MicroRNA­155 modulation of CD8+ T­cell activity personalizes response to disease­modifying therapies of patients with relapsing­remitting multiple sclerosis.

Multiple sclerosis (MS) is a chronic autoimmune disease where activated immune cells can attack oligodendrocytes causing damage to the myelin sheath. Several molecular mechanisms are responsible for the auto-activation of immune cells such as RNA interference (RNAi) through microRNAs (miRNAs or miRs). In the present study, the role of miR-155 in regulating CD8+ T-cell activity in patients with relapsing-remitting multiple sclerosis (RRMS) was investigated, in terms of its migratory functions with regard to intracellular adhesion molecule-1 (ICAM1) and integrin subunit ß2 (ITGB2), and its cytotoxic proteins, perforin and granzyme B. Gene expression of miR-155, ICAM1, ITGB2, perforin and granzyme B was evaluated following epigenetic modulations using reverse transcription-quantitative polymerase chain reaction in CD8+ T-cells isolated from blood samples of patients with RRMS and compared to healthy controls. The ectopic expression of miR-155 resulted in a persistent downregulation in all genes of interest related to CD8+ T-cell activation that were positively correlated with the Expanded Disability Status Scale of patients. The present study revealed the interplay between miR-155, ICAM1, and ITGB2, shedding light on their beneficial use as possible therapeutic regulators and diagnostic biomarkers of disease. Moreover, epigenetic modulations enhancing the efficacy of disease-modifying therapies (DMTs) may be employed as personalized therapy, to decrease the side effects of DMTs and improve the outcomes of patients.

Association between circulating MicroRNAs (hsa-miR-92a-1* and hsa-miR-454) and multiple sclerosis phenotypes and activity status.

Efficient diagnosis of multiple sclerosis (MS) disease along with early prediction of its progression will ultimately lead to better management, control of complications and improvement of therapeutic outcomes and patient's well-being. Blood based biomarkers like circulating microRNAs represent a non- invasive, fast, and easily measured markers with a promising potential. This work intended to assess the relative expression of circulating hsa-miR-454 and hsa-miR-92a-1* as a diagnostic and prognostic tool among Egyptian MS patients in terms of correlation to disease type and severity. hsa-miR-454 and hsa-miR-92a-1* relative expression was measured in the plasma of 31 MS patients, relapsing remitting MS (RRMS, n=21) and progressive MS (PMS, n=10) and 20 age and sex matched normal controls by using reverse transcription followed by real time PCR. Disease severity assessment was done in the form of patient expanded disability status scale (EDSS) evaluation. Relative expression of hsa-miR-454 and hsa-miR-92a-1* did not show a statistically significant difference between MS cases and controls. However, hsa-miR-454 was significantly higher among RRMS patients in comparison to PMS patients (P = 0.04). Additionally, both markers showed a statistically significant upregulation among patients in disease exacerbation in comparison to patients in remission (P = < 0.01) and both showed a negative correlation with EDSS. In conclusion, microRNAs may represent potential valuable non-invasive biomarkers for assessment of MS type (RRMS vs PMS), as well as for prediction of disease activity and severity in MS patients.

The MuSC-19 study: The Egyptian cohort.

OBJECTIVE: This study aimed to report the severity of COVID-19 in a cohort of Egyptian patients with multiple sclerosis (MS) with particular attention on the impact of disease modifying drugs (DMDs). METHODS AND STUDY POPULATION: We included 119 MS patients recruited from two centers, Ain-Shams university and Cairo university with confirmed or suspected COVID-19 during the period from May to September 2020 as a part of the MuSC-19 project. Univariate logistic regression was fitted to assess risk factors for severe COVID-19 (at least one outcome among hospitalization, ICU admission and death). RESULTS: Females were 77%, mean age was 34 years, mean duration of MS was 5.28 years, median EDSS was 3, most of the patients (83%) had RRMS, while 15% and 2% had respectively SPMS and PPMS. Only eleven patients (9% of study population) had a severe outcome and 3 patients (3%) died. Headache was the only symptom significantly associated with the severity of COVID-19 (OR=10.85, P = 0.001). There was no association between any of the DMDs and severe COVID-19 outcome. CONCLUSION: This study showed an acceptable safety profile of DMDs in Egyptian MS patients who developed COVID-19, as 91% of the cohort had a favorable outcome. Headache as a symptom associated with severe outcome in Egyptian patients' needs further validation.

miR-155 and functional proteins of CD8+ T cells as potential prognostic biomarkers for relapsing-remitting multiple sclerosis.

