Effect of DNMT SNPs on DNA Methylation in Primary ITP

Effect of DNA Methyltransferase 3A-448 G/A and 3B-149 C/T Single Nucleotide Polymorphisms on Global DNA Methylation in Primary Immune Thrombocytopenia

Sponsors

Lead Sponsor: Assiut University

Source Assiut University
Brief Summary

- To compare global DNA methylation status between ITP patients and healthy subjects .

- To determine the effect of DNMT3A -448 G/A SNP variant A allele and DNMT3B -149C/T SNP variant T-allele on global DNA methylation in both ITP patients and healthy control subjects.

Detailed Description

Immune thrombocytopenia (ITP), is an immune-mediated acquired disease of adults and children characterized by transient or persistent decrease of the platelet count and, depending upon the degree of thrombocytopenia, increased risk of bleeding (Rodeghiero et al., 2009). International guidelines define thrombocytopenia as a peripheral blood platelet count less than 100 000/uL (Raj, 2017). Platelet count values between 100 000/uL and 150 000/uL are frequently found in apparently healthy people (Adibi et al., 2007) and this threshold reduces the concern over the mild "physiologic" thrombocytopenia associated with pregnancy (Neunert et al., 2011).

Immune thrombocytopenia may be primary or secondary. Primary ITP is a diagnosis of exclusion. Secondary ITP occur due to an underlying diseases/conditions. Causes of secondary ITP include infection (CMV, Helicobacter pylori, HCV, HIV, Varicella zoster), Systemic lupus erythematosus, Antiphospholipid syndrome, Drug-induced, Lymphoproliferative disorders, Post bone marrow transplantation, and Post vaccination (Neunert et al., 2011).

Primary ITP is an acquired autoimmune disease causing both increased platelet destruction and insufficient platelet production. More than one mechanism could contribute to this including autoreactive B lymphocytes, Th1/Tc1 polarization, T-cell-mediated platelet lysis and abnormal circulating Treg cells. Also, DNA methylation may participate in the pathophysiology of ITP (Semple et al., 2010, Wang et al., 2011).

DNA methylation is a heritable, stable, and also reversible way of DNA modification; it can regulate gene expression without changing the nucleotide sequences (Huiyuan et al., 2013). DNA methylation is mediated by DNA methyltransferases (DNMTs). There are five members in DNMT group including DNMT3A and DNMT3B (Okano et al., 1999). DNMT3A and DNMT3B catalyze de novo methylation and it is important in the establishment of DNA methylation patterns within the embryo and during fetal development (Sawalha, 2008).

DNA methylation status takes part in the regulation of immune response, the loss of methylation pattern in immune cells will result in autoimmune disease by inducing aberrant gene expression. ITP is an autoimmune disease with many immune deficiencies with abnormal DNA methylation involved in the disease etiology (Huiyuan et al., 2013). Chen et al., 2011, concluded that aberrant DNA methylation may take part in the pathogenesis of ITP by quantifying the methylcytosine concentration of genomic DNA. They found hypomethylation pattern in CD4 T cells of ITP patients.

DNMT3A, and DNMT3B DNA methyltransferases are encoded by different genes on distinct chromosomes (Sawalha, 2008). There are many single nucleotide polymorphisms (SNPs) in DNMT3A gene which may affect catalytic activity of the DNMT3A enzyme, including -448G/A SNP (Zhao et al., 2012). Twenty-one polymorphisms have been identified in the DNMT3B gene including -149 C/T SNP and -579 G/T SNP (Zhang et al., 2015).

In a previous study in Assiut University Hospital (AbdelKader et al., 2018), the DNMT3A -448 G/A SNP variant A allele was significantly associated with decreased risk of primary ITP, while, DNMT3B -149C/T SNP variant T-allele was significantly associated with nearly double-fold increase in risk of primary ITP. However, the underlying mechanism behind this association was not investigated. Authors recommended to study mRNA expression of DNMT genes, enzymatic activity of DNA methyltransferases, quantification of global methylated DNA, and/or methylation status of methylation-sensitive genes involved in primary ITP pathogenesis to understand this association.

Overall Status Not yet recruiting
Start Date December 1, 2019
Completion Date December 1, 2021
Primary Completion Date October 1, 2021
Study Type Observational
Primary Outcome
Measure Time Frame
Comparison of global DNA methylation status between ITP patients and healthy subjects . baseline .
Enrollment 90
Condition
Intervention

Intervention Type: Genetic

Intervention Name: Global DNA methylation will be quantified in the DNA samples by colorimetry

Description: Global DNA methylation will be quantified in the DNA samples by colorimetry

Eligibility

Sampling Method: Non-Probability Sample

Criteria:

Inclusion Criteria:

- . Egyptian patients with isolated thrombocytopenia, no organomegaly or lymphoadenopathy, no constitutional symptoms (bone pains, weight loss, and night sweats) and no history of preceding drug intake (quinine, heparin) .

Exclusion Criteria:

- Conditions/diseases associated with secondary ITP.

Gender: All

Minimum Age: 18 Years

Maximum Age: 75 Years

Healthy Volunteers: Accepts Healthy Volunteers

Overall Official
Last Name Role Affiliation
Heba A Abdel-Hafeez, Professor Study Director Clinical Pathology Department in Assuit University Hospital
Overall Contact

Last Name: Mennat-Allah A Mahmoud, physician

Phone: 01006044750

Email: [email protected]

Verification Date

September 2019

Responsible Party

Type: Principal Investigator

Investigator Affiliation: Assiut University

Investigator Full Name: Mennat-Allah Abdelnasser Mahmoud Ahmed

Investigator Title: Clinical Pathology Department

Has Expanded Access No
Condition Browse
Arm Group

Label: primary ITP patients .

Description: 60 primary ITP patients .

Label: healthy subjects .

Description: 30 healthy age and sex matched control subjects .

Study Design Info

Observational Model: Case-Control

Time Perspective: Retrospective

Source: ClinicalTrials.gov