Development and validation of a 25-Gene Panel urine test for prostate cancer diagnosis and potential treatment follow-up

Abstract

Background Heterogeneity of prostate cancer (PCa) contributes to inaccurate cancer screening and diagnosis, unnecessary biopsies, and overtreatment. We intended to develop non-invasive urine tests for accurate PCa diagnosis to avoid unnecessary biopsies. Methods Using a machine learning program, we identified a 25-Gene Panel classifier for distinguishing PCa and benign prostate. A non-invasive test using pre-biopsy urine samples collected without digital rectal examination (DRE) was used to measure gene expression of the panel using cDNA preamplification followed by real-time qRT-PCR. The 25-Gene Panel urine test was validated in independent multi-center retrospective and prospective studies. The diagnostic performance of the test was assessed against the pathological diagnosis from biopsy by discriminant analysis. Uni- and multivariate logistic regression analysis was performed to assess its diagnostic improvement over PSA and risk factors. In addition, the 25-Gene Panel urine test was used to identify clinically significant PCa. Furthermore, the 25-Gene Panel urine test was assessed in a subset of patients to examine if cancer was detected after prostatectomy. Results The 25-Gene Panel urine test accurately detected cancer and benign prostate with AUC of 0.946 (95% CI 0.963–0.929) in the retrospective cohort (n = 614), AUC of 0.901 (0.929–0.873) in the prospective cohort (n = 396), and AUC of 0.936 (0.956–0.916) in the large combination cohort (n = 1010). It greatly improved diagnostic accuracy over PSA and risk factors (p < 0.0001). When it was combined with PSA, the AUC increased to 0.961 (0.980–0.942). Importantly, the 25-Gene Panel urine test was able to accurately identify clinically significant and insignificant PCa with AUC of 0.928 (95% CI 0.947–0.909) in the combination cohort (n = 727). In addition, it was able to show the absence of cancer after prostatectomy with high accuracy. Conclusions The 25-Gene Panel urine test is the first highly accurate and non-invasive liquid biopsy method without DRE for PCa diagnosis. In clinical practice, it may be used for identifying patients in need of biopsy for cancer diagnosis and patients with clinically significant cancer for immediate treatment, and potentially assisting cancer treatment follow-up.

Myosin 1C isoform A is a novel candidate diagnostic marker for prostate cancer

“…Validation of biomarkers suitable for non-invasive diagnostics using patients' serum and urine is an important direction in the current research on PCa [27][28][29]. In the light of the present findings that establish utility of myosin IC isoform A in analyzing biopsy samples and our recent results [11] that demonstrated presence of myosin IC isoform A in exosomes secreted…”

“…Validation of biomarkers suitable for non-invasive diagnostics using patients’ serum and urine is an important direction in the current research on PCa [ 27 – 29 ]. In the light of the present findings that establish utility of myosin IC isoform A in analyzing biopsy samples and our recent results [ 11 ] that demonstrated presence of myosin IC isoform A in exosomes secreted by PCa cells, assessing diagnostic applications of this isoform’s detection in serum or urine is a logical next step in validation of this biomarker for clinical use.…”

“…Expression of myosin IC isoform A, according to the data presented here, is a robust and differentiating feature of clinical samples from the different stages of the disease, and thus compares favorably with markers that are detected in small subgroups of patients (e.g., the recently characterized isoform of NCOR1, [ 30 ]). Assembly of biomarker panels with high collective diagnostic capacity is another direction of current work in the field [ 29 ]. The ability to differentiate stages of PCa progression that is demonstrated here raises the likelihood of eventually including myosin IC isoform A in a panel of biomarkers that will be capable of accurately and predictively diagnosing PCa and the likely course of the disease.…”

A 23‐Gene Classifier urine test for prostate cancer prognosis

“…Xuhui Zhang 4 Xiaoyan Feng 4 Heqiu Zhang 4 Athanasios Simoulis 5 Alan HB Wu 6 Taolin Xia 7 Fei Li 8 Wanlong Tan 8 Allan Johnson 9 Nishtman Dizeyi 10 Per-Anders Abrahamsson 10 Lukas Kenner 11 Lingwu Chen 12 Wanmei Zhong 12 Kefeng Xiao 1,2 Jenny L. Persson 13,14,15 Chang Zou…”

Non-invasive Urine Test for Molecular Classification of Clinical Significance in Newly Diagnosed Prostate Cancer Patients

“…The procedures for urine sample processing and gene expression quantification were performed as described (20). In the retrospective study, 10-15 ml urine samples were collected without prior digital rectal examination and the urine pellet was flash-frozen and stored at −80 • C. In the prospective study, 15-45 ml urine was collected without prior DRE and the urine was stored with 5 ml DNA/RNA preservative AssayAssure (Thermo Fisher Scientific, Waltham, MA, USA) or U-Preserve (Hao Rui Jia Biotech Ltd., Beijing, China) at 4 • C and processed within a week.…”