Plasma microRNA Levels and Some Cytokines Expression in Patients With ITP Primary Immune Thrombocytopenic Purpura (ITP) (microRNAITP)

Plasma microRNA Levels and Some Cytokines Expression in Patients With Primary Immune Thrombocytopenic Purpura (ITP)

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by low platelet counts with or without mucocutaneous bleeding (McMillan, 2007). Like the majority of autoimmune diseases, ITP is an organ-specific disease, and abnormalities in the regulation of the immune system have been shown to play an important role in the initiation and/or perpetuation of the disease (McKenzie et al.,2013).

Still, immune thrombocytopenia (ITP) is a significant clinical problem due to chronicity, treatment cost, occurrence mainly in, young, and relatively poorer quality of life

Study Overview

• Status: Recruiting
• Conditions: Primary Immune Thrombocytopenia
• Intervention / Treatment: Diagnostic test: Plasma RNA isolation, qPCR analysis of micro RNA; Diagnostic test: estimation of serum level of IL2; Diagnostic test: estimation of serum level of IL17

Detailed Description

In recent years the critical role of miRNAs has been established in many diseases, including autoimmune disorders. Immune thrombocytopenic purpura (ITP) is a predominant autoimmune disease, in which aberrant expression of miRNAs has been observed, suggesting that miRNAs are involved in its development (Jafarzadeh et al., 2021). Studies have also shown that cell-free miRNAs in circulation are stable and that such miRNAs may be exploited as novel disease markers (Etheridge et al.,2011) and ( van Rooij et al., 2008).

MicroRNAs (miRNAs) are endogenous small RNAs, usually 18-25 nucleotides in length. These non-coding RNAs regulate gene expression by several mechanisms, such as repressing protein translation and altering mRNA stability (Ambros, 2008. Bartel,2004). In humans, >2,000 miRNAs have been discovered. The functional significance of the majority of the identified miRNAs has yet to be fully elucidated. Studies have shown that miRNAs play important roles in hematopoietic differentiation, e. g. megakaryocytopoiesis. (Garzon et al., 2008) and erythropoiesis ( Masaki et al., 2007 )and (Bruchovaetal., 2007), and in hematological malignancies (Rossi et al.,2010) and (Visone et al.,2009). More recently, miRNAs have also been implicated in cellular immune responses that contribute to ITP (Jernas et al., 2013) and (McKenzie etal., 2013). It was found that 23 differentially expressed miRNAs in ITP (14 up-regulated and 9 down-regulated) Altered miRNA expression may occur in specific diseases and at specific disease stages (Martin et al., 2012) Also, Recent studies have demonstrated that Th17, which is characterized for its production of IL-17, is elevated in ITP patients (Hu et al., 2012) and (Huber et al., 2007). IL-17 belongs to the IL-17 cytokine family. Increased IL-17 expression has been observed in various autoimmune diseases, such as rheumatoid arthritis (RA) ( Roeleveld et al., 2013) and systemic lupus erythematosus (SLE) (Ballantine et al., 2014). This evidence suggests that IL-17 may be associated with autoimmune diseases.

• Study Type: Observational
• Enrollment (Anticipated): 2

Contacts and Locations

• Study Contact: Name: Noha S Shafik, lecturer; Phone Number: +20 01067261504; Email: Nohasaber@med.sohag.edu.eg
• Study Contact Backup: Name: Doaa M Elroby, lecturer; Phone Number: +20 01009374489; Email: dosa.elroby@pharm.sohag.edu.eg

Study Locations

• Egypt
  • Sohag, Egypt, 82733
    • Recruiting
    • Sohag University
    • Contact: N S Shafik, lecture; Phone Number: 02 01067261504; Email: nohasaber@med.sohag.edu.eg
    • Contact: M M Abd El Rhman, lecturer; Phone Number: 02 01021025859; Email: monamohamed@med.sohag.edu.eg
    • Sub-Investigator: S B Hemdan, lecturer
    • Sub-Investigator: R M Farag, lecturer
    • Principal Investigator: A N Elsayed, lecturer

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 90 years (Adult, Older Adult)
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

This is a cross-sectional study. Patients will be recruited from the internal department- hematology unit outpatient clinic of El Minia University Hospital in collaboration with the clinical pathology department El Minia University Hospital and the biochemistry department of Minia and Sohag University. Research Ethics Medical Review Board of El-Minia University has approved the protocol.

Description

Inclusion Criteria:

• ITP patients

Exclusion Criteria:

• 1- Secondary causes of ITP as systemic lupus erythematosus (SLE), viral infections (HIV, hepatitis B or C infections) 2- Other underlying medical diseases that may cause thrombocytopenia as:
• malignancy
• megaloblastic anemia
• aplastic anemia
• lymphoproliferative disorders
• liver disease
• renal impairment
• pregnancy 3-Organomegally and/or lymphadenopathy. 4-Recent history of vaccination. 5-Recent evidence of bacterial infection.

Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
ITP patients; Patients will be recruited from the internal department- hematology unit outpatient clinic of El Minia University Hospital in collaboration with the clinical pathology department of El Minia University Hospital and the biochemistry department of Minia and Sohag University. Exclusion criteria: Secondary causes of ITP as systemic lupus erythematous (SLE), viral infections (HIV, hepatitis B or C infections); Other underlying medical diseases that may cause thrombocytopenia as:malignancymegaloblastic anemiaaplastic anemialymphoproliferative disordersliver diseaserenal impairmentpregnancy; Organomegally and/or lymphadenopathy.; Recent history of vaccination.; Recent evidence of bacterial infection.	Diagnostic test: Plasma RNA isolation, qPCR analysis of micro RNA; qPCR will be performed using Taqman microRNA assays (Applied Biosystems, Foster City, CA, USA) according to the manufacturer's protocol. Total RNAs going to be used to make cDNAs using TaqMan microRNA RT Kit. Diluted cDNAs were mixed with TaqMan Universal PCR Master Mix (No AmpErase UNG). Taqman miRNA assay will run in the 7500 Real-Time PCR System (Applied Biosystems). miR-39 also will be used as an exogenous control. All assays will be done in triplicates. The expression levels will be evaluated using the comparative cycle threshold (∆∆_Ct) method; Diagnostic test: estimation of serum level of IL2; 2ml of patient serum will be used to measure IL 2 in patients with different groups by ELISA technique. The techniques will be done in the central research laboratory in Sohag university hospital; Diagnostic test: estimation of serum level of IL17; 2ml of patient serum will be used to measure IL17 in patients with different groups by ELISA technique. The techniques will be done in the central research laboratory in Sohag university hospital
normal individuals; Blood samples will be taken from normal individuals.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Evaluation of some plasma miRNA profiling in primary ITP and correlation to disease phases and their possible roles in pathogenesis of ITP	Measurement of Micro RNA by qPCR	1 May 2022 to 1 September
Explore the cytokines level production of IL-2 in patients with primary ITP at different disease phases and its possible role in pathogenesis of primary ITP	Measurement by ELISA	1 May 2022 to 1 September
Explore the cytokines level production of IL-17 in patients with primary ITP at different disease phases and its possible role in pathogenesis of primary ITP	Measurement by ELISA	1 May 2022 to 1 September

Collaborators and Investigators

• Sponsor: Sohag University
• Investigators: Principal Investigator: Hend M Moness, professor, Faculty of medicine, Minya university; Principal Investigator: Aliaa S Abd EL Fatah, professor, Faculty of medicine, Minya university; Principal Investigator: Rasha F Ahmed, professor, Faculty of medicine, Minya university

Publications and helpful links

General Publications

• Ballantine LE, Ong J, Midgley A, Watson L, Flanagan BF, Beresford MW. The pro-inflammatory potential of T cells in juvenile-onset systemic lupus erythematosus. Pediatr Rheumatol Online J. 2014 Jan 16;12:4. doi: 10.1186/1546-0096-12-4.
• Hu Y, Li H, Zhang L, Shan B, Xu X, Li H, Liu X, Xu S, Yu S, Ma D, Peng J, Hou M. Elevated profiles of Th22 cells and correlations with Th17 cells in patients with immune thrombocytopenia. Hum Immunol. 2012 Jun;73(6):629-35. doi: 10.1016/j.humimm.2012.04.015. Epub 2012 Apr 23.

Study record dates

Study Major Dates

• Study Start (Actual): May 1, 2022
• Primary Completion (Actual): September 1, 2022
• Study Completion (Anticipated): September 1, 2023

Study Registration Dates

• First Submitted: May 7, 2022
• First Submitted That Met QC Criteria: May 7, 2022
• First Posted (Actual): May 12, 2022

Study Record Updates

• Last Update Posted (Actual): April 12, 2023
• Last Update Submitted That Met QC Criteria: April 11, 2023
• Last Verified: April 1, 2023

More Information

Terms related to this study

• Keywords: MicroRNA, ITP, IL17
• Additional Relevant MeSH Terms: Pathologic Processes; Immune System Diseases; Autoimmune Diseases; Hematologic Diseases; Hemorrhage; Hemorrhagic Disorders; Blood Coagulation Disorders; Skin Manifestations; Blood Platelet Disorders; Thrombotic Microangiopathies; Purpura; Purpura, Thrombocytopenic; Purpura, Thrombocytopenic, Idiopathic; Thrombocytopenia
• Other Study ID Numbers: 290-2022

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No