the EXPOSITION Study (EXPOSITION)

EXPOSITION (EXPosome, Oxidative Stress and InflammaTION) in Persons With Multiple Sclerosis Study

This is a cross-sectional study to evaluate the variation of biological biomarkers of oxidative stress and inflammation in response to the external exposome, in people with Multiple Sclerosis (pwMS).The objective is to study the variation of biological biomarkers of oxidative stress and inflammation in response to external exposome in pwMS, controlling for other biomarkers (cytokine, neurofilaments, microbiome), gender, age, anthropometric measurements, vitamin D levels and medical history. Specifically, the variation of microRNAs is defined as the primary outcome, in response to urban air pollution, urbanization, lifestyle and quality of life components of the external exposome. Following the functional exposome approach:(1)Information on a pwMS sample about socio-demographic characteristics and medical history will be collected and specific components of the (2) On the same pwMS sample, the internal exposome variation will be measured. MicroRNA levels and gut and nasal microbiota alpha- and beta-diversity and relative bacterial abundances will be considered as biomarkers of oxidative stress and inflammation. At the same time, cytokines and neurofilament proteins (NfL) will be measured as biomarkers of neurodegeneration and axonal damage. Adults (≥ 18 years) pwMS, with relapsing-remitting course, diagnosis of MS according to 2017 McDonald criteria and residing in Pavia or Milan (Italy) will be included. Potentially eligible pwMS will be screened by a neurologist expert in MS who will verify that all the inclusion criteria will be fulfilled. To validate variation among 7 selected MS diagnostic miRNA, in response to urban air pollution, urbanization, lifestyle and quality of life components of the external exposome, the differential expression (ΔCT) for each miRNA will be considered as the outcome measure. Two hundred eligible pwMS who meet the inclusion criteria and sign the informed consent will be included in the study, to consider 15% dropout at the blood sampling stage.

Study Overview

• Status: Not yet recruiting
• Conditions: Multiple Sclerosis, Relapsing-Remitting; Environmental Exposure; Lifestyle Factors

• Study Type: Observational
• Enrollment (Estimated): 200

Contacts and Locations

• Study Contact: Name: Eleonora Tavazzi; Phone Number: +39 0382380403; Email: eleonora.tavazzi@mondino.it
• Study Contact Backup: Name: Hellas Cena; Phone Number: +39 3357699199; Email: hellas.cena@unipv.it

Study Locations

• Italy
  • Pavia, Italy, 27100
    • IRCCS Foundation C. Mondino
    • Contact: Eleonora Tavazzi; Phone Number: +39 0382 380403; Email: eleonora.tavazzi@mondino.it
    • Sub-Investigator: Roberto Bergamaschi
    • Principal Investigator: Eleonora Tavazzi
    • Sub-Investigator: Hellas Cena
    • Sub-Investigator: Rachele de Giuseppe
    • Sub-Investigator: Beatrice Maccarini
    • Sub-Investigator: Aliki Kalmpourtzidou
    • Sub-Investigator: Elena Colombo
    • Sub-Investigator: Eleonora Rigoni
    • Sub-Investigator: Lara Ahmad
    • Sub-Investigator: Monica Bianchi
    • Sub-Investigator: Veronica Tosca
    • Sub-Investigator: Luciana Gracardi
    • Sub-Investigator: Romina Sarchi
    • Sub-Investigator: Anna Scarabotto
    • Sub-Investigator: Matteo Gastaldi
    • Sub-Investigator: Ghanya Al-Naqeb
    • Sub-Investigator: Federica Loperfido
    • Sub-Investigator: Mulubirhan A. Alemayohu
    • Sub-Investigator: Maria Cristina Monti
    • Sub-Investigator: Cristina Montomoli
    • Sub-Investigator: Danilo Porro
    • Sub-Investigator: Gloria Bertoli
    • Sub-Investigator: Alessia Lo Dico
    • Sub-Investigator: Clarissa Consolandi
    • Sub-Investigator: Marco Severgnini
    • Sub-Investigator: Francesca Costabile
    • Sub-Investigator: Stefano Decesari

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

• Sampling Method: Non-Probability Sample

Study Population

Persons with Multiple Sclerosis (pwMS) attending the Centro Sclerosi Multipla of the IRCCS Fondazione C. Mondino of Pavia will be eligible, according to the following inclusion criteria:

• Age ≥ 18 years
• Diagnosis of MS according to 2017 McDonald criteria
• Relapsing-remitting course (RR)
• Routine clinical examinations scheduled in the study period September 15, 2023, to March 15, 2024,
• Residing in the provinces of Pavia and Milano
• Informed consent form signed

