Istraživanje osjetljivosti stanične linije SU-DHL-5 s mutacijama gena KLHL6 na lijekove

master's thesis

• Title (english): Analysis of Drug Sensitivity Effects of KLHL6 Mutations in the SU-DHL-5 Cell Line
• Author: Luka Tandarić
• Mentor: Sirpa Leppa (mentor)
• Mentor: Petra Korać (mentor)
• Committee member: Petra Korać (predsjednik povjerenstva)
• Committee member: Antun Alegro (član povjerenstva)
• Committee member: Sofia Ana Blažević (član povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2019-10-16, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Difuzni velikostanični B-limfom (DLBCL, od eng. diffuse large B-cell lymphoma) najučestaliji je oblik ne-Hodgkinovog limfoma. Svjetska zdravstvena organizacija trenutno definira više tipova DLBCL-a, najučestaliji od kojih je DLBCL, neodređeni tip (NOS, od eng. not otherwise specified). Protein 6 sličan proteinu Kelch (KLHL6, od eng. kelch-like protein 6) uključen je u sazrijevanje B-limfocita i u otprilike 10% slučajeva DLBCL, NOS-a pronađena je njegova mutacija. Ciljevi ovog istraživanja bili su karakterizacija učinaka najučestalijih mutacija KLHL6 na odgovor na lijekove i posljedično otkrivanje staničnih putova u kojima KLHL6 sudjeluje. Stanice SU-DHL-5 (od eng. Stanford University diffuse histiocytic lymphoma 5), model DLBCL, NOS-a tipa GCB (od eng. germinal center B-cell-like), lentivirusno su tranducirane ekspresijskim konstruktima koji kodiraju proteine KLHL6 divljeg i mutiranog tipa. Radi sveobuhvatnog razotkrivanja učinaka najučestalijih mutacija na odgovor na lijekove, upotrebljen je panel od 528 lijekova, nakon čega su određene razlike u vijabilnosti između stanica SU-DHL-5 koje eksprimiraju nemutirane i mutirane gene za protein KLHL6. Početni rezultati probira sugerirali su da neke ispitane mutacije mijenjaju odgovor na određene skupine lijekova koji djeluju na isti stanični put, npr. na put PI3K/Akt. Međutim, nakon pokušaja potvrde rezultata, značajni utjecaji mutacija na odgovor na lijekove nisu uočeni, što dovodi postojanje učinaka ispitivanih mutacija KLHL6 u DLBCL-u u pitanje.

Abstract (english)

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma worldwide. Several types of DLBCL are currently recognized by the World Health Organization, with the most common type being DLBCL, NOS (not otherwise specified). Kelch-like protein 6 (KLHL6), a protein involved in B-lymphocyte maturation, has been found to carry mutations in about 10 % of DLBCL, NOS cases. The goals of this study were the characterization of the effects these mutations have on drug response, and consequent elucidation of cellular pathways KLHL6 is involved in. SU-DHL-5 cells, a GCB-type DLBCL, NOS model, were lentivirally-transduced with mutant and wild-type KLHL6 constructs to express the proteins. A panel of 528 drugs was used in order to comprehensively detect effects of the most common mutations on drug response, and differences in the viability between SU-DHL-5 cells expressing wild-type and mutant KLHL6 were determined. Preliminary drug screen results suggested that some mutations modify the response to certain groups of drugs affecting the same cellular pathway, e.g. the PI3K/Akt pathway. However, verification experiments were unable to reproduce the results, and no significant effects of KLHL6 mutations on drug response were observed, which puts the significance of the explored KLHL6 mutations in DLBCL into question.

• Keywords: transdukcija lentivirusima; DSRT; kelch-u sličan protein 6; neHodgkinov limfom
• Keywords (english): lentiviral transduction; DSRT; kelch-like protein 6; non-Hodgkin’s lymphoma
• Language: croatian
• URN:NBN: urn:nbn:hr:217:503841
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2021-10-16
• Created on: 2019-11-20 07:38:51