BACKGROUND: Multiple Sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) that results in neurological deficits in patients leading to disabilities which are evaluated on a scale known as the Expanded Disability Status Scale (EDSS). The most prevalent subtype of the disease is Relapsing-Remitting Multiple sclerosis (RRMS). One of the key players in MS pathogenesis is CD8+ T cells present in abundance in MS lesions expressing surface receptors, intracellular adhesion molecule (ICAM1) and integrin Subunit Beta 2 (ITGB2). These proteins are crucial for migration through the blood-brain barrier (BBB) and secondary stimulatory signal, along with the cytotoxic proteins perforin and granzymeB that attack oligodendrocytes. MicroRNAs (miRNAs) are small non-coding RNAs that play a substantial regulatory role in various disease pathogeneses through post-transcriptional modifications, and miR-155 shows potential for its use as a biomarker of the disease. The study aims at investigating the expression of miR-155, ICAM1, ITGB2, perforin and GranzymeB in CD8+ T cells of RRMS patients receiving different treatment regimens and how these genes correlate with patients' EDSS and miR-155 expression. METHODS: Gene expression of miR-155, ICAM1, ITGB2, perforin and granzymeB was evaluated using RT-qPCR in CD8+ T cells isolated from blood samples of RRMS patients and compared to healthy controls. RESULTS: Results showed downregulation of miR-155 and upregulation of surface receptors and cytotoxic proteins in CD8+T cells with significant correlation with each other and patients' EDSS. CONCLUSION: This study helps pave the road for the discussed genes for their use as potential biomarkers of disease disability and future investigations on their regulatory roles in disease pathogenesis.

Telephone-based assessment of multiple sclerosis patients at Ain Shams University Hospital in the coronavirus disease 2019 pandemic.

BACKGROUND: Assessment of multiple sclerosis (MS) patients during the era of the coronavirus disease 2019 (COVID-19) pandemic was confronted with the overwhelmed healthcare facilities in Egypt and fear of the patients to get infected while attending the follow-up visits. This study aimed to assess the value of telephone-based assessments in the follow-up of MS patients. It includes one hundred and five patients who participated in the study and completed 3 telephone-based assessments which are the Hauser Ambulation index, Multiple Sclerosis Neuropsychology Questionnaire (MSNQ), and Symptoms of Multiple Sclerosis Scale (SMSS). RESULTS: The Hauser Ambulation index was significantly correlated with the latest Expanded Disability Status Scale (EDSS) score done within 1 month from the telephone call (r=0.738, P<0.001). The analysis of MSNQ scores showed that one-third of the study population had evidence of cognitive and/or neuropsychological impairment. Post hoc analysis regarding the cognitive and psychological impairment component of SMSS revealed that the patients who answered "Never" had significantly lower MSNQ scores compared to those who answered "Sometimes" (P=0.016), "Often" (P=0.022), and "Always" (P=0.001). The comparison of the EDSS scores of the patients regarding the sensory-motor impairment component of SMSS showed a non-significant difference. CONCLUSION: The Hauser Ambulation index may be a reliable telephone-based tool for the assessment of physical disability. The MSNQ and the cognitive and psychological impairment component of SMSS can be used for the assessment of cognitive and psychological impairment among patients with MS.

Magnetic resonance imaging markers of disability in Egyptian multiple sclerosis patients.

BACKGROUND: The aim of this work was to identify the magnetic resonance imaging (MRI) markers of disability in Egyptian multiple sclerosis (MS) patients. SUBJECTS AND METHODS: This retrospective observational study included 673 patients recruited from the registry of the MS unit at Ain Shams University hospitals. At the time when the MRI scans of the brain and spinal cord were done (with and without gadolinium enhancement), clinical disability was rated using the Expanded Disability Status Scale (EDSS) during the patient's first visit. RESULTS: Females represented 72.5%, all types of MS were included, the mean age of onset was 26.1 ±â€¯7.7(SD) years, mean duration of illness was 8.3 ±â€¯5.5(SD) years. The mean EDSS of the patients was 3.5 ±â€¯2.1. The study population was divided into three groups according to the EDSS score; mild from 0-3 (56.6%), moderate from 3.5-6 (34.9%) and severe more than 6 (8.5%). The number and types of MRI lesions (T2, T1 black holes, T1 contrast and confluent lesions) in the different anatomical locations (periventricular, juxtacortical, infratentorial and spinal) were correlated with the clinical and demographic data of the patients as well as with the EDSS score. The presence of confluent brain lesions (P Ë‚ 0.001), brain T1 hypointense lesions (P = 0.009), and infratentorial T2 lesions (from 1 to 3 lesions (P = 0.04), from 4 to 10 (P ˂ 0.001) and more than 10 (P ˂ 0.001)), were significantly correlated to high EDSS scores after linear regression analysis. CONCLUSION: This is the first Egyptian study to show that infratentorial lesions, confluent brain lesions and T1 hypointense lesions are conventional MRI parameters that correlate with the degree of disability in Egyptian MS patients.

Vitamin D and body mass index in Egyptian multiple sclerosis patients.

