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- Article (55)
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- English (57) (remove)
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Institute
- Georg-Speyer-Haus (57) (remove)
Eißmann, Moritz ; Schwamb, Bettina ; Melzer, Inga Maria ; Moser, Julia ; Siele, Dagmar ; Köhl, Ulrike ; Rieker, Ralf Joachim ; Wachter, David Lukas ; Agaimy, Abbas ; Herpel, Esther ; Baumgarten, Peter ; Mittelbronn, Michel Guy André ; Rakel, Stefanie ; Kögel, Donat ; Böhm, Stefanie ; Gutschner, Tony ; Diederichs, Sven ; Zörnig, Martin
Yeast cells can be killed upon expression of pro-apoptotic mammalian proteins. We have established a functional yeast survival screen that was used to isolate novel human anti-apoptotic genes overexpressed in treatment-resistant tumors. The screening of three different cDNA libraries prepared from metastatic melanoma, glioblastomas and leukemic blasts allowed for the identification of many yeast cell death-repressing cDNAs, including 28% of genes that are already known to inhibit apoptosis, 35% of genes upregulated in at least one tumor entity and 16% of genes described as both anti-apoptotic in function and upregulated in tumors. These results confirm the great potential of this screening tool to identify novel anti-apoptotic and tumor-relevant molecules. Three of the isolated candidate genes were further analyzed regarding their anti-apoptotic function in cell culture and their potential as a therapeutic target for molecular therapy. PAICS, an enzyme required for de novo purine biosynthesis, the long non-coding RNA MALAT1 and the MAST2 kinase are overexpressed in certain tumor entities and capable of suppressing apoptosis in human cells. Using a subcutaneous xenograft mouse model, we also demonstrated that glioblastoma tumor growth requires MAST2 expression. An additional advantage of the yeast survival screen is its universal applicability. By using various inducible pro-apoptotic killer proteins and screening the appropriate cDNA library prepared from normal or pathologic tissue of interest, the survival screen can be used to identify apoptosis inhibitors in many different systems.