Comprehensive Approach to Distinguish Patients with Solid Tumors from Healthy Controls by Combining Androgen Receptor Mutation p.H875Y with Cell-Free DNA Methylation and Circulating miRNAs.

Clemens Heitzinger,Olivier J Switzeny,Elena Tomeva,Alexander G Haslberger,Berit Hippe

doi:10.3390/cancers14020462

Clemens Heitzinger, Olivier J Switzeny + Show 3 more

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https://doi.org/10.3390/cancers14020462

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Abstract

Simple SummaryBlood-based tests for cancer detection are minimally invasive and could be useful for screening asymptomatic patients and high-risk populations. Since a single molecular biomarker is usually insufficient for an accurate diagnosis, we developed a multi-analyte liquid biopsy-based classification model to distinguish cancer patients from healthy subjects. The combination of cell-free DNA mutations, miRNAs, and cell-free DNA methylation markers improved the model’s performance. Moreover, we demonstrated that the androgen receptor mutation p.H875Y is not only relevant in prostate cancer but had a strong predictive value for colorectal, bladder, and breast cancer. Our results, although preliminary, showed that a single liquid biopsy test could detect multiple cancer types simultaneously.Liquid biopsy-based tests emerge progressively as an important tool for cancer diagnostics and management. Currently, researchers focus on a single biomarker type and one tumor entity. This study aimed to create a multi-analyte liquid biopsy test for the simultaneous detection of several solid cancers. For this purpose, we analyzed cell-free DNA (cfDNA) mutations and methylation, as well as circulating miRNAs (miRNAs) in plasma samples from 97 patients with cancer (20 bladder, 9 brain, 30 breast, 28 colorectal, 29 lung, 19 ovarian, 12 pancreas, 27 prostate, 23 stomach) and 15 healthy controls via real-time qPCR. Androgen receptor p.H875Y mutation (AR) was detected for the first time in bladder, lung, stomach, ovarian, brain, and pancreas cancer, all together in 51.3% of all cancer samples and in none of the healthy controls. A discriminant function model, comprising cfDNA mutations (COSM10758, COSM18561), cfDNA methylation markers (MLH1, MDR1, GATA5, SFN) and miRNAs (miR-17-5p, miR-20a-5p, miR-21-5p, miR-26a-5p, miR-27a-3p, miR-29c-3p, miR-92a-3p, miR-101-3p, miR-133a-3p, miR-148b-3p, miR-155-5p, miR-195-5p) could further classify healthy and tumor samples with 95.4% accuracy, 97.9% sensitivity, 80% specificity. This multi-analyte liquid biopsy-based test may help improve the simultaneous detection of several cancer types and underlines the importance of combining genetic and epigenetic biomarkers.

Highlights

Cancer is mostly a manageable disease as long as it is diagnosed and treated before metastasis has begun
One of the cancer samples was excluded since the patient was diagnosed with stage 4 ovarian cancer after plasma collection
Our results suggest that plasma Cell-free DNA (cfDNA) could serve rather as a monitor for disease progression since cfDNA concentration correlated with cancer stage regardless of cancer type

Summary

Introduction

Cancer is mostly a manageable disease as long as it is diagnosed and treated before metastasis has begun. Early tumor detection could increase the chances of successful treatment. In this way, carcinomas could be identified at an early stage when they can still be surgically removed and cured [1,2]. Liquid biopsy is a rapidly developing tool for assessing biomarkers shed from difficult to access tissue in sampled bodily fluids, such as urine, blood, saliva, sweat, feces, and tears [3,4]. Such minimally invasive blood-based tests could be useful for screening asymptomatic patients and high-risk populations. Liquid biopsy-based tests have been successfully applied in disease screening [5] but tend to have even higher compliance in comparison to other standard procedures such as FIT (fecal immunochemical test) [6]

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