Individualizovana terapija - uloga proteinskih i genetskih varijanti tiopurin S-metiltransferaze
Individualized therapy: Role of thiopurine S-methyltransferase protein and genetic variants
Apstrakt
Tiopurin S-metiltransferaza (TPMT: EC 2.1.1.67) jeste enzim koji metaboliše imunosupresivne tiopurinske lekove, koji se koriste za lečenje autoimunih bolesti, malignih oboljenja i u transplantacionoj medicini. Aktivnost enzima TPMT kod pojedinih ljudi je izrazito smanjena ili povećana u odnosu na normalni nivo aktivnosti. Istraživanja strukture i biohemijskih karakteristika proteina TPMT su ukazala na postojanje određenih proteinskih varijanti koje imaju izmenjenu aktivnost. Otkriveni su polimorfizmi u genu za TPMT koji daju različite TPMT alozime. Smanjenoj aktivnosti enzima može doprineti i manja količina sintetisanog proteina, što zavisi i od transkripcione aktivnosti promotora gena za TPMT. Polimorfizmi u samom promotoru, kao što je promenjiv broj tandemskih ponovaka (VNTR), mogu da modulišu transkripciju. Primena tiopurinskih lekova kod pacijenata sa određenim genetskim varijantama TPMT izaziva tešku hematološku toksičnost. Da bi se toksičnost izbegla, terapija se modifikuje u skl...adu sa genotipom TPMT (farmakogenetika). Mi smo izučavali polimorfizme u egzonima i regulatornim elementima (promotor) gena za TPMT koji dovode do promene aktivnosti enzima TPMT u srpskoj populaciji. Koristili smo metodologiju baziranu na PCR i DNK sekvenciranje za detekciju genetskih varijanti TPMT. Pokazali smo da su u našoj populaciji prisutne genetske varijante u egzonima koje ukupno daju 7,5% varijantnih alozima TPMT koji imaju smanjenu enzimsku aktivnost. Terapija za pacijente koji imaju ove farmakogenetičke markere je modifikovana, što je doprinelo uspešnijem lečenju. Funkcionalnim esejima in vitro smo pokazali da aktivnost promotora gena za TPMT, a samim tim i količina sintetisanog enzima TPMT, zavisi od arhitekture (broja i tipa) VNTR u promotoru. Promotor gena za TPMT specifično odgovara na tretman ćelija K562 tiopurinom zavisno od tipa VNTR. Izučavanje interakcija DNK i proteina je otkrilo da transkripcioni faktori Sp1 i Sp3 interaguju sa VNTR. Naša istraživanja ukazuju na to da bi region VNTR u promotoru gena za TPMT mogao postati novi farmakogenetički marker od kliničkog značaja za individualizaciju tiopurinske terapije.
Thiopurine S-methyltransferase (TPMT: EC 2.1.1.67) is an enzyme that metabolizes immunosuppressive thiopurine medications, used in the treatment of autoimmune diseases, cancer and in transplantation medicine. In some individuals, TPMT enzyme activity is significantly increased or decreased compared to the normal TPMT activity level. Structural and biochemical analyses of the TPMT protein revealed the existence of certain protein variants with altered activity. It has been shown that certain TPMT gene polymorphisms exist, that define different TPMT allozymes. Decreased TPMT enzyme activity can also be a consequence of lower protein synthesis, which depends on the promoter transcription activity. Promoter polymorphisms, such as variable number of tandem repeats (VNTR), can modulate the transcription. Administering thiopurine drugs in patients with certain genetic TPMT variants leads to severe hematologic toxicity. To avoid toxicity, therapy is being modified according to the TPMT genotyp...e (pharmacogenetics). We investigated the polymorphisms in exons and regulatory elements (promoter) of the TPMT gene which affect TPMT enzyme activity in the Serbian population. We used PCR-based methodology and sequencing in the detection of genetic variants on TPMT gene. We showed that genetic variants in exons account for 7.5% of all TPMT variants with decreased enzyme activity. The therapy for patients with these pharmacogenetic markers was modified, which contributed to the efficiency of treatment. Functional assays in vitro showed that the TPMT promoter activity and, therefore, the quantity of TPMT protein synthesized, depended on the architecture of VNTRs (i.e. number and type) in the promoter. Promoter of the TPMT gene specifically responds to mercaptopurine treatment of K562 cells in a VNTR-dependent manner. Study of DNA-protein interactions revealed that Sp1 and Sp3 transcription factors interact with VNTRs. Our research pointed out that the VNTR promoter region of the TPMT gene could become a new pharmacogenetic marker, clinically significant for the individualization of thiopurine therapy.
