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- Title
Plasma Next Generation Sequencing and Droplet Digital-qPCR-Based Quantification of Circulating Cell-Free RNA for Noninvasive Early Detection of Cancer.
- Authors
Metzenmacher, Martin; Váraljai, Renáta; Hegedüs, Balazs; Cima, Igor; Forster, Jan; Schramm, Alexander; Scheffler, Björn; Horn, Peter A.; Klein, Christoph A.; Szarvas, Tibor; Reis, Hennig; Bielefeld, Nicola; Roesch, Alexander; Aigner, Clemens; Kunzmann, Volker; Wiesweg, Marcel; Siveke, Jens T.; Schuler, Martin; Lueong, Smiths S.
- Abstract
Early detection of cancer holds high promise for reducing cancer-related mortality. Detection of circulating tumor-specific nucleic acids holds promise, but sensitivity and specificity issues remain with current technology. We studied cell-free RNA (cfRNA) in patients with non-small cell lung cancer (NSCLC; n = 56 stage IV, n = 39 stages I-III), pancreatic cancer (PDAC, n = 20 stage III), malignant melanoma (MM, n = 12 stage III-IV), urothelial bladder cancer (UBC, n = 22 stage II and IV), and 65 healthy controls by means of next generation sequencing (NGS) and real-time droplet digital PCR (RT-ddPCR). We identified 192 overlapping upregulated transcripts in NSCLC and PDAC by NGS, more than 90% of which were noncoding. Previously reported transcripts (e.g., HOTAIRM1) were identified. Plasma cfRNA transcript levels of POU6F2-AS2 discriminated NSCLC from healthy donors (AUC = 0.82 and 0.76 for stages IV and I–III, respectively) and significantly associated (p = 0.017) with the established tumor marker Cyfra 21-1. cfRNA yield and POU6F2-AS transcript abundance discriminated PDAC patients from healthy donors (AUC = 1.0). POU6F2-AS2 transcript was significantly higher in MM (p = 0.044). In summary, our findings support further validation of cfRNA detection by RT-ddPCR as a biomarker for early detection of solid cancers.
- Subjects
LUNG cancer treatment; ANTINEOPLASTIC agents; BIOMARKERS; BIOCHEMISTRY; BLADDER tumors; CLINICAL drug trials; PHENOMENOLOGY; PANCREATIC tumors; POLYMERASE chain reaction; RNA; TUMOR classification; DESCRIPTIVE statistics; SEQUENCE analysis; EARLY detection of cancer
- Publication
Cancers, 2020, Vol 12, Issue 2, p353
- ISSN
2072-6694
- Publication type
Article
- DOI
10.3390/cancers12020353