Description

Inclusion Criteria:

• We will include persons with Multiple Sclerosis (pwMS) attending the Centro Sclerosi Multipla of the IRCCS Fondazione C. Mondino of Pavia, according to the following inclusion criteria:

  • Age ≥ 18 years
  • Diagnosis of MS according to 2017 McDonald criteria
  • Relapsing-remitting course (RR)
  • Routine clinical examinations scheduled in the study period September 15, 2023, to March 15, 2024,
  • Residing in the provinces of Pavia and Milano
  • Informed consent form signed

Exclusion Criteria:

• Exclusion criteria will involve people refusing to participate in the study or people not completely meet the inclusion criteria. Potentially eligible pwMS will be screened by a neurologist expert in MS who will verify that all the inclusion criteria will be fulfilled.

Study Plan

How is the study designed?

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort
Subjects with Multiple sclerosis; Age ≥ 18 years; Diagnosis of MS according to 2017 McDonald criteria; Relapsing-remitting (RR) MS; Residence in the provinces of Pavia or Milan (Italy); Informed consent form signed

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
microRNAs	The differential expression of a set of 7 specific circulating plasma microRNAs from the blood of each pwMS will be measured. The set of miRNAs has been already selected based on their significant deregulated expression in MS, as evidenced in a bioinformatic analysis conducted using computational resources. A portion of serum samples will be collected and immediately frozen at -20°C. After being transported, serum samples will be thawed and the circulating RNAs will be extracted. After a step of microRNA specific reverse transcription, the copy-DNA obtained will be used as the template for Real Time-qPCR analyses of selected miRNAs by Singleplex TaqMan microRNA assay reactions.The method of Livak et al. will be applied for the comparative analysis of the relative expression of the selected microRNAs in serum samples, considering different combinations of the external exposome variables.	from March 2024 to July 2024 visit
Air quality	Spatial information on pollutants such as PM10, PM2.5, PM1.0 expressed in microg/m3, nanoparticulate (n/cm3), and black carbon (microg/m3) from CAMS and CHIMBO databases and from Sentinel-5P TROPOMI will be included as gridded emissions in 0.1° x 0.1° (long-lat) resolution for the whole time series (daily values), from March 2023 onwards from the Pavia and Milano provinces. Large Eddy Simulations will be carried out over such areas with a high spatial-temporal resolution for estimating - in a quantitative manner - the 3D pollutants concentration at the street level.The grid will be further subdivided into the municipalities of the two provinces and a "personal exposure" will be also assessed, based on the geographic position of each pwMS derived from the residence address (geo-positioning).	from March 2023 to July 2023
Nutritional assessment	The dietary habits of the participants will be evaluated with the use of a validated food frequency questionnaire (FFQ) adapted to the Italian population and adjusted based on current food trends, such as the consumption of plant-based milk and meat. Detailed information about the daily food consumption of pwMS will be gained through a 24-h dietary recall.	from March 2024 to July 2024
Neurofilament (NfL)	Human NfL will be assessed on plasma samples by Simple Plex assay (ProteinSimple, CA, USA) on Ella device (ProteinSimple, CA, USA), according to the manufacturer's instructions. Calibration of Ella will be performed using the incartridge factory standard curve, and plasma samples will be measured with a 1:2 dilution in Sample Diluent (ProteinSimple, CA, USA). A single well was used for each sample as triplicates assays are automatically performed in Simple Plex assay microfluidic platform. The LLOQ is 2.70 pg/ml; the upper limit of quantification is 10,290.00 pg/ml.	from March 2024 to July 2024
Cytokines	Serum levels of IL1-beta, IL2, IL4, IL6, IL-10, IL12, IL17, IL22, IL23, TGF-beta, GM-CSF, TNF-alpha, IFN-gamma expressed in pg/mL will be assessed by using CXCL10 (performed using Simple Plex , R&D and commercial ELISA kit).	from March 2024 to July 2024
the Body Mass Index (BMI)	Weight in kg and height in m will be measured in order to calculate the Body Mass Index (BMI) as the ratio of weight (kg) to height (m) squared.	from March 2024 to July 2024
Physical activity	Physical activity levels will be evaluated by using the International Physical Activity Questionnaires (IPAQ) . The questionnaire is composed of 7 questions concerning the last 7 days levels of moderate, vigorous and sedentary physical activity. The metabolic equivalent of tasks (MET) will be calculated using the frequency and duration of physical activity and will be reported as MET-min/wk.	from March 2024 to July 2024
Quality of life related to MS	Quality of life will be evaluated by Multiple Sclerosis Quality of Life-29 (MSQOL-29), a multidimensional health-related quality of life measure that combines both generic and MS-specific items into a single instrument.	from March 2024 to July 2024
Sleep Quality	Sleep quality will be measured by the Pittsburgh Sleep Quality Index (PSQI) questionnaire. The questionnaire consists of 9 multiple-choice items divided into 5 different components related to subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbances, use of sleep medication and daytime dysfunction. The global PSQI score will be obtained by considering the scores of all the components and it ranges from 0 to 13. The PSQI cut-off of 5 or above indicates poor sleep quality.	from March 2024 to July 2024
Smoking habits	Cigarette smoking habits will be registered considering patients who are smoking, never smoked and quit smoking. The number of cigarettes smoked per day and the years of smoking will be recorded for smokers. Ex-smokers will be asked the number of years not smoking, the number of years that have smoked and the number of cigarettes smoked per day during their smoking years.	from March 2024 to July 2024
Dietary supplements use	Dietary supplements use will be assessed based on the type and nutrients included in the dietary supplements, their dose and their frequency of use.	from March 2024 to July 2024
Adherence to Mediterranean diet	Adherence to the Mediterranean dietary model will be assessed by applying the MEDI-LITE score. The MEDI-LITE score is based on a questionnaire, which investigates the frequency of consumption of nine classes of food. The final score varies from 0 (low adherence) to 18 (high adherence). Overall, subjects who will obtain a score higher than 9 will have a significantly increased adherence to the Mediterranean Diet.	from March 2024 to July 2024
Effect of diet on inflammation	The Dietary Inflammatory Index (DII) will be also calculated to quantify the overall effect of diet on inflammatory potential. The DII will be calculated based on the daily food consumption and then based on it the food parameters included in DII will be evaluated, such as vitamins, macronutrients, minerals, green/black tea and others.	from March 2024 to July 2024
waist and height ratio (WHtR)	The waist circumference and (cm) height (cm) will be measured in order to calculate their ratio (WHtR), a good predictor of abdominal adiposity in adults, will be also calculated.	from March 2024 to July 2024
Bioelectrical impedance	Bioelectrical impedance analysis will be used for the analysis of phase angle (PhA), which is considered an indicator of cellular health and malnutrition.	from March 2024 to July 2024

