{"id":37954,"date":"2025-03-07T09:35:12","date_gmt":"2025-03-07T07:35:12","guid":{"rendered":"https:\/\/umfst.ro\/?post_type=proiect-european&#038;p=37954"},"modified":"2025-03-17T11:50:37","modified_gmt":"2025-03-17T09:50:37","slug":"rolul-microbiotei-in-evolutia-clinica-a-pacientilor-cu-scleroza-multipla-tratati-cu-interferon-beta-o-abordare-complexa","status":"publish","type":"proiect-european","link":"https:\/\/umfst.ro\/fr\/universitate\/departament-proiecte-europene\/proiect-european\/rolul-microbiotei-in-evolutia-clinica-a-pacientilor-cu-scleroza-multipla-tratati-cu-interferon-beta-o-abordare-complexa\/","title":{"rendered":"Rolul microbiotei \u00een evolu\u021bia clinic\u0103 a pacien\u021bilor cu Scleroz\u0103 Multipl\u0103 trata\u021bi cu interferon-beta: o abordare complex\u0103"},"content":{"rendered":"<div class=\"wp-block-columns is-layout-flex wp-container-core-columns-is-layout-28f84493 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\" style=\"flex-basis:100%\">\n<figure class=\"wp-block-table\"><table class=\"has-fixed-layout\"><tbody><tr><td>Proposal Registration Code<\/td><td>PN-III-P1-1.1-PD-2021-0547<\/td><\/tr><tr><td>Project Registration Code<\/td><td>PD 80\/2022<\/td><\/tr><tr><td>Project Acronym<\/td><td>Micro-MS<\/td><\/tr><tr><td>Purpose and project planning<\/td><td>By considering the gut-brain axis from four perspectives &#8211; immunity (Th1, Th2, Th17), local microbiota products (short-chain fatty acids -SCFA), vagus nerve signalling and neuroprotection (brain derived neurotrophic factor &#8211; BDNF) and loss of blood-brain barrier (BBB) integrity by matrix metalloproeinases (MMP), complex biological implications can be established between the microbiome and neuroinflammation. This complex quadri-dimensional approach based on a clinical longitudinal MS population has not been considered in clinical terms before. The direct intestinal microbiome is ephemeral and highly influenced by alimentation, therefore, plasma SCFA analysis will eliminate the most frequent bias. The project will use a highly selective QTOF LC-MS system with a purpose-developed method in order to measure plasma SCFA levels. LC-MS is a modern technique increasingly used in recent years for biomonitoring and diagnostics.Project plan:Socio-demographical and clincal assessment of study population (20 RRMS, 20 SPMS, 20 benign MS, 20 naive MS, 20 controls);Harvesting of the plasma samples;SCFA analysis by HPLC (plasma): Development of detection of acetic, propionic, butyric and caproic acids and internal standard without and after derivatization, development of HPLC separation of the analytes, development of extraction method of the analytes from plasma, performance verification of the proposed analytical method: linearity, accuracy and precision;MMP and cytokine analysis by xMAP technology. Color-coded internally-dyed magnetic beads coated with specific monoclonal antibodies bind the analytes of interest in the biological sample. After unbound antigens are washed away, the previously formed antigen-antibody complexes are tagged with a fluorescent reporter via streptavidin\/avidin-biotin bridges. Analyte concentrations are determined by a 2-laser interrogation system: red laser for specificity identification based on each bead\u2019s internal dye, and green laser for quantification of bound analytes based on reporter fluorescence intensities. Patient samples with unknown concentrations are evaluated based on a calibration curve generated in the same analytical run;BDNF by ELISA method.<\/td><\/tr><tr><td>Project Start Date<\/td><td>01.04.2022<\/td><\/tr><tr><td>Project End Date<\/td><td>01.04.2024<\/td><\/tr><tr><td>Project Duration<\/td><td>24 months<\/td><\/tr><tr><td>Total Budget Value<\/td><td>50.000 EUR<\/td><\/tr><tr><td>Funding<\/td><td>Ministry of Research and Innovation, CNCS-UEFISCDI<\/td><\/tr><tr><td>Main Objectives<\/td><td><strong>Main Objective:<\/strong> To determine the influence that plasma SCFA have upon the peripheral immune processes (a selected cytokine pannel), BBB permeability (MMP), vagus nerve signalling and neuroprotection (BDNF) in different groups of MS patients (RRMS, SPMS, BMS, na\u00efve MS phenotype) treated with IFN\u03b2; Specific <strong>Objectives:<\/strong>(1) Development