BACKGROUND: Vitamin D deficiency and obesity may be related to the pathogenesis and disease activity of multiple sclerosis (MS). This study aimed to assess the correlation between the serum level of 25(OH) vitamin D, body mass index (BMI) and the Expanded Disability Status Scale (EDSS) in a sample of Egyptian MS patients. SUBJECTS AND METHODS: This was an observational study that included 130 MS patients who were recruited consecutively among the patients attending the MS unit of Ain Shams University Hospital, Cairo, in the period between December 2017 and March 2018. The serum level of 25(OH) D, BMI and EDSS were recorded. RESULTS: Females represented 77.7% of the study sample, the mean age was 32.4 ±â€¯8.2 years. MS types were: RRMS 83.1%, SPMS 14.6% and PPMS in 2.3%. Serum level of 25(OH) vitamin D was deficient (less than 10 ng/ml) in 69.2% and insufficient (10-30 ng/ml) in 19.2% of the study population. The mean BMI was 25.5 ±â€¯4.7 kg/m² (classified as overweight). The mean EDSS was 3.5 ±â€¯1.9. The relationship between the EDSS score and 25(OH) D level was inversely correlated, all patients with EDSS ≤ 2 had sufficient levels while all patients with EDSS ≥ 4.5 had deficient levels. High EDSS scores were statistically correlated (p < 0.001) to high BMI and low Log 25(OH) D levels. An inverse correlation was found between the BMI and log 25(OH) D. CONCLUSION: Vitamin D deficiency and overweight are predominant among Egyptian MS patients. The EDSS was positively correlated to the BMI and negatively correlated to 25(OH) D. These factors may possibly play a role in the pathogeneses and progression of MS in Egypt.

MicroRNA 26a Expression in Peripheral Blood Mononuclear Cells and Correlation with Serum Interleukin-17 in Relapsing-Remitting Multiple Sclerosis Patients.

Multiple sclerosis (MS) is a chronic multifocal inflammatory demyelinating disease. One of the main cells that play a crucial role in pathogenesis of MS is T helper 17 (Th 17). There are growing interests in nominating microRNAs in Th17 cell differentiation and suggesting new therapeutic modalities. The aim of the study was to assess microRNA 26a (miR26a) expression in peripheral blood mononuclear cells of relapsing - remitting MS patients as compared to healthy control subjects and examine association of these levels with serum IL17. Forty (40) relapsing - remitting MS patients were enrolled based on the MacDonald criteria (20 in relapsing phase and 20 in remitting phase). In addition, twenty (20) healthy control subjects were included. Blood samples were subjected to quantitative polymerase chain reaction (qPCR) for miR26a and ELISA for serum IL 17 levels. A significant upregulation of miR26a relative expression level (∆∆ Ct) and serum IL17 level (pg/ml) was found in total MS patients and remitting MS patients when compared with controls (P < 0.001). Among the relapsing group, a significant increase in miR26a expression levels (P= 0.004) but not serum IL17 level was demonstrated. Insignificant correlation between miR26a expression and serum IL17 in MS patients was detected (r= 0.08, P= 0.62). In conclusion, a significant increase of these two biomarkers (miR26a & IL17) occurs in relapsing - remitting MS patients, and this reflects their important role in pathogenesis and disease development.

Clinical characteristics of patients with multiple sclerosis enrolled in a new registry in Egypt.

BACKGROUND: Epidemiological studies of multiple sclerosis (MS) are lacking in Egypt. OBJECTIVE: To study the characteristics of Egyptian patients with multiple sclerosis in a new registry in a major tertiary referral centre in Cairo, Egypt. SUBJECT AND METHODS: Patients were from the project MS database of the Multiple Sclerosis Unit at Ain Shams University Hospitals (N=950). We conducted a detailed medical history and examination including the Expanded Disability Status Scale (EDSS). RESULTS: Females represented 72% of subjects (female: male ratio 2.57:1). The mean age of disease onset was 26.1±7.6 years. Relapsing-remitting MS (RRMS) was the most common presentation (74.6%). Visual or sensory symptoms were the most common at presentation with RRMS, while motor symptoms were the most common presentation in other types of MS. Time to diagnosis was delayed up to 2 years in 27.8% of patients. The mean EDSS score was 3.6±2.1; 55% had EDSS≤3. About half (49%) received a disease-modifying drug. Progressive MS and motor presentation were associated with higher disability. CONCLUSIONS: This is the first documented MS registry from Egypt. The clinical characteristics of MS in Egypt was similar to other Arab countries and western countries. MS is more common among females in Egypt, with RRMS being the most common presentation. Visual symptoms and motor symptoms were the most common presentations in RRMS and progressive MS, respectively. Our findings also highlight the value of establishing registries in Egypt in order to be able to study, prospectively, the clinical course of the disease, the response to various DMD's and the epidemiology of MS in Egypt.