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Microbiome and biological biomarkers of oxidative stress and inflammation	The percentage of each bacterial operational taxonomic units (OTUs) was investigated separately for each sample, providing relative abundance data among the samples based on the the relative numbers of reads within each. Microbial profiles will be evaluated in terms of; Biodiversity (alpha-diversity), estimating the number of taxa in each sample and statistically comparing estimates across the experimental groups;; Composition (beta-diversity), evaluating the pairwise similarity among the samples and clustering them according to their distances;; Bacterial groups (relative abundances), evaluating whether a bacterial taxon is significantly increased or depleted in pwMS belonging to the different exposome variables under investigation.	from March 2024 to July 2024

Collaborators and Investigators

• Sponsor: University of Pavia
• Collaborators: IRCCS National Neurological Institute "C. Mondino" Foundation; National Research Council (Consiglio Nazionale delle Ricerche)
• Investigators: Principal Investigator: Eleonora Tavazzi, Neurological Institute-Foundation IRCCS Casimiro Mondino, Pavia, Italy

Study record dates

Study Major Dates

• Study Start (Estimated): April 15, 2024
• Primary Completion (Estimated): December 30, 2024
• Study Completion (Estimated): December 30, 2024

Study Registration Dates

• First Submitted: February 9, 2024
• First Submitted That Met QC Criteria: March 15, 2024
• First Posted (Actual): March 22, 2024

Study Record Updates

• Last Update Posted (Actual): March 22, 2024
• Last Update Submitted That Met QC Criteria: March 15, 2024
• Last Verified: October 1, 2023

More Information

Terms related to this study

• Keywords: physical activity; nutrition; microRNA; multiple sclerosis; inflammation; cytokines; microbiome; lifestyle; air pollution; exposome; neurofilament; urbanization
• Additional Relevant MeSH Terms: Pathologic Processes; Nervous System Diseases; Immune System Diseases; Demyelinating Autoimmune Diseases, CNS; Autoimmune Diseases of the Nervous System; Demyelinating Diseases; Autoimmune Diseases; Multiple Sclerosis; Sclerosis; Multiple Sclerosis, Relapsing-Remitting
• Other Study ID Numbers: 0040083/23

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

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