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

NCT ID: NCT05371743

Last Updated: 2023-04-12

Basic Information

• Recruitment Status: UNKNOWN
• Total Enrollment: 2 participants
• Study Classification: OBSERVATIONAL
• Study Start Date: 2022-05-01
• Study Completion Date: 2023-09-01

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

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.

Conditions

Primary Immune Thrombocytopenia

Study Design

• Observational Model Type: CASE_CONTROL
• Study Time Perspective: CROSS_SECTIONAL

Study Groups

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:

1. Secondary causes of ITP as systemic lupus erythematous (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.

Plasma RNA isolation, qPCR analysis of micro RNA

Intervention Type: DIAGNOSTIC_TEST

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

estimation of serum level of IL2

Intervention Type: DIAGNOSTIC_TEST

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

estimation of serum level of IL17

Intervention Type: DIAGNOSTIC_TEST

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.

No interventions assigned to this group

Interventions

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

Intervention Type: 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

Intervention Type: 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

Intervention Type: DIAGNOSTIC_TEST

Eligibility Criteria

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.

• Minimum Eligible Age: 18 Years
• Maximum Eligible Age: 90 Years
• Eligible Sex: ALL
• Accepts Healthy Volunteers: Yes

Sponsors

Sohag University

OTHER

Sponsor Role: lead

Responsible Party

Noha Saber Shafik

lecturer of Medical Microbiology and Immunology, faculty of medicine

Responsibility Role: PRINCIPAL_INVESTIGATOR

Principal Investigators

Hend M Moness, professor

Role: PRINCIPAL_INVESTIGATOR

Faculty Of Medicine, Minya university

Aliaa S Abd EL Fatah, professor

Role: PRINCIPAL_INVESTIGATOR

Faculty Of Medicine, Minya university

Rasha F Ahmed, professor

Role: PRINCIPAL_INVESTIGATOR

Faculty of medicine, Minya university

Locations

Sohag University

Sohag, , Egypt

Site Status: RECRUITING

Countries

Egypt

Central Contacts

Noha S Shafik, lecturer

Role: CONTACT

Nohasaber@med.sohag.edu.eg

01067261504 ext. +20

Doaa M Elroby, lecturer

Role: CONTACT

dosa.elroby@pharm.sohag.edu.eg

01009374489 ext. +20

Facility Contacts

N S Shafik, lecture

Role: primary

nohasaber@med.sohag.edu.eg

01067261504 ext. 02

M M Abd El Rhman, lecturer

Role: backup

monamohamed@med.sohag.edu.eg

01021025859 ext. 02

References

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.

Reference Type: BACKGROUND

PMID: 24433387 (View on PubMed)

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.

Reference Type: BACKGROUND

PMID: 22537755 (View on PubMed)

Other Identifiers

290-2022

Identifier Type: -

Identifier Source: org_study_id