microRNAITP: Plasma microRNA Levels and Some Cytokines Expression in Patients With ITP Primary Immune Thrombocytopenic Purpura (ITP)
Study Details
Study Description
Brief Summary
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
| Condition or Disease | Intervention/Treatment | Phase |
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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 Design
Arms and Interventions
| Arm | 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: malignancy megaloblastic anemia aplastic anemia lymphoproliferative disorders liver disease renal impairment pregnancy 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
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| normal individuals Blood samples will be taken from normal individuals. |
Outcome Measures
Primary Outcome Measures
- Evaluation of some plasma miRNA profiling in primary ITP and correlation to disease phases and their possible roles in pathogenesis of ITP [1 May 2022 to 1 September]
Measurement of Micro RNA by qPCR
- 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 [1 May 2022 to 1 September]
Measurement by ELISA
- 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 [1 May 2022 to 1 September]
Measurement by ELISA
Eligibility Criteria
Criteria
Inclusion Criteria:
- ITP patients
Exclusion Criteria:
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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:
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malignancy
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megaloblastic anemia
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aplastic anemia
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lymphoproliferative disorders
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liver disease
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renal impairment
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pregnancy 3-Organomegally and/or lymphadenopathy. 4-Recent history of vaccination. 5-Recent evidence of bacterial infection.
Contacts and Locations
Locations
| Site | City | State | Country | Postal Code | |
|---|---|---|---|---|---|
| 1 | Sohag University | Sohag | Egypt | 82733 |
Sponsors and Collaborators
- 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
Study Documents (Full-Text)
None provided.More Information
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.
- 290-2022