20.500.12556/RUL-186661 Vzpostavitev tumorske komponente singenetskega modela mišjega B-celičnega limfoma in molekularna potrditev urejanja gena Cd19 v celični liniji A20 Establishment of the tumour component of a syngeneic murine B-cell lymphoma model and molecular confirmation of Cd19 gene editing in the A20 cell line Rak je po svetu med tremi glavnimi vzroki prezgodnje smrti. Limfomi so skupina neoplazem, ki izvirajo iz limfocitov B ali limfocitov T perifernih limfatičnih organov, pri čemer 85 do 90 % limfomov izvira iz limfocitov B. Namen magistrskega dela je bil vzpostaviti in ovrednotiti tumorsko komponento singenetskega modela B-celičnega limfoma z uporabo celičnih populacij A20_CD19+ in A20_CD19_KO ter molekularno ovrednotiti modifikacijo gena Cd19 v celicah A20_CD19_KO. Celična populacija A20_CD19+ predstavlja mišjo B-celično limfomsko linijo z ohranjenim izražanjem površinskega antigena CD19, medtem ko je celična populacija A20_CD19_KO genetsko spremenjena tako, da ima zmanjšano oziroma odsotno izražanje proteina CD19. Samice mišje linije BALB/cAnNCrl smo subkutano injicirali s celicami A20_CD19+ ali A20_CD19_KO v odmerkih 0,5 × 10⁶, 1 × 10⁶ in 2 × 10⁶ celic. Spremljali smo pojavnost in rast tumorjev, telesno maso, zauživanje krme in vode ter hematološke in metabolne parametre krvi. Molekularno validacijo smo izvedli z določanjem zaporedja po Sangerjevi metodi, rezultate pa analizirali s programoma ICE in TIDE. Tumorji so se razvili pri vseh miših, injiciranih z populacijama celic A20, pri 11 % živali je prišlo do spontane regresije. Odmerek 1 × 10⁶ celic je bil povezan z najhitrejšo rastjo tumorjev. Obe populaciji sta pokazali primerljivo sposobnost tvorbe tumorjev, analiza zaporedij pa je potrdila prisotnost ciljnih sprememb na mestu urejanja gena Cd19 v celični populaciji A20_CD19_KO. Rezultati raziskave predstavljajo pomemben korak k vzpostavitvi singenetskega modela za raziskave imunoterapij. Vzpostavljena tumorska komponenta modela predstavlja uporabno osnovo za nadaljnje raziskave CD19-usmerjenih imunoterapij ter mehanizmov tumorskega pobega, povezanih z izgubo izražanja CD19. Cancer is one of the three leading causes of premature death worldwide. Lymphomas are a group of neoplasms originating from B or T lymphocytes of the peripheral lymphoid organs, with 85 to 90 % of lymphomas arising from B lymphocytes. The aim of this master’s thesis was to establish and evaluate the tumour component of a syngeneic B-cell lymphoma model using the A20_CD19+ and A20_CD19_KO cell populations, and to molecularly validate the modification of the Cd19 gene in the A20_CD19_KO cell population. The A20_CD19+ cells represent a murine B-cell lymphoma line with preserved expression of the surface antigen CD19, whereas the A20_CD19_KO cells have been genetically modified to exhibit reduced or absent expression of the CD19 protein. Female BALB/cAnNCrl mice were subcutaneously injected with A20_CD19+ or A20_CD19_KO cells at doses of 0,5 × 10⁶, 1 × 10⁶ and 2 × 10⁶ cells. Tumour incidence and growth, body weight, feed and water intake, as well as haematological and blood metabolic parameters, were monitored. Molecular validation was performed using Sanger sequencing, and the results were analysed using the ICE and TIDE software packages. Tumours developed in all mice injected with either A20 cell populations, although spontaneous tumour regression occurred in 11 % of the animals. The dose of 1 × 10⁶ cells was associated with the most rapid tumour growth. Both cell populations demonstrated comparable tumour-forming capacity, while sequence analysis confirmed the presence of the intended modifications at the Cd19 gene-editing site in the A20_CD19_KO cell population. The findings of this study represent an important step towards the establishment of a syngeneic model for immunotherapy research. The successfully established tumour component provides a valuable platform for further investigations of CD19-targeted immunotherapies and the mechanisms of tumour immune escape associated with the loss of CD19 expression. genetika imunologija medicina rak limfom singenetski model tumorji A20 B celice CAR-T CD19 laboratorijske miši genetics immunology medicine cancer lymphoma syngeneic model tumours A20 B cells CAR-T CD19 laboratory mice true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-09-04 07:17:32 2026-09-04 07:17:41 2026-09-05 04:33:49 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 2027-08-28 286860 60917.pdf 60917.pdf 1 9BA48579D8B0D032BBDFB9AD04A5359F ce722e737fe866181ef3f43325ca3ee5cea3c7e01469e3893d6a244de986b522 f40eb59f-a81f-11f1-8bc5-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245978 60918.pdf 60918.pdf 1 CDCA501B12409C38682B5AAD3107EF8A e8a1c76672953a668ba60b06d2e3cc9a06e9302b8603b3008d64e55afe7772d3 f411cd02-a81f-11f1-8bc5-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245979 Biotehniška fakulteta 0 0 0