of detection of acetic, propionic, BA, CA and improve the internal standard of&nbsp; chromatography and mass spectrometry laboratory within the Centre for Advanced Medical and Pharmaceutical Research (CCAMF), without and after derivatization by high performance liquid chromatography (HPLC) in MS patients and HC;(2) To assess the impact of microbiota in clinical evolution of MS patients treated with IFN\u03b2;(3) To evaluate the dietary characteristics of the target population by personalised questionnaire;<\/td><\/tr><tr><td>Project Director<\/td><td>Laura-Iulia Barcutean<\/td><\/tr><tr><td>Project Mentor<\/td><td>Rodica Balasa<\/td><\/tr><tr><td>Results<\/td><td>The development of an effective method to evaluate plasma SCFA in MS patients using HPLC in order to discover if&nbsp; SCFA levels as biomarkers targeted for MS population can be used as biomarkers.<\/td><\/tr><tr><td>Host Institution<\/td><td>\u201eGeorge Emil Palade\u201d University of Medicine, Pharmacy, Science and Technology of Targu Mures<\/td><\/tr><\/tbody><\/table><\/figure>\n<\/div>\n<\/div>\n\n\n\n<p><br><strong>Results:<\/strong><\/p>\n\n\n\n<p>&nbsp;<strong>Achieved within 1\/2022 stage<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Coordination of a <strong>bachelor&#8217;s thesis<\/strong> of the student Manea Andrei, Faculty of General Medicine, entitled <strong>&#8222;Implications of nutrition in the evolution of patients with multiple sclerosis&#8221;,<\/strong> a thesis that was supported in the July 2022 Session, final grade 10 (Appendix 1).<\/li>\n\n\n\n<li>The organization of a special session entitled:<strong> &#8222;Microbiome alterations and implications in patients with multiple sclerosis \u2013 targeting neuroinflammation from the intestinal compartment&#8221;<\/strong>, within the first edition of the International Conference Toward Precision Medicine in Multiple Sclerosis Management, which took place in T\u00e2rgu Mure\u0219 during the 7 &#8211; October 8, 2022; with works presented in abstract indexed BDI, ISSN 2971-8171 (Appendix 2).<\/li>\n<\/ul>\n\n\n\n<p><strong>Laura B\u0103rcu\u021bean<\/strong> (Project manager): <em>Potential mechanisms of immune modulation targeting the intestinal microbiota.<\/em><\/p>\n\n\n\n<p><strong>Rodica B\u0103la\u0219a<\/strong> (mentor): <em>Gut-Brain axis: is MS a digestive triggered disease?<\/em><\/p>\n\n\n\n<p><strong>Achieved within 2\/2023 stage<\/strong><\/p>\n\n\n\n<p>Supporting a workshop with the theme: <strong>&#8222;Gut-Brain Axis. The hidden agenda of autoimmune disorders \u2013 focus on multiple sclerosis&#8221;<\/strong> during the 27th edition of the International Congress for Students, Young Doctors and Pharmacists, Marisiensis, May 17-21, 2023, T\u00e2rgu Mure\u0219 (Appendix 1)<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Active participation in the 10th <strong>International Conference &#8222;Targeting Microbiota \u2013 Towards Clinical Revolution&#8221; <\/strong>which took place between October 17-19, 2023 in Venice, Italy; Linkedin (Abstract Annex 2)<\/li>\n\n\n\n<li>Abstract accepted, presentation in progress at the <strong>Scientific Session of University Staff within the UMFST Days<\/strong>, an event that will take place on December 14-15, 2023 in T\u00e2rgu Mure\u0219, Romania; Link (Appendix 3)<\/li>\n\n\n\n<li>Completed review article (Title: <strong>Investigating the immunomodulatory potential of short-chain fatty acids in multiple sclerosis<\/strong>), submitted for evaluation to a profile journal Q1 (Summary graphic representation of the properties of butyrate, original image from the article in Appendix 4)<\/li>\n<\/ul>\n\n\n\n<p><strong>Achieved within the project (01.04.2022 \u2013 31.03.2024)<\/strong><\/p>\n\n\n\n<p><strong>Summary of the project\u2019s theme<\/strong><\/p>\n\n\n\n<p>According to the hygiene hypothesis, the <strong>increasing prevalence and incidence of autoimmune diseases such as multiple sclerosis (MS)<\/strong>, particularly in industrialized countries, results from an underdeveloped immune system during the formative years. Although the studies in this aspect have not established a clear connection for the pathogenesis of MS, it prompted <strong>the investigations in intestinal microbiota and the fermentation metabolites of the commensal bacteria<\/strong>. The gut bacteria ferment dietary fibers to produce <strong>short and medium fatty acids (SCFA\/MCFA).