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Post-transcriptional regulation of glutaminase enzyme by HuR and its relationship with high glutaminolytic levels in tumors

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Author(s):
Douglas Adamóski
Total Authors: 1
Document type: Doctoral Thesis
Institution: Universidade Estadual de Campinas, Instituto de Biologia
Defense date:
Advisor: Sandra Martha Gomes Dias
Abstract

TCA cycle anaplerosis by glutamine, an important energetic and biosynthetic source for rapidly dividing cells, is impaired by glutaminase inhibition. Glutaminase inhibition by CB-839 is currently under clinical trials for treating multiple solid and haematological tumors. HuR (Human antigen R) is a protein encoded by the ELAVL1 gene. HuR contains three RNA-binding domains and binds cis-acting AU-rich elements, affecting mRNA stability and splicing. Previous findings have linked HuR to GLS mRNA stabilization in renal acidosis, and its ortholog, HuD, to GLS splicing in neuronal cells. We then hypothesized that HuR was an important glutaminase regulator in cancer. We first inquired a TCGA pan-cancer cohort and found that patients with higher ELAVL1 expression had increased risk of tumor recidive (Cox HR of 1.82 for breast cancer). Moreover, by using publicly available RNA-Seq, RIP-Seq and PAR-CLIP-Seq data we showed that HuR binds to GLS intron 14 and controls GLS splicing, leading to kidney type glutaminase (KGA) choice over glutaminase C (GAC). Using breast and prostate cancer and HEK293 (with a CRISPR/Cas9 knock in) cell lines we further confirmed that HuR increases KGA by affecting splicing and mRNA stability. Lentiviral ELAVL1 silencing impaired breast cancer cell¿s proliferation, migration, invasion and increased glutamine addiction in vitro. Treating silenced ELAVL1 breast cancer cells with CB839 further impaired proliferation and invasion. We proposed HuR as a prognostic factor in breast cancer, with special impact on glutamine metabolism through splicing regulation. In this thesis, we also present seven published articles on which I participated as a co-author, being four as co-first author (AU)

FAPESP's process: 14/17820-3 - Post-transcriptional regulation of glutaminase enzyme by HuR and its relationship with high glutaminolytic levels in tumors
Grantee:Douglas Adamoski Meira
Support type: Scholarships in Brazil - Doctorate (Direct)