Naslov
Function of the long noncoding RNA, WNT1-inducible signaling pathway protein 1-overlapping transcript 2 (WISP1-OT2) in human renal cells

Autori
Mirjana Polović, Gerald Schwerdt, Michael Gekle

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
96th Annual Meeting of the German Physiological Society

Mjesto i datum
Greifswald, Njemačka, 16.- 18.3. 2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
OTA, ochratoxin A, lncRNA, WISP1

Sažetak
Poster A01-2: Background: The alternative transcript WISP1-OT2 (WNT1 inducible signaling pathway protein 1 overlapping transcript 2) was discovered upon treatment of renal proximal tubule epithelial cells (RPTEC) with the ubiquitous mycotoxin, ochratoxin A (OTA). WISP1-OT2 is a long noncoding RNA, 2922 nucleotides long, and transcribed from the end of WISP1 gene. We were able to detect WISP1-OT2 in healthy renal tissue and we observed an upregulation in clear cell renal cell carcinoma. WISP1-OT2 is located predominantly in the nuclear fraction ; however, its function is unknown. Aims: (i) Determine the orientation of WISP1-OT2 ; (ii) investigate possible functional link between long noncoding RNA and mRNA of WISP1 gene ; (iii) evaluate the influence of WISP1-OT2 knockdown on the cell function. Methods: Orientation of WISP1-OT2 was investigated by gene-specific reverse transcription and subsequent PCR. To test an inhibitory effect of WISP1-OT2 on WISP1 mRNA, HEK293T cells were treated either with PMA (Phorbol myristate acetate) or OTA alone, or in combination, in the presence or absence of antisense LNA™ GapmeRs, i.e. hybrid DNA:RNA oligonucleotides that specifically enter the nucleus and degrade target RNA. Subsequently, WISP1-OT2 and WISP1 mRNA expression was detected by RT-PCR and ddPCR. To investigate the importance of WISP1-OT2 for gene expression, RNA-seq data were generated in the absence and presence of specific antisense oligonucleotides against WISP1-OT2. Furthermore, we determined media acidification, HCO - consumption, glucose consumption, lactate production, caspase-III activation and LDH release. Results: WISP1-OT2 is antisense-oriented long noncoding RNA. PMA induces specifically WISP1 mRNA, whereas OTA induces specifically WISP1-OT2. In the presence of PMA and OTA, WISP1 mRNA is significantly reduced when compared to PMA effect alone. This inhibitory effect of OTA on the WISP1 mRNA is not mediated by WISP1-OT2. Furthermore, preliminary experiments indicate that WISP1-OT2 modulates cellular energy metabolism and acid-base balance. Conclusions: (i) WISP1-OT2 is the long noncoding RNA, transcribed in antisense orientation relative to WISP1 gene ; (ii) WISP1-OT2 does not influence the expression of WISP1 mRNA ; (iii) WISP1-OT2 interferes with cellular energy metabolism.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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