Ključne reči:
VNTR u promotoru gena za TPMT / polimorfizmi u genu za TPMT / farmakogenetika / alozimi TPMT / VNTR in TPMT gene promoter / TPMT gene polymorphisms / TPMT allozyme / pharmacogeneticsIzvor:
Journal of Medical Biochemistry, 2010, 29, 3, 150-156Izdavač:
- Društvo medicinskih biohemičara Srbije, Beograd i Versita
Finansiranje / projekti:
- Strukturalni elementi genoma u modulaciji fenotipa (RS-MESTD-MPN2006-2010-143051)
DOI: 10.2478/v10011-010-0023-x
ISSN: 1452-8258
WoS: 000280477800004
Scopus: 2-s2.0-77955248330
Institucija/grupa
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Pavlović, Sonja AU - Zukić, Branka PY - 2010 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/457 AB - Tiopurin S-metiltransferaza (TPMT: EC 2.1.1.67) jeste enzim koji metaboliše imunosupresivne tiopurinske lekove, koji se koriste za lečenje autoimunih bolesti, malignih oboljenja i u transplantacionoj medicini. Aktivnost enzima TPMT kod pojedinih ljudi je izrazito smanjena ili povećana u odnosu na normalni nivo aktivnosti. Istraživanja strukture i biohemijskih karakteristika proteina TPMT su ukazala na postojanje određenih proteinskih varijanti koje imaju izmenjenu aktivnost. Otkriveni su polimorfizmi u genu za TPMT koji daju različite TPMT alozime. Smanjenoj aktivnosti enzima može doprineti i manja količina sintetisanog proteina, što zavisi i od transkripcione aktivnosti promotora gena za TPMT. Polimorfizmi u samom promotoru, kao što je promenjiv broj tandemskih ponovaka (VNTR), mogu da modulišu transkripciju. Primena tiopurinskih lekova kod pacijenata sa određenim genetskim varijantama TPMT izaziva tešku hematološku toksičnost. Da bi se toksičnost izbegla, terapija se modifikuje u skladu sa genotipom TPMT (farmakogenetika). Mi smo izučavali polimorfizme u egzonima i regulatornim elementima (promotor) gena za TPMT koji dovode do promene aktivnosti enzima TPMT u srpskoj populaciji. Koristili smo metodologiju baziranu na PCR i DNK sekvenciranje za detekciju genetskih varijanti TPMT. Pokazali smo da su u našoj populaciji prisutne genetske varijante u egzonima koje ukupno daju 7,5% varijantnih alozima TPMT koji imaju smanjenu enzimsku aktivnost. Terapija za pacijente koji imaju ove farmakogenetičke markere je modifikovana, što je doprinelo uspešnijem lečenju. Funkcionalnim esejima in vitro smo pokazali da aktivnost promotora gena za TPMT, a samim tim i količina sintetisanog enzima TPMT, zavisi od arhitekture (broja i tipa) VNTR u promotoru. Promotor gena za TPMT specifično odgovara na tretman ćelija K562 tiopurinom zavisno od tipa VNTR. Izučavanje interakcija DNK i proteina je otkrilo da transkripcioni faktori Sp1 i Sp3 interaguju sa VNTR. Naša istraživanja ukazuju na to da bi region VNTR u promotoru gena za TPMT mogao postati novi farmakogenetički marker od kliničkog značaja za individualizaciju tiopurinske terapije. AB - Thiopurine S-methyltransferase (TPMT: EC 2.1.1.67) is an enzyme that metabolizes immunosuppressive thiopurine medications, used in the treatment of autoimmune diseases, cancer and in transplantation medicine. In some individuals, TPMT enzyme activity is significantly increased or decreased compared to the normal TPMT activity level. Structural and biochemical analyses of the TPMT protein revealed the existence of certain protein variants with altered activity. It has been shown that certain TPMT gene polymorphisms exist, that define different TPMT allozymes. Decreased TPMT enzyme activity can also be a consequence of lower protein synthesis, which depends on the promoter transcription activity. Promoter polymorphisms, such as variable number