Ocenjevanje mesta odvzema biopsij pri aktivni Crohnovi bolezni

NOVAK, GREGOR (author), Štabuc, Borut (mentor)

Izhodišča in namen: Histološka remisija je možen terapevtski cilj pri zdravljenju bolnikov s Crohnovo boleznijo (CB), vendar do danes ni določenih meril za odvzem biopsij črevesne sluznice. Namen naše raziskave je določiti mesto za odvzem biopsij v prisotnosti razjed pri CB, ki daje najvišje histopatološko točkovanje, največjo sluznično mRNK izražanje pro-vnetnih genov in največje število vnetnih celic. Hipoteza in cilji: Naša hipoteza je, da obstajajo razlike v histopatoloških točkovanjih, izražanju genov in številu vnetnih celic v biopsijah črevesne sluznice na različnih razdaljah od razjed pri CB. Cilji študije so določitev mesta odvzema biopsij pri CB, pridobljenih s treh različnih razdalj od razjed v debelem in tankem črevesu, ki ima: • najvišjo histopatološko točkovanje z uporabo več točkovnikov histološke aktivnosti bolezni, • najvišjo izražanje mRNK izbranih genov, • največje število vnetnih celic, dokazanih imunohistokemično. Metode: V prospektivno multicentrično študijo v treh centrih smo vključili 51 bolnikov s histološko potrjeno diagnozo CB in z razjedami > 0,5 cm v debelem in/ali tankem črevesu. Biopsije smo odvzeli na treh različnih razdaljah od razjede. Histopatološko aktivnost smo ocenili s tremi točkovniki histološke aktivnosti bolezni. Izražanje mRNK petih pro-vnetnih genov smo določili s kvantitativno verižno reakcijo s polimerazo. Število vnetnih celic v lamini proprii smo določili z imunohistokemičnim barvanjem. Za statistično analizo smo uporabili modele mešanih učinkov. Rezultati: Potrdili smo hipotezo, da so prisotne razlike v histopatoloških točkovanjih, izražanju pro-vnetnih genov in številu MPO-pozitivnih celic v biopsijah črevesne sluznice na različnih razdaljah od razjede pri CB. Biopsije odvzete iz roba razjede imajo večjo histopatološko vnetno aktivnost, izražanje mRNK pro-vnetnih genov in število MPO-pozitivnih celic v lamini proprii v primerjavi z biopsijami odvzetimi v bližini ulkusa tako v debelem kot tankem črevesju (p<0,005). Zaključki: Mesto odvzema biopsije pri CB, ki doseže največjo histopatološko vnetno aktivnost bolezni in najvišje sluznično izražanje mRNK pro-vnetnih genov, je rob ulkusa.

• Language: Slovenian
• Keywords: Crohnova bolezen, biopsija, histologija, histološka remisija, transkriptomika
• Work type: Doctoral dissertation
• Organization: MF - Faculty of Medicine
• Year: 2019
• PID: 20.500.12556/RUL-111990
• COBISS.SI-ID: 302925568
• Publication date in RUL: 19.10.2019

Secondary language

• Language: English
• Title: Evaluating location of biopsy procurement in active Crohn's diease

Abstract

Background and aim: Histological remission is a potential therapeutic target in Crohn’s disease (CD) but no standards exist to guide the sampling of mucosal biopsies. We aim to determine the location for biopsy procurement in the presence of ulcers which yields the highest histopathological score, the greatest mRNA expression of pro-inflammatory genes and the highest number of inflammatory cells. Hypothesis and objectives: Our hypothesis is that differences in histopathological scores, mucosal gene expression and number of inflammatory cells of mucosal biopsies at various distances from ulcers in CD exist. The objective of the study is to determine the location of biopsy, procured from three different distances from ulcers in the colon and ileum, which yields: • the highest histopathological scores using several indices • the greatest mucosal mRNA expression of selected genes • the highest number of inflammatory cells as detected by immunohistochemistry. Methods: This prospective multicentric study enrolled 51 patients from three centers with a histologically confirmed diagnosis of CD and ulcers > 0.5 cm in the colon or/and ileum. Biopsy specimens were obtained at three different distances from ulcers. Histopathological disease activity was assessed using three histological scores. mRNA expression was measured by quantitative polymerase chain reaction. Number of inflammatory cells was determined by immunohistochemical staining. Statistical analysis was performed using mixed effects models. Results: We confirmed our hypothesis that differences in histopathological scores, mucosal gene expression and number of MPO-positive cells of biopsies at various distances from ulcers exist. Biopsies from the edge of the ulcer yield a higher histopathological score, higher mRNA expression of pro-inflammatory genes and higher number of MPO-positive cells compared to biopsies procured next to the ulcer in the colon and ileum (p<0.005). Conclusion: The location for biopsy procurement which yields the highest histopathological score and highest mRNA expression of pro-inflammatory genes is the ulcer edge.

• Keywords: Crohn's disease, biopsy, histology, histological remission, transcriptomics