Tumor and Stem Cell Biology Combinatorial Regulation of Neuroblastoma Tumor Progression by N-Myc and Hypoxia Inducible Factor HIF-1a

In human neuroblastoma, amplification of theMYCN gene predicts poor prognosis and resistance to therapy. Because hypoxia contributes to aggressive tumor phenotypes, predominantly via two structurally related hypoxia inducible factors, HIF-1a and HIF-2a, we examined hypoxia responses inMYCN-amplified neuroblastoma cells. We demonstrate here that HIF-1a, but not HIF-2a, is preferentially expressed in both MYCN-amplified neuroblastoma cells and primary tumors in comparison to samples without MYCN amplification. Our results showed that interplay betweenN-Myc andHIF-1aplays critical roles in neuroblastoma. For example, high levels of N-Myc override HIF-1a inhibition of cell cycle progression, enabling continued proliferation under hypoxia. Furthermore, both HIF-1a and N-Myc are essential for theWarburg effect (aerobic glycolysis) in neuroblastomas by activating the transcription of multiple glycolytic genes. Of note, expressions of Phosphoglycerate Kinase 1 (PGK1), Hexokinase 2 (HK2), and Lactate Dehydrogenase A (LDHA) were each significantly higher in MYCNamplified neuroblastomas than in tumors without MYCN amplification. Interestingly, MYCN-amplified neuroblastoma cells are "addicted" to LDHA enzymatic activity, as its depletion completely inhibits tumorigenesis in vivo. Thus, our results providemechanistic insights explaining howMYCN-amplified neuroblastoma cells contend with hypoxic stress and paradoxically how hypoxia contributes to neuroblastoma aggressiveness through combinatorial effects of N-Myc and HIF-1a. These results also suggest that LDHA represents a novel, pharmacologically tractable target for neuroblastoma therapeutics. Cancer Res; 70(24); 10351–61. 2010 AACR.