<\/strong> <strong>SCFA<\/strong> have demonstrated <strong>immunomodulatory properties<\/strong> and can transverse the blood-brain barrier and hypothetically exert their effects inside the central nervous system, by modulating inflammation and promoting neuroplasticity and remyelination. In the periphery, SCFA promote the differentiation of <strong>regulatory T lymphocytes<\/strong> and secretion of <strong>anti-inflammatory cytokines such as interleukin \u2013 10<\/strong>. <strong>MCFA, such as caproic acid<\/strong>, have contrasting, pro-inflammatory effects depending on the microbial pattern of the patients. Caproic acid promotes the immune differentiation toward a <strong>pro-inflammatory phenotype, Th1 and Th17<\/strong>, with secretion of proinflammatory cytokines. The selective permeability of the blood brain barrier ensures that immune cells have no way of passage into the brain parenchyma. In MS, the <strong>blood-brain barrier permeability is altered <\/strong>and inflammatory cells pass through, exerting the inflammatory cascade leading to demyelination, neurodegeneration and axonal loss. Matrix metalloproteinases (MMPs) are a set of enzymes involved in the maintenance and structure of the endothelial cells that compose the blood-brain barrier. MMP9 in particular has been linked with <strong>destructive effects<\/strong> on the blood-brain barrier and also the intestinal barrier.<\/p>\n\n\n\n<p>The gut brain axis represents a d<strong>ynamic communication between the intestinal tract and the central nervous system<\/strong>, functioning as a bidirectional signaling pathway through which the enteric system influences the brain and vice-versa. The central element of this ecosystem is represented by the intestinal microbiota. The direct connection between the intestinal compartment and the brain is partially <strong>mediated via the vagus nerve<\/strong>. One of the most important neuromodulators involved in this bi-directional communication is <strong>brain-derived neurotrophic factor (BDNF),<\/strong> which modulates the excitability of the vagal neurons. BDNF is also highly influenced by the SCFA, particularly butyric acid. The experimental approach of the intestinal microbiota is challenging due to the multitude of short-term influencing factors such as alcohol consumption, diet, treatment with pre\/pro biotics, stress, unhealthy diet, etc.<\/p>\n\n\n\n<p>Therefore, the present project proposed <strong>two important directions of research<\/strong>:<\/p>\n\n\n\n<ol class=\"wp-block-list\">\n<li><strong>The analysis of inflammation biomarkers from a quadri dimensional perspective (see schematic below)<\/strong>:\n<ul class=\"wp-block-list\">\n<li>Intestinal compartment: assessment of peripheral levels of SCFA\/MCFA<\/li>\n\n\n\n<li>Interaction between the gut and the brain via the vagus nerve: BDNF<\/li>\n\n\n\n<li>Pro and anti-inflammatory cytokines<\/li>\n\n\n\n<li>Integrity of the blood-brain barrier: MMP<\/li>\n<\/ul>\n<\/li>\n<\/ol>\n\n\n\n<figure class=\"wp-block-image\"><img decoding=\"async\" width=\"217\" height=\"208\" src=\"https:\/\/umfst.ro\/wp-content\/uploads\/2024\/03\/2proiect.png\" alt=\"\" class=\"wp-image-23918\" srcset=\"https:\/\/umfst.ro\/wp-content\/uploads\/2024\/03\/2proiect.png 217w, https:\/\/umfst.ro\/wp-content\/uploads\/2024\/03\/2proiect-13x12.png 13w\" sizes=\"(max-width: 217px) 100vw, 217px\" \/><\/figure>\n\n\n\n<ol start=\"2\" class=\"wp-block-list\">\n<li><strong>Implementing and validating a working method by liquid chromatography-mass spectrometry<\/strong> (LC-MS\/MS) <strong>to assess SCFA and MCFA<\/strong> from serum samples<\/li>\n<\/ol>\n\n\n\n<p><strong>Concrete achievements<\/strong><\/p>\n\n\n\n<ol style=\"list-style-type:upper-roman\" class=\"wp-block-list\">\n<li><strong>Implementation and validation<\/strong> of a working method for LC-MS\/MS which allowed for the analysis of <strong>SCFA <\/strong>and<strong> MCFA<\/strong> from human serum, namely <strong>acetic acid, propionic acid, butyric acid and caproic acid<\/strong>, metabolites of the human microbiome with demonstrated immunomodulatory effects in MS. The method was developed and applied in the Center for Advanced Medical and Pharmaceutical Research, CCAMF (<a href=\"https:\/\/eeris.eu\/ERIF-2000-000C-0801\" target=\"_blank\" rel=\"noopener\">https:\/\/eeris.eu\/ERIF-2000-000C-0801<\/a>) from the George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures.