In vitro vēža šūnu modeļsistēmas izstrāde molekulāri mērķētu radionuklīdu testēšanai
Date
2019
Authors
Rubena, Elza Marija
Journal Title
Journal ISSN
Volume Title
Publisher
Latvijas Universitāte
Abstract
Prostatas vēzis ir ceturtais biežāk diagnosticētais vēzis Eiropā, 2018. gadā un tas ir kļuvis par visbiežāk sastopamo vēzi vīriešu vidū. Nozīmīgi klīnisko pētījumu rezultāti prostatas vēža diagnostikā un terapijā ir sasniegti ar radioaktīvi iezīmētu PSMA ligandu 68Ga-PSMA – diagnostikai un 177Lu-PSMA – terapijai. Neiroendokrīnie audzēji ir neviendabīga agresīvu audzēju grupa, kam raksturīgi dažādi un bieži nespecifiski simptomi, kas apgrūtina precīzu audzēja primāro diagnostiku un saslimšanas stadijas precizēšanu. Pašlaik šo audzēju diagnostikā un ārstēšanā izmanto ar SSTR2 ligandiem (somatostatīna analogiem) konjugētus radionuklīdus. Šī darba mērķis bija izveidot uz dažādām šūnu līnijām balstītu in vitro modeļsistēmu, kas varētu tikt izmantota molekulāri mērķētu radionuklīdu īpašību raksturošanā un to funkcionālās ietekmes izvērtēšanā. Šī darba ietvaros tika kultivētas sešas dažādas šūnu līnijas, kurās imūncitoķīmiski tika raksturota mērķreceptoru PSMA un SSTR2 ekspresija, un iegūtie rezultāti tālāk validēti molekulāri mērķētu radionuklīdu saistīšanās reakcijās un analizēti ar Furjē transformācijas infrasarkanās (FTIS) spektroskopijas metodi. Darba rezultātā izvēlētajās šūnu līnijās tika noteikta SSTR2 un PSMA ekspresija, kas tika validēta, izvērtējos radionuklīdu saistīšanās reakciju. Balsoties uz šiem rezulatātiem, tika izveidoti prostatas vēža un neiroendokrīno šūnu līniju paneļi, kas ietvēra vēža šūnas ar izteiktu un vāji izteiktu mērķreceptoru ekspresiju, kā arī normālu šūnu kontroli. Prostatas vēža šūnu panelī tika iekļautas PC šūnu līnijas LNCaP (PSMA++), PC3 (PSMA+/-) un dermālie fibroblasti Hs68; neiroendokrīno audzēju panelī – NET šūnu līnijas AR42J (SSTR2++), NCI-H69 (SSTR2+), CorL23 (SSTR2-) un Hs68. Izmantojot prostatas vēža modeļsistēmu, tika parādīta radionuklīda 177Lu-PSMA I&T iespējamā ietekme uz šūnu biomolekulāro profilu izmaiņām. Izstrādātās modeļsistēmas paver iespēju tālākiem jauniem molekulāri mērķētu radionuklīdu funkcionālās ietekmes pētījumiem šūnu līmenī.
Prostate cancer is the fourth most commonly diagnosed cancer in Europe in 2018 and has become the most common cancer among men. Significant results of clinical trials in diagnostics and treatment of prostate cancer have been achieved with radiolabelled PSMA ligands for 68Ga-PSMA - diagnostics and 177Lu-PSMA - therapy. Neuroendocrine tumours are a heterogeneous group of aggressive tumours with different and often non-specific symptoms that makes it difficult to pinpoint the diagnosis of primary tumour and the stage of disease. Currently, these radionuclides conjugated to SSTR2 ligands (somatostatin analogues) are used in the diagnosis and treatment of these tumours. The aim of this work was to create an in vitro model system based on different cell lines that could be used to characterize molecularly targeted radionuclides and to assess their functional effects. Six different cell lines were cultivated within this work, in which the expression of the target receptors PSMA and SSTR2 were testes by immunocytochemical method and the results obtained were further validated by molecularly targeted radionuclide binding reactions. Results were analysed by Fourier transform infrared (FTIR) spectroscopy. As a result of the work, the expression of SSTR2 and PSMA, which was validated, was determined in the selected cell lines and evaluated for the radionuclide binding reaction. Based on these results, panels of prostate cancer and neuroendocrine cell lines were created, including cancer cells with pronounced and weakly expressed target receptors as well as normal cell control. The prostate cancer cell panel included PC cell lines LNCaP (PSMA ++), PC3 (PSMA +/-) and dermal fibroblasts Hs68; neuroendocrine tumour panel - NET cell lines AR42J (SSTR2 ++), NCI-H69 (SSTR2 +), CorL23 (SSTR2-) and Hs68. Using the prostate cancer model system, the potential effect of radionuclide 177Lu-PSMA I&T on changes in cellular biomolecular profiles was demonstrated. The developed model systems provide an opportunity for further research on the functional effects of molecularly targeted radionuclides at the cellular level.
Prostate cancer is the fourth most commonly diagnosed cancer in Europe in 2018 and has become the most common cancer among men. Significant results of clinical trials in diagnostics and treatment of prostate cancer have been achieved with radiolabelled PSMA ligands for 68Ga-PSMA - diagnostics and 177Lu-PSMA - therapy. Neuroendocrine tumours are a heterogeneous group of aggressive tumours with different and often non-specific symptoms that makes it difficult to pinpoint the diagnosis of primary tumour and the stage of disease. Currently, these radionuclides conjugated to SSTR2 ligands (somatostatin analogues) are used in the diagnosis and treatment of these tumours. The aim of this work was to create an in vitro model system based on different cell lines that could be used to characterize molecularly targeted radionuclides and to assess their functional effects. Six different cell lines were cultivated within this work, in which the expression of the target receptors PSMA and SSTR2 were testes by immunocytochemical method and the results obtained were further validated by molecularly targeted radionuclide binding reactions. Results were analysed by Fourier transform infrared (FTIR) spectroscopy. As a result of the work, the expression of SSTR2 and PSMA, which was validated, was determined in the selected cell lines and evaluated for the radionuclide binding reaction. Based on these results, panels of prostate cancer and neuroendocrine cell lines were created, including cancer cells with pronounced and weakly expressed target receptors as well as normal cell control. The prostate cancer cell panel included PC cell lines LNCaP (PSMA ++), PC3 (PSMA +/-) and dermal fibroblasts Hs68; neuroendocrine tumour panel - NET cell lines AR42J (SSTR2 ++), NCI-H69 (SSTR2 +), CorL23 (SSTR2-) and Hs68. Using the prostate cancer model system, the potential effect of radionuclide 177Lu-PSMA I&T on changes in cellular biomolecular profiles was demonstrated. The developed model systems provide an opportunity for further research on the functional effects of molecularly targeted radionuclides at the cellular level.
Description
Keywords
Bioloģija , Prostatas vēzis , neiroendokrīnais vēzis , in vitro modeļsistēma , SSTR2 un PSMA , 68Ga un 177Lu radionuklīdi