ANALYSIS OF THE EFFECT OF NOVEL PKC ISOZYMES ON AUTOPHAGY AND UTILIZATION OF NANOPARTICLES AS A NEW THERAPEUTIC APPROACH FOR GAUCHER DISEASE

Özlem Yedier
Molecular Biology, Genetics and Bioengineering, MSc Thesis, 2017

Thesis Jury

Assoc. Prof. Devrim Gözüaçık (Thesis Advisor), Assoc. Prof. Özlem Kutlu (Thesis Co-advisor), Prof. Dr. Serap Dökmeci, Assoc. Prof. Funda Yağcı Acar, Assist. Prof. Murat Kaya Yapıcı

Date & Time: 28.07.2017 – 11 AM

Place: L055

Keywords : Autophagy, protein kinase C, shRNA screening, Gaucher disease, nanoparticle

Abstract

Autophagy is a mechanism that degrades long-lived proteins and non-functional organelles to sustain cellular homeostasis. Defects or abnormalities in this mechanism are associated with various diseases including cancer and neurodegeneration. Protein Kinase C (PKC) isozymes are a large family of serine-threonine kinases that have many roles inside the cell. Several studies showed the relations of PKC isozymes in autophagy pathway mostly in the induction step. However, in most of the cases, complete mechanisms or the specific isozyme responsible from the particular effect was not identified. In this study, the effect of active PKC isozymes on autophagy was analyzed and candidate PKC isozymes, that are important for the modulation of autophagy by PMA were identified. Moreover, lentiviral transduction method was established in order to use a lentiviral shRNA library for screening the target molecules functioning as effectors between PKC isozymes and autophagy.

Defects in lysosomal enzymes are the main cause of malfunctional autophagy and thus lysosomal storage diseases. Gaucher Disease is the most common lysosomal storage disease around the world that results from mutations in GBA1 gene which expresses glucocerebrosidase enzyme. In this study, acidic nanoparticles were utilized as an alternative methodology to functionalize impaired autophagic flux.