Aldehyde dehydrogenase activity plays a Key role in the aggressive phenotype of neuroblastoma

Abstract

BackgroundThe successful targeting of neuroblastoma (NB) by associating tumor-initiating cells (TICs) is a major challenge in the development of new therapeutic strategies. The subfamily of aldehyde dehydrogenases 1 (ALDH1) isoenzymes, which comprises ALDH1A1, ALDH1A2, and ALDH1A3, is involved in the synthesis of retinoic acid, and has been identified as functional stem cell markers in diverse cancers. By combining serial neurosphere passages with gene expression profiling, we have previously identified ALDH1A2 and ALDH1A3 as potential NB TICs markers in patient-derived xenograft tumors. In this study, we explored the involvement of ALDH1 isoenzymes and the related ALDH activity in NB aggressive properties.MethodsALDH activity and ALDH1A1/A2/A3 expression levels were measured using the ALDEFLUOR™ kit, and by real-time PCR, respectively. ALDH activity was inhibited using the specific ALDH inhibitor diethylaminobenzaldehyde (DEAB), and ALDH1A3 gene knock-out was generated through the CRISPR/Cas9 technology.ResultsWe first confirmed the enrichment of ALDH1A2 and ALDH1A3 mRNA expression in NB cell lines and patient-derived xenograft tumors during neurosphere passages. We found that high ALDH1A1 expression was associated with less aggressive NB tumors and cell lines, and correlated with favorable prognostic factors. In contrast, we observed that ALDH1A3 was more widely expressed in NB cell lines and was associated with poor survival and high-risk prognostic factors. We also identified an important ALDH activity in various NB cell lines and patient-derived xenograft tumors. Specific inhibition of ALDH activity with diethylaminobenzaldehyde (DEAB) resulted in a strong reduction of NB cell clonogenicity, and TIC self-renewal potential, and partially enhanced NB cells sensitivity to 4-hydroxycyclophosphamide. Finally, the specific knock-out of ALDH1A3 via CRISPR/Cas9 gene editing reduced NB cell clonogenicity, and mediated a cell type-dependent inhibition of TIC self-renewal properties.ConclusionsTogether our data uncover the participation of ALDH enzymatic activity in the aggressive properties and 4-hydroxycyclophosphamide resistance of NB, and show that the specific ALDH1A3 isoenzyme increases the aggressive capacities of a subset of NB cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2820-1) contains supplementary material, which is available to authorized users.

Imperative role of particulate matter in innate immunity during RNA virus infection

“…Additionally, induction of CYP1A1 in presence of diesel exhaust particles were extensively reported in human bronchial cells [58]. Apart from studies related to different types of cancers [59, 60], ALDH1A3 was also previously reported in connection with virus infections like human papilloma virus and respiratory syncytial virus [61-63]. Altogether, these significant differentially expressed genes noted in our study related to different metabolic modifications inside the cell and reasonably linked to virus infections, therefore, we selected these genes for validation in context to RNA virus infectivity.…”

Neuroblastoma pathogenesis: deregulation of embryonic neural crest development

“…shRNA knockdown of ALDH1A2 reduced colony and tumorsphere formation in vitro (Hartomo, et al, 2015). Similarly, inhibition and knockout of isoform ALDH1A3 led to reduction in NB tumor initiating cells (TICs) as demonstrated by reduced colony formation and reduced tumorsphere passaging (Flahaut, et al, 2016). However, despite these in vitro results, when ALDH12A knockdown cell lines were injected subcutaneously into mice, there was no difference in tumor formation and only a slight, non-significant difference in tumor size (Hartomo, et al, 2015).…”

Neuroblastoma: Genetic, Epigenetic Mechanisms and Targeted Therapy Approaches: Review

“…ALDH1 izoenzimlerinin aktivitesi, CRISPR/ Cas9 teknolojisi kullanılarak hastadan köken alan ksenograft tümörleri kullanan çalışmada, nöroblastom kök hücrelerinin agresif doğasıyla ilişkilendirilmiştir. 96 Hayvan modellerinde, CRISPR/Cas9 teknolojisi kullanan DNMT3a transaktivasyonu, beyin hücrelerinin metilasyonunun düzenlenmesine katkıda bulunmaktadır. 97 CRISPR/Cas9 teknolojisini kullanarak NRAS mutant hücre hattında rehber RNA (gRNA) lar aracılığıyla mutant NRAS geninin susturulması, hücreleri spesifik inhibitörlere karşı daha duyarlı hâle getirmiştir.…”