Kloniranje liganda JAGGED1 i njegov utjecaj na proliferaciju limfocita

master's thesis

• Title (english): Cloning of the ligand JAGGED1 and its effect on the proliferation of lymphocytes
• Author: Maja Šetinc
• Mentor: Maja Matulić (mentor)
• Committee member: Maja Matulić (predsjednik povjerenstva)
• Committee member: Goran Kovačević (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: 2019-02-22, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Signalni put Notch igra ključnu ulogu u određivanju stanične sudbine u brojnim tkivima. Ovisno o vrsti stanica, aktivacija puta Notch može rezultirati diferencijacijom, proliferacijom ili apoptozom. Receptori Notch transmembranski su proteini s velikom ekstracelularnom domenom koja veže ligande, transmembranske proteine na susjednoj stanici. Prilikom vezanja liganda i receptora intracelularna domena receptora se proteolitički cijepa i putuje u jezgru, gdje se veže s transkripcijskim faktorom CSL i tako regulira transkripciju gena. Ligandi receptora Notch dijele se u dvije skupine – ligande "Delta-like" (DLL) i ligande Jagged. Signalni put Notch ima važnu ulogu u diferencijaciji krvnih stanica, a pogreške u regulaciji tog procesa mogu dovesti do transformacije stanica i nastanka leukemija. Cilj ovog istraživanja bio je analizirati utjecaj liganda JAGGED1 na proliferaciju limfocita i aktivaciju puta Notch. Konstruiran je ekspresijski plazmid sa sekvencijom liganda JAGGED1. Leukemijski B-limfociti uzgajani su u kokulturi s adherentnim stanicama koje eksprimiraju ovaj ligand. Pratila se proliferacija limfocita u kokulturi u odnosu na kulturu samih limfocita te onih tretiranih inhibitorom puta Notch, kako bi se utvrdilo utječe li aktivacija puta Notch ligandom JAGGED1 na biologiju limfocita.

Abstract (english)

Notch signaling pathway plays a crucial role in the determination of the cell fate in many tissues. Depending on the cell type, activation of Notch signaling can result in cell differentiation, proliferation or apoptosis. Notch receptors are transmembrane proteins with a large extracellular ligand-binding domain which binds to the ligand on the neighbouring cell. When a ligand binds to a receptor, the intracellular domain of the receptor is proteolytically cleaved and travels to the nucleus. There it binds CSL transcription factor and in that manner regulates gene transcription. Notch ligands belong to two families – "Delta-like" ligands (DLL) and Jagged. Notch signaling pathway plays an important role in the blood cell differentiation, and misregulations of that process could lead to malignant transformation and development of leukemia. The aim of this thesis was to analyse the effect of JAGGED1 ligand on the lymphocyte proliferation and activation of Notch signaling pathway. An expression plasmid with a coding sequence JAGGED1 was constructed, and leukemic B- lymphocytes were cocultured with adherent cell expressing JAGGED1 ligand. Proliferation of lymphocytes in coculture was analyzed in comparison with proliferation of lymphocytes grown alone, and lymphocytes treated with Notch inhibitor, to determine the effect of JAGGED1 ligand signaling on the lymphocyte biology.

• Keywords: Notch; signalni put; kokultura; leukemija; B-ALL
• Keywords (english): Notch; signaling pathway; coculture; leukemia; B-ALL
• Language: croatian
• URN:NBN: urn:nbn:hr:217:395678
• 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: 2021-02-22
• Created on: 2019-03-27 13:28:05