Abstract 1717: High NOXA (PMAIP1) transcript levels combined with a short-lived NOXA protein primes mantle cell lymphoma (MCL) cells for death by inhibition of the ubiquitin proteasome system

MCL is an aggressive type of non-Hodgkin lymphoma with poor prognosis and short survival. The genetic hallmark of MCL is the t(11;14) translocation leading to an aberrant expression of the oncogene cyclin D1. Several other deregulated pathways also contribute to the pathogenesis of MCL, including the DNA damage response-, B-cell receptor- and PI3K/mTOR pathway. Deregulated oncogenic pathways not only induce proliferation but also frequently lead to a constitutive stress signal. A common response to cellular stress is the transcriptional up-regulation of pro-apoptotic genes. By analyzing mRNA expression of these genes in MCL cells we found high levels of the pro-apoptotic Bcl-2 family member NOXA (PMAIP1). NOXA transcript was significantly higher in MCL cell lines as well as in samples from MCL patients when compared to other cancer cell lines and PBMCs. To analyze if these high transcript levels depend on activation of oncogenic pathways, we treated the MCL cells with a panel of inhibitors of different signaling pathways and found that the inhibition of the PI3K/mTOR pathway led to a significant reduction of NOXA mRNA levels. These results indicate that this signaling axis is not only acting pro-survival but also mediates up-regulation of NOXA mRNA. In contrast to the high transcript levels, NOXA protein is low in MCL cells. We found that NOXA protein is unstable with half-life times of 15-30 min. Inhibitors of the ubiquitin proteasome system (UPS) such as bortezomib, MG-132 and the cullin-ubiquitin ligase inhibitor MLN4924 stabilized NOXA and led to a strong accumulation of the protein. This was accompanied by a rapid induction of cell death. Interestingly, similar effects could be observed using the fatty acid synthase inhibitor orlistat indicating that fatty acid metabolism is also involved in the UPS. Reducing the high NOXA mRNA levels by treating the cells with the PI3K/mTOR dual-inhibitor Bez235 or using RNA interference prior to treatment with UPS inhibitors significantly reduced cell death and NOXA protein accumulation. These results indicate that the high NOXA mRNA levels are essential for the response of MCL cells to proteasomal inhibitors. In summary, our results show that MCL have a constitutive signal mediated by the PI3K/mTOR pathway resulting in a high expression of NOXA mRNA. The cells survive by rapidly degrading the NOXA protein. Therefore, an effective strategy to kill MCL cells is to target the high NOXA protein turnover in these cells