<\/li>\n<\/ol>\n\n\n\n<p>The development and validation of a <strong>selective, sensitive, precise, and accurate LC-MS method <\/strong>within this project enable the conduct of various types of <strong>future clinical, preclinical, biomonitoring, or metabolomic<\/strong> studies at reduced costs and with short analysis times, also allowing for efficient planning of such studies.<\/p>\n\n\n\n<ol start=\"2\" style=\"list-style-type:upper-roman\" class=\"wp-block-list\">\n<li><strong>Publications:<\/strong> <strong>(1)<\/strong> ISI-Q1 (published &#8211; review): <strong>Barcutean L,<\/strong> Maier S, Burai-Patrascu M, Farczadi L, <strong>Balasa R.<\/strong> The Immunomodulatory Potential of Short-Chain Fatty Acids in Multiple Sclerosis. Int J Mol Sci. 2024; 25(6):3198. https:\/\/doi.org\/10.3390\/ijms25063198. FI: 5.6.; <strong>(2)<\/strong> ISI-Q2 (published \u2013 original research): <strong>Barcutean L,<\/strong> Maier S, Bajko Z, Stoian A, Mosora O, Sarmasan Emanuela, Manescu I-B, <strong>Balasa R.<\/strong> Long-Term Disability Outcomes in Relapsing\u2013Remitting Multiple Sclerosis Patients: Impact of Clinical and Demographic Factors on Disease Progression. J Clin Med. 2024; 13:1813. https:\/\/doi.org\/10.3390\/jcm13061813, IF: 3.9.; <strong>(3)<\/strong> ISI-Q1 (under review): sent to IJMS, manuscript ID 2956963, title: Peripheral fatty acids dynamics in response to interferon beta treatment in na\u00efve multiple sclerosis patients: insights into inflammation, gut-brain connection and blood-brain permeability.<\/li>\n\n\n\n<li><strong>Organisation of a special session dedicated to microbiota within an international conference:<\/strong><\/li>\n\n\n\n<li>\n<ul class=\"wp-block-list\">\n<li><strong>Microbiome alterations and implications in patients with multiple sclerosis \u2013 targeting neuroinflammation from the intestinal compartment<\/strong>; International Conference: <em>Toward Precision Medicine in Multiple Sclerosis Management<\/em>, 7-8 Oct 2022, Targu Mures, Romania\n<ul class=\"wp-block-list\">\n<li>Presented papers:\n<ol class=\"wp-block-list\">\n<li><strong>Rodica Balasa:<\/strong> Gut-brain axis: is MS a digestive triggered disease?<\/li>\n\n\n\n<li><strong>Anca Negovan:<\/strong> Environmental risk factors altering the intestinal microbiota<\/li>\n\n\n\n<li><strong>Monica Tarcea, Gliga Florina:<\/strong> Diet and gut microbiota implications in multiple sclerosis<\/li>\n\n\n\n<li><strong>Laura Barcutean, Lenard Farczadi, Rodica Balasa:<\/strong> Potential mechanisms of immune modulation targeting the intestinal microbiota<\/li>\n\n\n\n<li><strong>Adina Hutanu:<\/strong> Inflammatory biomarkers for microbiota dysregulation in multiple sclerosis<\/li>\n\n\n\n<li><strong>Farczadi Lenard, Laura Barcutean, Silvia Imre:<\/strong> The role of liquid chromatography-mass spectrometry in modern medicine \u2013 assessing the impact of microbiota by short-chain fatty acids assay<\/li>\n<\/ol>\n<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<\/li>\n\n\n\n<li><strong>Active<\/strong> presentations in national and international conferences in the field of research:\n<ul class=\"wp-block-list\">\n<li>Oral presentation in the 10<sup>th<\/sup> World Congress of the International Society of Microbiota: Targeting Microbiota, Towards Clinical Revolution, Venice, 17-19 Oct, Italy with the paper: \u201c<strong>The role of microbiome in multiple sclerosis: a multidimensional approach<\/strong>\u201d<\/li>\n\n\n\n<li>Oral presentation in the Scientific Session of University Academic Staff with the paper \u201c<strong>The role of brain-derived neurotrophic factor in interferon beta-treated multiple sclerosis patients<\/strong>\u201d, 11-15 Dec, Targu Mures, Romania<\/li>\n\n\n\n<li>Accepted abstract (poster, will be presented) in the 10<sup>th<\/sup> Congress of the European Academy of Neurology, 29 Jun \u2013 2 July, Helsinki, Finland with the paper: \u201c<strong>Brain-derived neurotrophic factor and progression in multiple sclerosis.