of tandem repeats (VNTR), can modulate the transcription. Administering thiopurine drugs in patients with certain genetic TPMT variants leads to severe hematologic toxicity. To avoid toxicity, therapy is being modified according to the TPMT genotype (pharmacogenetics). We investigated the polymorphisms in exons and regulatory elements (promoter) of the TPMT gene which affect TPMT enzyme activity in the Serbian population. We used PCR-based methodology and sequencing in the detection of genetic variants on TPMT gene. We showed that genetic variants in exons account for 7.5% of all TPMT variants with decreased enzyme activity. The therapy for patients with these pharmacogenetic markers was modified, which contributed to the efficiency of treatment. Functional assays in vitro showed that the TPMT promoter activity and, therefore, the quantity of TPMT protein synthesized, depended on the architecture of VNTRs (i.e. number and type) in the promoter. Promoter of the TPMT gene specifically responds to mercaptopurine treatment of K562 cells in a VNTR-dependent manner. Study of DNA-protein interactions revealed that Sp1 and Sp3 transcription factors interact with VNTRs. Our research pointed out that the VNTR promoter region of the TPMT gene could become a new pharmacogenetic marker, clinically significant for the individualization of thiopurine therapy. PB - Društvo medicinskih biohemičara Srbije, Beograd i Versita T2 - Journal of Medical Biochemistry T1 - Individualizovana terapija - uloga proteinskih i genetskih varijanti tiopurin S-metiltransferaze T1 - Individualized therapy: Role of thiopurine S-methyltransferase protein and genetic variants EP - 156 IS - 3 SP - 150 VL - 29 DO - 10.2478/v10011-010-0023-x ER -
@article{ author = "Pavlović, Sonja and Zukić, Branka", year = "2010", abstract = "Tiopurin S-metiltransferaza (TPMT: EC 2.1.1.67) jeste enzim koji metaboliše imunosupresivne tiopurinske lekove, koji se koriste za lečenje autoimunih bolesti, malignih oboljenja i u transplantacionoj medicini. Aktivnost enzima TPMT kod pojedinih ljudi je izrazito smanjena ili povećana u odnosu na normalni nivo aktivnosti. Istraživanja strukture i biohemijskih karakteristika proteina TPMT su ukazala na postojanje određenih proteinskih varijanti koje imaju izmenjenu aktivnost. Otkriveni su polimorfizmi u genu za TPMT koji daju različite TPMT alozime. Smanjenoj aktivnosti enzima može doprineti i manja količina sintetisanog proteina, što zavisi i od transkripcione aktivnosti promotora gena za TPMT. Polimorfizmi u samom promotoru, kao što je promenjiv broj tandemskih ponovaka (VNTR), mogu da modulišu transkripciju. Primena tiopurinskih lekova kod pacijenata sa određenim genetskim varijantama TPMT izaziva tešku hematološku toksičnost. Da bi se toksičnost izbegla, terapija se modifikuje u skladu sa genotipom TPMT (farmakogenetika). Mi smo izučavali polimorfizme u egzonima i regulatornim elementima (promotor) gena za TPMT koji dovode do promene aktivnosti enzima TPMT u srpskoj populaciji. Koristili smo metodologiju baziranu na PCR i DNK sekvenciranje za detekciju genetskih varijanti TPMT. Pokazali smo da su u našoj populaciji prisutne genetske varijante u egzonima koje ukupno daju 7,5% varijantnih alozima TPMT koji imaju smanjenu enzimsku aktivnost. Terapija za pacijente koji imaju ove farmakogenetičke markere je modifikovana, što je doprinelo uspešnijem lečenju. Funkcionalnim esejima in vitro smo pokazali da aktivnost promotora gena za TPMT, a samim tim i količina sintetisanog enzima TPMT, zavisi od arhitekture (broja i tipa) VNTR u promotoru. Promotor gena za TPMT