Nanopore Sequencing Reveals Global Transcriptome Signatures of Mitochondrial and Ribosomal Gene Expressions in Various Human Cancer Stem-like Cell Populations

SIMPLE SUMMARY: Cancer is the leading cause of death in the industrialized world. In particular, so-called cancer stem cells (CSCs) play a crucial role in disease progression, as they are known to contribute to tumor growth and metastasis. Thus, CSCs are heavily investigated in a broad range of cancers. Nevertheless, global transcriptomic profiling of CSC populations derived from different tumor types is rare. We established three CSC populations from tumors in the uterus, brain, lung, and prostate and assessed their global transcriptomes using nanopore full-length cDNA sequencing, a new technique to assess insights into global gene profile. We observed common expression in all CSCs for distinct genes encoding proteins for organelles, such as ribosomes, mitochondria, and proteasomes. Additionally, we detected high expressions of inflammation- and immunity-related genes. Conclusively, we observed high similarities between all CSCs independent of their tumor of origin, which may build the basis for identifying novel therapeutic strategies targeting CSCs. ABSTRACT: Cancer stem cells (CSCs) are crucial mediators of tumor growth, metastasis, therapy resistance, and recurrence in a broad variety of human cancers. Although their biology is increasingly investigated within the distinct types of cancer, direct comparisons of CSCs from different tumor types allowing comprehensive mechanistic insights are rarely assessed. In the present study, we isolated CSCs from endometrioid carcinomas, glioblastoma multiforme as well as adenocarcinomas of lung and prostate and assessed their global transcriptomes using full-length cDNA nanopore sequencing. Despite the expression of common CSC markers, principal component analysis showed a distinct separation of the CSC populations into three clusters independent of the specific type of tumor. However, GO-term and KEGG pathway enrichment analysis revealed upregulated genes related to ribosomal biosynthesis, the mitochondrion, oxidative phosphorylation, and glycolytic pathways, as well as the proteasome, suggesting a great extent of metabolic flexibility in CSCs. Interestingly, the GO term “NF-kB binding” was likewise found to be elevated in all investigated CSC populations. In summary, we here provide evidence for high global transcriptional similarities between CSCs from various tumors, which particularly share upregulated gene expression associated with mitochondrial and ribosomal activity. Our findings may build the basis for identifying novel therapeutic strategies targeting CSCs.