<\/strong>\u201d<\/li>\n\n\n\n<li>Accepted abstract (oral presentation) in the The 22nd Congress of the Romanian Society of Neurology, 5-7 June 2024 with the paper: \u201c<strong>Microbiota and Multiple Sclerosis<\/strong>\u201d.<\/li>\n<\/ul>\n<\/li>\n<\/ol>\n\n\n\n<ol start=\"5\" style=\"list-style-type:upper-roman\" class=\"wp-block-list\">\n<li>Student\u2019s master and bachelor\u2019s degree\n<ul class=\"wp-block-list\">\n<li>I coordinated the bachelor\u2019s degree for the student <strong>Andrei Manea<\/strong>, Faculty of General Medicine, within George Emil Palade University of Medicine, Pharmacy, Science and Technology, with the title <em><strong>\u201cImplications of nutrition in the evolution of patients with multiple sclerosis\u201d.<\/strong><\/em>The thesis was presented in July 2022, final grade 10.<\/li>\n\n\n\n<li>I am coordinating a master\u2019s thesis for the student <strong>Andreea Kelemen,<\/strong> master student in the program Clinical and Community Nutrition, within the George Emil Palade University of Medicine, Pharmacy, Science and Technology, with the title \u201c<em><strong>Dietary Considerations for Individuals with Multiple Sclerosis&#8221;.<\/strong><\/em><\/li>\n<\/ul>\n<\/li>\n\n\n\n<li>Student dedicated workshop\n<ul class=\"wp-block-list\">\n<li>I coordinated the first workshop for medical students within the XXVII<sup>th<\/sup> International Congress for Students, Young Doctors and Pharmacists, Mariensis, in Targu Mures regarding the implications of gut-brain axis in demyelinating diseases, with a focus on MS, entitled: <strong>\u201cGut-brain axis. The hidden agenda of autoimmune disorders \u2013 focus on multiple sclerosis\u201d<\/strong>, which took place on 17-21 may 2023, Targu Mures, Romania<\/li>\n\n\n\n<li>The 2<sup>nd<\/sup> workshop entitled \u201c<strong>Microbiota in multiple sclerosis: truth or dare?\u201d<\/strong> has been announced and will be held on 23<sup>th<\/sup> of May, in Targu Mures, Romania, within the XXIII<sup>th <\/sup>International Congress for Students, Young Doctors and Pharmacists, Marisiensis.<\/li>\n<\/ul>\n<\/li>\n<\/ol>","protected":false},"excerpt":{"rendered":"<p>Proposal Registration Code PN-III-P1-1.1-PD-2021-0547 Project Registration Code PD 80\/2022 Project Acronym Micro-MS Purpose and project planning By considering the gut-brain axis from four perspectives &#8211; immunity (Th1, Th2, Th17), local microbiota products (short-chain fatty acids -SCFA), vagus nerve signalling and neuroprotection (brain derived neurotrophic factor &#8211; BDNF) and loss of blood-brain barrier (BBB) integrity by [&hellip;]<\/p>\n","protected":false},"featured_media":38345,"template":"","meta":{"_relevanssi_hide_post":"","_relevanssi_hide_content":"","_relevanssi_pin_for_all":"","_relevanssi_pin_keywords":"","_relevanssi_unpin_keywords":"","_relevanssi_related_keywords":"","_relevanssi_related_include_ids":"","_relevanssi_related_exclude_ids":"","_relevanssi_related_no_append":"","_relevanssi_related_not_related":"","_relevanssi_related_posts":"20957,37749,37266,36881,24107,37471","_relevanssi_noindex_reason":"","footnotes":""},"sursa-finantare":[251],"class_list":["post-37954","proiect-european","type-proiect-european","status-publish","has-post-thumbnail","hentry","sursa-finantare-fonduri-nationale"],"meta_box":{"domeniu":"s\u0103n\u0103tate","valoare_proiect":"50.000 EUR","director_de_proiect":"Laura-Iulia Barcutean","perioada_de_implementare":"01.04.2022 - 01.04.2024","website":""},"_links":{"self":[{"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/proiect-european\/37954","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/proiect-european"}],"about":[{"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/types\/proiect-european"}],"version-history":[{"count":0,"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/proiect-european\/37954\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/media\/38345"}],"wp:attachment":[{"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/media?parent=37954"}],"wp:term":[{"taxonomy":"sursa-finantare","embeddable":true,"href":"https:\/\/umfst.ro\/fr\/wp-json\/wp\/v2\/sursa-finantare?post=37954"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}