Multiple sclerosis (MS) is a chronic immune-mediated, multi factorial, neurodegenerative and inflammatory disease of the Central Nervous System (CNS).MS is a leading cause of neurological disability among young adults, especially women. The progressive cognitive impairment caused by the disease has a significant detrimental impact on the quality of life; for this reason it is very important to recognize it in time and to start adequate treatment at an early stage. Currently used disease-modifying therapies aim to reduce disease activity and progression and thus halt disability development. The evolution of the disease and the diagnosis of the progressive MS forms are monitored mainly using imaging techniques, that allow to highlight the presence of lesions in the gray matter and the loss of volume of the nervous system. To date, however, there are no universal criteria, especially in terms of biomarkers present in the blood, which is less invasive than the cerebrospinal fluid (CSF). The focal aim of this study was to contribute the field of biomarker discovery by identifying candidate protein biomarkers from plasma samples with the aim of gaining more insight into pathogenic aspects of MS. For this aim we used protein profile data obtained via antibody suspension bead affinity-based array technology from MS subjects and healthy related controls belonging to twenty multiplex families from the founder population of the Nuoro province, Sardinia (Italy).Plasma protein levels difference between MS cases and controls were estimated using statistical Linear Mixed Model(LMM) and by adding in the model the kinship matrix we also take into account for the familiar relationships between subjects. From results of the study we highlight the plasma protein complement 9 (C9) that could be regarded as putative diagnostic biomarker for MS as their level in plasma was significantly increased in MS cases compared to controls after multiple testing correction. Along this finding with the support of biological evidences, other up regulated or down regulated plasma proteins were identified: ZFP36L1, CD14, CYP24A1 and CP, SST, PLAT, CNTF. These proteins could be considered as “suggestive biomarkers”as they resulted non statistically significant after multiple testing corrections. In conclusion, our results provide indications for future plasma protein biomarker studies.
Analisi dei livelli di proteine plasmatiche nelle famiglie sarde di sclerosi multipla Alla ricerca di biomarcatori candidati. La Sclerosi Multipla (SM) è una malattia cronica immuno-mediata, multifattoriale, neurodegenerativa e infiammatoria del Sistema Nervoso Centrale (SNC). La SM è una delle principali cause di disabilità neurologica tra i giovani adulti, in articolare tra le donne. Ilprogressivo deterioramento cognitivo causato dalla malattia ha un notevole impatto negativo sullaqualità della vita; per questo motivo è molto importante riconoscerla in tempo e iniziare un trattamento adeguato in una fase precoce. Le terapie farmacologiche attualmente utilizzate mirano a ridurre la progressione della malattia e quindi arrestare lo sviluppo della disabilità. L’evoluzione della malattia e la diagnosi delle forme progressive di SM si basanosoprattutto su tecniche diimaging che consentono di evidenziare la presenza di lesioni a livello della sostanza grigia e la perdita di volume del tessuto nervoso.A tutt’oggi non esistono però dei criteri universali definitivi soprattutto in tema di biomarcatori presenti nel sangue, la cui raccolta è mena invasivo riispetto al liquido cerebrospinale (CSF).Lo scopo focale di questo studio è stato quello di contribuire all’identificazione di biomarcatori proteiciplasmaticicoinvolti nella patogenesi della SM. A questo scopo abbiamo analizzatoi livelli di alcune proteine plasmatichecandidate, misurate tramite l’antibodysuspension-bead array techonology,insoggetti affetti daSM e in controlli sani imparentati appartenenti in totale a venti famiglie multipledella provincia di Nuoro, Sardegna (Italia). Per stimare le differenze tra casi e controlli è stato utilizzato un modello lineare a effetti mistiche ha consentito, includendo nel modello la matrice di kinship, di tenere conto anche delle relazioni parentali esistenti tra i soggetti. Dopo correzione per test multipli, il risultato più rilevante è emerso perla proteina C9, il cui livello nel plasma è risultato significativamente aumentato nei casi rispetto ai controlli,che potrebbe avere quindi un ruolo come biomarcatore diagnostico per la SM. Altre proteine plasmatiche sono risultate up-o down-regolate nei casi rispetto ai controlli e sono: ZFP36L1, CD14, CYP24A1 e CP, SST, PLAT, CNTF. Queste proteine possono però solo essere indicate come “biomarcatori suggestivi”dato che perdono la significatività statistica dopo correzione per test multipli. Complessivamente i risultati ottenuti in questo lavoro possono fornire indicazioni per futuri studi sui biomarcatori plasmatici.
Plasma Protein Levels Analysis in Multiple Sclerosis Sardinian Families In search of Candidate Biomarkers
THIRUMOORTHI, VINEETHA
2021/2022
Abstract
Multiple sclerosis (MS) is a chronic immune-mediated, multi factorial, neurodegenerative and inflammatory disease of the Central Nervous System (CNS).MS is a leading cause of neurological disability among young adults, especially women. The progressive cognitive impairment caused by the disease has a significant detrimental impact on the quality of life; for this reason it is very important to recognize it in time and to start adequate treatment at an early stage. Currently used disease-modifying therapies aim to reduce disease activity and progression and thus halt disability development. The evolution of the disease and the diagnosis of the progressive MS forms are monitored mainly using imaging techniques, that allow to highlight the presence of lesions in the gray matter and the loss of volume of the nervous system. To date, however, there are no universal criteria, especially in terms of biomarkers present in the blood, which is less invasive than the cerebrospinal fluid (CSF). The focal aim of this study was to contribute the field of biomarker discovery by identifying candidate protein biomarkers from plasma samples with the aim of gaining more insight into pathogenic aspects of MS. For this aim we used protein profile data obtained via antibody suspension bead affinity-based array technology from MS subjects and healthy related controls belonging to twenty multiplex families from the founder population of the Nuoro province, Sardinia (Italy).Plasma protein levels difference between MS cases and controls were estimated using statistical Linear Mixed Model(LMM) and by adding in the model the kinship matrix we also take into account for the familiar relationships between subjects. From results of the study we highlight the plasma protein complement 9 (C9) that could be regarded as putative diagnostic biomarker for MS as their level in plasma was significantly increased in MS cases compared to controls after multiple testing correction. Along this finding with the support of biological evidences, other up regulated or down regulated plasma proteins were identified: ZFP36L1, CD14, CYP24A1 and CP, SST, PLAT, CNTF. These proteins could be considered as “suggestive biomarkers”as they resulted non statistically significant after multiple testing corrections. In conclusion, our results provide indications for future plasma protein biomarker studies.È consentito all'utente scaricare e condividere i documenti disponibili a testo pieno in UNITESI UNIPV nel rispetto della licenza Creative Commons del tipo CC BY NC ND.
Per maggiori informazioni e per verifiche sull'eventuale disponibilità del file scrivere a: unitesi@unipv.it.
https://hdl.handle.net/20.500.14239/15104