Background: Necrotizing enterocolitis (NEC) is the most devastating gastrointestinal emergency of the preterm neonate. Given its multifaceted pathophysiology, the search for early predictive biomarkers of NEC remains challenging. Metabolomics, the last of the four big technology-based omic sciences, may allow the identification of known and unknown metabolites involved in the molecular processes responsible for NEC. Materials and Methods: This was a monocentric observational case-control study applying the untargeted metabolomic approach on urine samples of preterm infants born <34 gestational weeks (GW) to identify early predictive profiles of NEC development and to discriminate NEC cases from healthy controls and neonates developing spontaneous intestinal perforation (SIP). Neonates had their urine collected at birth (within 48 hours, T0), at 14 days (T1) and at 28 days of life (T2). Untargeted metabolomic analysis was performed with Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). The framework of Model Population Analysis (MPA), as well as univariate and multivariate statistical analyses, were used to analyze data and compare the different populations. A p value of 0.05 was considered statistically significant. Results: Twenty NEC cases, 17 healthy controls, 3 subjects with SIP and 7 subjects with other gastrointestinal diseases were similar for the main clinical prenatal and postnatal characteristics. Considering urine samples collected at birth, MPA was able to correctly predict 16 out of 20 subjects developing NEC (80%), 5 out of 7 subjects with other gastrointestinal pathologies (71.4%), and all the subject developing SIP (100%) as not belonging to the control group. This highlights that the urinary metabolome is closely associated to the disease under investigation. Metabolomic analysis was able to distinguish the NEC Group from the Control Group both at T0 and by the evolution of urine samples over time, proving that part of the differences observed at T0 are maintained until 28 days. The exploratory analysis comparing medical vs surgical NEC cases (7 vs 7 subjects) showed a clear separation between the two groups at T0, with lower 5-hydroxyindolacetic acid and higher N-acetylaspartic acid, butyrylcarnitine and propionylcarnitine in surgical cases. Conclusions: Our study shows that metabolomics applied on very early urine samples (first 2 days of life) is able to discriminate neonates developing NEC and those developing SIP or other gastrointestinal conditions from healthy controls. Metabolic differences between NEC cases and controls at birth persist during the first month of life. Additionally, NEC cases requiring surgery are metabolically different from those treated medically from the very early urine samples. In our study, potential predictive biomarkers of NEC development and severity mainly play a role in pathways of oxidative stress. These results need to be validated in future studies on independent cohorts using targeted analysis.
Introduzione: L'enterocolite necrotizzante (NEC) è l'emergenza gastrointestinale più devastante del neonato pretermine. Data la sua complessa e sfaccettata fisiopatologia, la ricerca di biomarker predittivi precoci di NEC resta una sfida. La metabolomica, l'ultima delle quattro grandi scienze omiche basate sulla tecnologia, potrebbe permettere l'identificazione di metaboliti conosciuti e ancora sconosciuti coinvolti nei processi molecolari responsabili della NEC. Materiali e metodi: Questo è uno studio caso-controllo osservazionale monocentrico che ha applicato l'approccio metabolomico untarget su campioni di urina di neonati pretermine nati con età gestazionale alla nascita <34 settimane gestazionali per identificare profili predittivi precoci di sviluppo di NEC e per discriminare i casi di NEC da controlli sani e da neonati che sviluppano perforazione intestinale spontanea (SIP). Sono stati raccolti campioni urinari di neonati alla nascita (entro 48 ore, T0), a 14 giorni (T1) e a 28 giorni di vita (T2). L'analisi metabolomica untarget ha utilizzato l'Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). La struttura della Model Population Analysis (MPA), così come l'analisi statistica univariata e multivariata, sono state applicate per l'analisi dei dati e per confrontare le diverse popolazioni. Un valore p di 0.05 è stato considerato statisticamente significativo. Risultati: Venti casi di NEC, 17 controlli sani, 3 soggetti con SIP e 7 soggetti con altre patologie gastrointestinali sono risultati simili per le principali caratteristiche cliniche prenatali e postnatali. Considerati i campioni urinari raccolti alla nascita, la MPA è stata in grado di predire correttamente 16 su 20 (80%) soggetti con sviluppo di NEC, 5 su 7 (71.4%) soggetti con altre patologie gastrointestinali, e tutti i soggetti con sviluppo di SIP (100%) come non appartenenti al gruppo di controlli sani. Questo sottolinea come il metaboloma urinario sia strettamente associato alla patologia sottostante. L'analisi metabolomica ha permesso di distinguere il gruppo NEC dal gruppo Controlli sia a T0 che dall'evoluzione dei campioni urinari nel tempo, dimostrando che parte delle differenze osservate a T0 si mantengono a 28 giorni. L'analisi esplorativa che ha confrontato casi di NEC medica da casi di NEC chirurgica (7 vs 7 soggetti) ha mostrato una chiara separazione dei due gruppi a T0, con ridotti livelli di acido 5-idrossiindolacetico e aumentati livelli di acido N-acetilaspartico, butirilcarnitina e propionilcarnitina nei casi di NEC chirurgica. Conclusioni: Il nostro studio mostra the la metabolomica applicata a campioni urinari molto precoci (entro due giorni di vita) è capace di discriminare i neonati che sviluppano NEC, quelli che sviluppano SIP o altre condizioni gastrointestinali dai controlli sani. Le differenze metaboliche tra casi di NEC e controlli alla nascita persistono durante il primo mese di vita. Inoltre, i casi di NEC con necessità di intervento chirurgico sono metabolicamente diversi da quelli trattati con terapia medica già dai campioni urinari precoci. Nel nostro studio, i potenziali biomarcatori predittivi di sviluppo e di severità di NEC giocano un ruolo nelle vie di stress ossidativo. Questi risultati devono essere validati in studi futuri su coorti indipendenti utilizzando l'analisi target.
ANALISI METABOLOMICA NELLA PREDIZIONE DELL'INSORGENZA E DELLA SEVERITA' DELL'ENTEROCOLITE NECROTIZZANTE / Moschino, Laura. - (2024 Jan 18).
ANALISI METABOLOMICA NELLA PREDIZIONE DELL'INSORGENZA E DELLA SEVERITA' DELL'ENTEROCOLITE NECROTIZZANTE
MOSCHINO, LAURA
2024
Abstract
Background: Necrotizing enterocolitis (NEC) is the most devastating gastrointestinal emergency of the preterm neonate. Given its multifaceted pathophysiology, the search for early predictive biomarkers of NEC remains challenging. Metabolomics, the last of the four big technology-based omic sciences, may allow the identification of known and unknown metabolites involved in the molecular processes responsible for NEC. Materials and Methods: This was a monocentric observational case-control study applying the untargeted metabolomic approach on urine samples of preterm infants born <34 gestational weeks (GW) to identify early predictive profiles of NEC development and to discriminate NEC cases from healthy controls and neonates developing spontaneous intestinal perforation (SIP). Neonates had their urine collected at birth (within 48 hours, T0), at 14 days (T1) and at 28 days of life (T2). Untargeted metabolomic analysis was performed with Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). The framework of Model Population Analysis (MPA), as well as univariate and multivariate statistical analyses, were used to analyze data and compare the different populations. A p value of 0.05 was considered statistically significant. Results: Twenty NEC cases, 17 healthy controls, 3 subjects with SIP and 7 subjects with other gastrointestinal diseases were similar for the main clinical prenatal and postnatal characteristics. Considering urine samples collected at birth, MPA was able to correctly predict 16 out of 20 subjects developing NEC (80%), 5 out of 7 subjects with other gastrointestinal pathologies (71.4%), and all the subject developing SIP (100%) as not belonging to the control group. This highlights that the urinary metabolome is closely associated to the disease under investigation. Metabolomic analysis was able to distinguish the NEC Group from the Control Group both at T0 and by the evolution of urine samples over time, proving that part of the differences observed at T0 are maintained until 28 days. The exploratory analysis comparing medical vs surgical NEC cases (7 vs 7 subjects) showed a clear separation between the two groups at T0, with lower 5-hydroxyindolacetic acid and higher N-acetylaspartic acid, butyrylcarnitine and propionylcarnitine in surgical cases. Conclusions: Our study shows that metabolomics applied on very early urine samples (first 2 days of life) is able to discriminate neonates developing NEC and those developing SIP or other gastrointestinal conditions from healthy controls. Metabolic differences between NEC cases and controls at birth persist during the first month of life. Additionally, NEC cases requiring surgery are metabolically different from those treated medically from the very early urine samples. In our study, potential predictive biomarkers of NEC development and severity mainly play a role in pathways of oxidative stress. These results need to be validated in future studies on independent cohorts using targeted analysis.File | Dimensione | Formato | |
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