Uvođenje metode za mjerenje aktivnosti lizosomske kisele lipaze u uzorku suhe kapi krvi na filtar papiru

master's thesis

• Title (english): Introduction of method for measuring the activity of lysosomal acid lipase in dried blood filter paper samples
• Author: Vedrana Ivanković
• Mentor: Ksenija Fumić (mentor)
• Mentor: Marija Grdić Rajković (mentor)
• Committee member: Marija Grdić Rajković
• Committee member: Ksenija Fumić
• Committee member: Petra Turčić
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; Zagreb
• Defense date and country: 2015-07-13, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Wolmanova bolest i bolest nakupljanja kolesterolskih estera su autosomno recesivne nasljedne bolesti koje
karakterizira manjak enzima lizosomske kisele lipaze. Lizosomska kisela lipaza je hidrolaza koja je ključna za
razgradnju lipoproteina plazme u lizosomima. Zbog prirođenog manjka lizosomske kisele lipaze dolazi do nakupljanja
kolesterolskih estera i triglicerida u tkivima koja najviše sudjeluju u endocitozi posredovanoj receptorima, a to su
stanice jetre, crijeva, nadbubrežnih žlijezda, limfnih čvorova i općenito stanice monocitno-makrofagnog sustava.
Novije klinčke studije pokazuju ohrabrujuće liječenje nadomjesnom terapijom rekombinantnim enzimom sebelipaza
alfa. Cilj rada je uvođenje metode probira za mjerenje lizosomske kisele lipaze iz uzorka suhe kapi krvi na filtar
papiru. Mjerenjem ukupne aktivnosti lipaza i mjerenjem aktivnosti lipaza u prisutnosti Lalistata 2, lizosomska kisela
lipaza može se dokazati u uzorku suhe kapi krvi. Rezultati dobiveni mjerenjima nakon uspostavljanja optimalnih
uvjeta pokazuju dobro slaganje s literaturnim podacima. Dobiveni rezultati naše „in house“ metode probira pokazali
su da se metoda može koristiti za razlikovanje zdravih osoba od pacijenata s manjkom lizosomske kisele lipaze.
Metoda nije pouzdana za razlikovanje heterozigota od zdravih osoba. Da bi se ova „in house“ metoda mogla koristiti u
kliničkom laboratoriju kao metoda probira pri sumnji na manjak lizosomske kisele lipaze, potrebno je napraviti
validaciju metode prema odgovarajućem protokolu za uzorke suhe kapi krvi na filtar papiru. Validirana metoda
probira iz uzorka suhe kapi krvi na filtar papiru omogućit će veću dostupnost dijagnostike manjka lizosomske kisele
lipaze i pravovremeno uvođenje enzimskog nadomjesnog liječenja do sada nelječivih nasljednih metaboličkih
poremećaja (Wolmanove bolesti i bolesti nakupljanja kolesterolskih estera).

Abstract (english)

Wolman disease and Cholesterol ester storage disease are autosoma lrecessive hereditary diseases characterized by a
deficiency of the enzyme lysosomal acid lipase. Lysosomal acid lipase is a hydrolase which is essential for
degradation of plasma lipoproteins in the lysosomes. Because of the inherent lack of lysosomal acid lipase leads to
accumulation of cholesterol esters and triglycerides in tissues that most participate in mediated endocytosis receptors
and these are the cells of the liver, spleen, adrenal glands, lymphnodes and cells in general monocyte macrophage
system. Recent clinical studies show encouraging treatment by recombinant replacement therapy with sebelipaza alpha.
Objective of this paper is to introduce screening methods for the measurement of lysosomal acid lipase in dried blood
filter paper samples. By measuring total lipase activity and lipase activity in the presence of Lalistat 2, lysosomal acid
lipase can be determined in dried blood samples. Results obtained by measurement after establishment of optimal conditions show good agreement with the literature data. The results of our „in house“ screening method have shown that the method can be used to distinguish healthy individuals from patients with a lack of lysosomal acid lipase. Method is not reliable for distinguishing heterozygotes from healthy individuals. Validation of the „in house“ method according to the appropriate protocol for dried blood filter paper samples is necessary when it is used in clinical laboratories as a method in suspected lack lysosome acid lipase.Validated method of selecting from dried blood filter paper samples will enable greater availability of diagnosing a missing lysosomal acid lipase and the timely introduction of enzyme replacement therapy for so far incurable hereditary metabolic disorders (Wolman disease and Cholesterol ester storage disease).

• Keywords: Wolmanova bolest; bolest nakupljanja kolesterolskih estera; lizosomska kisela lipaza; uzorak suhe kapi krvi
• Keywords (english): Wolman disease; Cholesterol ester storage disease; lysosomal acid lipase; dried blood spots
• Language: croatian
• URN:NBN: urn:nbn:hr:163:597354
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2016-01-20 13:21:55