Abstract
Background: Liquid biopsies offer a promising alternative to tissue samples, providing non-invasive diagnostic approaches or serial monitoring of disease evolution. However, certain challenges remain, and the full potential of liquid biopsies has yet to be reached. Here we report several methodological approaches to interrogate liquid biopsies using circulating tumour cell (CTC) enumeration and characterisation, transcriptomics, Raman spectroscopy, and copy number instability (CNI) scores using blood samples of lung cancer (LC) patients. Methods: We choose LC; since it still is the most common cause of cancer-related mortality worldwide, and therefore there is a need for development of new non-invasive diagnostic/prognostic technologies. Changes in gene expression were assessed using RNA-seq, and in CTCs using ImageStream, an imaging flow-cytometer. CNI scores, from paired tissue/ctDNA were also explored. Raman spectroscopy was used to provide chemical fingerprints of plasma samples. Results: CTCs were detected in all LC patients (n = 10). We observed a significant increase in CTC levels in LC patients (n = 10) compared to controls (n = 21). A similar CNI was noted in the tissue and plasma of 2 patients, where higher CNI scores corresponded with poorer outcome. Significant changes in Raman spectra (carotenoid concentrations) were noted in LC patients (n = 20) compared to controls (n = 10). RNA-seq revealed differential expression of 21 genes between LC cases and controls in both LC tissue and blood samples. Conclusions: Liquid biopsies can potentially provide a more comprehensive picture of the disease compared to a single tissue biopsy. CTC enumeration is feasible and sensitive for LC patients. Molecular profiling of CTCs is also possible from total blood. CNI scores and Raman spectra require further investigation. Further work is being undertaken to explore these methods of detection in a larger LC cohort.
Highlights
Over the past decade, the concept of liquid biopsies has been introduced as an alternative to a conventional needle biopsies for several cancers
We have demonstrated a potential use of Alu repeats ratios for prognostic purposes in the advanced setting for lung cancer (LC) patients [14]
We have demonstrated that measuring the chemical composition plasma using Raman spectroscopy can be of diagnostic value
Summary
The concept of liquid biopsies has been introduced as an alternative to a conventional needle (tissue) biopsies for several cancers. Tests can be conducted on a blood sample (i.e., non-invasively) to examine circulating tumour cells (CTCs) or circulating tumour DNA (ctDNA) with the aim to provide a better insight into the real-time dynamics of a disease. CTCs are tumour cells present in the bloodstream that shed either from the primary tumour or its metastasis [3,4] These cells are different to normal circulating blood cells and express tumour-specific characteristics [5,6]. Liquid biopsies offer a promising alternative to tissue samples, providing non-invasive diagnostic approaches or serial monitoring of disease evolution. Enumeration and characterisation, transcriptomics, Raman spectroscopy, and copy number instability (CNI) scores using blood samples of lung cancer (LC) patients.
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