen compounds were identified from the dichloromethane/methanol extract. The most abundant compounds from the methanol extract were cis-13-octadecenoic acid methyl ester, cis vaccenic acid and n-Hexadecanoic acid. They had relative abundance of 28.83, 17.14 and 15.40% respectively. 2,6-octadienal-3,7-dimethyl (E) (ie α-citral), citral, cis-3-hexenyl cis-3-hexenoate and trans-2,7-dimethyl-4,6-octadiene-2-ol were the most abundant in the dichloromethane/methanol extract. These had relative abundance of 12.86, 18.92, 10.56 and 20.59% respectively. Compounds which were common to both methanol and dichloromethane/methanol extracts of C. icaco seed kernel were 2,6-octadienal-3,7-dimethyl (E), hexadecanoic acid methyl ester, n-Hexadecanoic acid, heptadecanoic acid, 16 methyl, methyl ester, cis-vaccenic acid and oleic acid. Both solvents extracted varied concentrations of compounds which fall in different classes of either being an unsaturated long chain aldehyde, fatty acids, fatty acid methyl esters, indene derivatives, monoterpenoid alchohol. The compounds identified in both extracts of Chrysobalamus icaco seed kernels have various beneficial bioactivities and sensory attributessuch as being flavor compounds which confers C.icaco seed kernel a peculiar aroma. As such, Chrysobalamus icaco seed kernel is a functional food and can serve as a good raw material for edible oil from which bioactive compounds can be isolated and utilized in relevant food systems and in the preparation of neutraceuticals and pharmaceuticals. Therefore, its use in cuisines is greatly encouraged.

show abstract

“…RNA samples were subjected to cDNA synthesis for 1 h at 42°C via a RevertAid first-strand cDNA synthesis kit (Thermo Scientific Baltics, UAB). The obtained cDNA was used as a template, and qRT-PCR was performed using a Maxima SYBR Green/ROX qPCR Master Mix (2X) (K0221, Thermo Scientific Baltics, UAB) on an Mx3000P QPCR System (Agilent Technologies, United States) as previously described conditions ( Aimola et al, 2016 ). The expression of the gene was evaluated using the 2 −ΔΔCt method and normalized by using the reference GAPDH gene.…”

Section: Methodsmentioning

confidence: 99%

In Vitro and In Vivo Studies for the Investigation of γ-Globin Gene Induction by Adhatoda vasica: A Pre-Clinical Study of HbF Inducers for β-Thalassemia

Iftikhar¹,

Rahman

Khan³

et al. 2022

Front. Pharmacol.

View full text Add to dashboard Cite

Fetal hemoglobin (HbF) is a potent genetic modifier, and the γ-globin gene induction has proven to be a sustainable therapeutic approach for the management of β-thalassemia. In this study, we have evaluated the HbF induction ability of A. vasica in vitro and in vivo, and the identification of potential therapeutic compounds through a bioassay-guided approach. In vitro benzidine-Hb assay demonstrated strong erythroid differentiation of K562 cells by A. vasica extracts. Subsequently, an in vivo study with an aqueous extract of A. vasica (100 mg/kg) showed significant induction of the γ-globin gene and HbF production. While in the acute study, the hematological and biochemical indices were found to be unaltered at the lower dose of A. vasica. Following the bioassay-guided approach, two isolated compounds, vasicinol (1) and vasicine (2) strongly enhanced HbF levels and showed prominent cellular growth kinetics with ample accumulation of total hemoglobin in K562 cultures. High HbF levels were examined by immunofluorescence and flow cytometry analysis, concomitant with the overexpression in the γ-globin gene level. Compound 1 (0.1 µM) and compound 2 (1 µM) resulted in a greater increase in F-cells (90 and 83%) with marked up (8-fold and 5.1-fold) expression of the γ-globin gene, respectively. Molecular docking studies indicated strong binding affinities of (1) and (2) with HDAC2 and KDM1 protein that predict the possible mechanism of compounds in inhibition of these epigenetic regulators in the γ-globin gene reactivation. Altogether, these observations demonstrated the therapeutic usefulness of A. vasica for fostering HbF production in clinical implications for blood disorders.

show abstract

Cis-vaccenic acid induces differentiation and up-regulates gamma globin synthesis in K562, JK1 and transgenic mice erythroid progenitor stem cells

Cited by 5 publications

References 56 publications

Hydroxyurea down‐regulates BCL11A, KLF‐1 and MYB through miRNA‐mediated actions to induce γ‐globin expression: implications for new therapeutic approaches of sickle cell disease

Hydroxyurea down‐regulates BCL11A, KLF‐1 and MYB through miRNA‐mediated actions to induce γ‐globin expression: implications for new therapeutic approaches of sickle cell disease

Identification of Bioactive Compounds in Chrysobalanus icaco Seed Kernel Using Gas Chromatography-Mass Spectrometry

In Vitro and In Vivo Studies for the Investigation of γ-Globin Gene Induction by Adhatoda vasica: A Pre-Clinical Study of HbF Inducers for β-Thalassemia

Contact Info

Product

Resources

About