specifično odgovara na tretman ćelija K562 tiopurinom zavisno od tipa VNTR. Izučavanje interakcija DNK i proteina je otkrilo da transkripcioni faktori Sp1 i Sp3 interaguju sa VNTR. Naša istraživanja ukazuju na to da bi region VNTR u promotoru gena za TPMT mogao postati novi farmakogenetički marker od kliničkog značaja za individualizaciju tiopurinske terapije., Thiopurine S-methyltransferase (TPMT: EC 2.1.1.67) is an enzyme that metabolizes immunosuppressive thiopurine medications, used in the treatment of autoimmune diseases, cancer and in transplantation medicine. In some individuals, TPMT enzyme activity is significantly increased or decreased compared to the normal TPMT activity level. Structural and biochemical analyses of the TPMT protein revealed the existence of certain protein variants with altered activity. It has been shown that certain TPMT gene polymorphisms exist, that define different TPMT allozymes. Decreased TPMT enzyme activity can also be a consequence of lower protein synthesis, which depends on the promoter transcription activity. Promoter polymorphisms, such as variable number of tandem repeats (VNTR), can modulate the transcription. Administering thiopurine drugs in patients with certain genetic TPMT variants leads to severe hematologic toxicity. To avoid toxicity, therapy is being modified according to the TPMT genotype (pharmacogenetics). We investigated the polymorphisms in exons and regulatory elements (promoter) of the TPMT gene which affect TPMT enzyme activity in the Serbian population. We used PCR-based methodology and sequencing in the detection of genetic variants on TPMT gene. We showed that genetic variants in exons account for 7.5% of all TPMT variants with decreased enzyme activity. The therapy for patients with these pharmacogenetic markers was modified, which contributed to the efficiency of treatment. Functional assays in vitro showed that the TPMT promoter activity and, therefore, the quantity of TPMT protein synthesized, depended on the architecture of VNTRs (i.e. number and type) in the promoter. Promoter of the TPMT gene specifically responds to mercaptopurine treatment of K562 cells in a VNTR-dependent manner. Study of DNA-protein interactions revealed that Sp1 and Sp3 transcription factors interact with VNTRs. Our research pointed out that the VNTR promoter region of the TPMT gene could become a new pharmacogenetic marker, clinically significant for the individualization of thiopurine therapy.", publisher = "Društvo medicinskih biohemičara Srbije, Beograd i Versita", journal = "Journal of Medical Biochemistry", title = "Individualizovana terapija - uloga proteinskih i genetskih varijanti tiopurin S-metiltransferaze, Individualized therapy: Role of thiopurine S-methyltransferase protein and genetic variants", pages = "156-150", number = "3", volume = "29", doi = "10.2478/v10011-010-0023-x" }
Pavlović, S.,& Zukić, B.. (2010). Individualizovana terapija - uloga proteinskih i genetskih varijanti tiopurin S-metiltransferaze. in Journal of Medical Biochemistry Društvo medicinskih biohemičara Srbije, Beograd i Versita., 29(3), 150-156. https://doi.org/10.2478/v10011-010-0023-x
Pavlović S, Zukić B. Individualizovana terapija - uloga proteinskih i genetskih varijanti tiopurin S-metiltransferaze. in Journal of Medical Biochemistry. 2010;29(3):150-156. doi:10.2478/v10011-010-0023-x .
Pavlović, Sonja, Zukić, Branka, "Individualizovana terapija - uloga proteinskih i genetskih varijanti tiopurin S-metiltransferaze" in Journal of Medical Biochemistry, 29, no. 3 (2010):150-156, https://doi.org/10.2478/v10011-010-0023-x . .