Ultrasensitive and Reversible Nanoplatform of Urinary Exosomes for Prostate Cancer Diagnosis

Abstract

Prostate cancer cell-derived exosomes in urine have been extensively studied recently and regarded as novel biomarkers for cancer diagnosis and prognosis, which presents wide prospects in clinical applications. Sensitive detection and specific capture methods are essential for exosomes analysis. Herein, a dual functional platform composed of superparamagnetic conjunctions and molecular beacons (SMC-MB) is reported. The SMC-MB platform is designed based on aptamer immunoaffinity with ultrasensitive detection efficiency and reversible isolation capacity, which, respectively, profit from nonenzymatic amplification methods and magnetic separation along with restriction cleavage. It is noteworthy that exosomes quantification was exactly amplified and transformed into single strand DNA detection. Correlated measurements evidence that the limit of detection of SMC-MB is as low as ∼100 particles/μL in urine, and a linear relationship meets between the logarithmic concentration of exosomes and fluorescence intensity of the molecular beacon. Furthermore, employing prostate specific membrane antigen (PSMA) aptamer, the platform adapted to detect and capture PMSA-positive exosomes from urine samples provides excellent diagnostic efficiency for prostate cancer (PCa). The expression of typical biomarkers of PCa, i.e., PSA and PCA3 mRNA, is significantly higher in PSMA-positive exosomes. Altogether, the platform and strategy described in this paper are promising in urinary exosomes analysis and prostate cancer detection.

Isolation and Detection Technologies of Extracellular Vesicles and Application on Cancer Diagnostic

“…Reprinted with permission from Yang et al 130 Copyright © 1999-2019 John Wiley & Sons, Inc. B, Histogram and boxplot of fluorescence intensity of exosomes (Target: Prostate-specific membrane antigen (PSMA) positive) from patients with prostate cancer and healthy donors detected by superparamagnetic conjunctions and molecular beacons (SMC-MB) platform. Reprinted with permission from Li et al 134 Copyright © 2019 American Chemical Society. C, Comparison of glypican-1 expression level in patients with pancreatic cancer (n = 20), benign pancreatic disease (n = 7), and healthy controls (n = 11).…”

“…146 Li et al used an ultrasensitive and reversible nanoplatform to detect PSA, PCA3, and mRNA successfully in urinary exosomes from patients with PCa ( Figure 2B). 134 However, PSA has limitation because it may not differ cancer and benign prostatic hyperplasia (BPH). 146 Some miRNAs, such as miR-141 and miR-375, from serum EVs were associated with metastatic PCa, and the miR-19b distinguished PCa with 100% specificity and 93% sensitivity.…”

Integrin Alpha V in Urine: A Novel Noninvasive Marker for Prostate Cancer Detection

“…To prepare conditioned medium (CM), 1×10 6 LnCaP cells were first plated in complete medium and grown for 24 h. Cells were then washed twice with PBS and placed in a serum-free medium. CM was collected after 48 h, centrifuged at 3,000 rpm to pellet debris, and filtered through a 0.1-micron filter using ultrafiltration toolkit Exo:pure (Prostagnost, Russian Federation) to separate the medium from microvesicles.…”

“…The single urinary PCA3 test has been approved so far for clinical use. Studies of microRNAs (miRNAs) and exosomes isolated from urine is another area to search for biomarkers, which could be promising in pre-biopsy prediction of PCa (3)(4)(5)(6). Recently, integrated analysis of metabolomic and transcriptomic data obtained from urine samples of PCa patients, BPH, and healthy individuals revealed abnormal glutamate metabolism and tricarboxylic acid cycle in the prostate cancer (7).…”

Liquid biopsies in myeloid malignancies

“…As exosomes content depends on the physiological state of the cell, the cargo and abundance of tumor derived exosomes generally reflects the stage of the tumor, rendering diagnostic value in LB for early tumor detection [153][154][155] . Even though exosomes are not being used in clinical practice, the potential of these vesicles for diagnosis is highlighted by the 71 clinical trials studies focused on exosomes of cancer patients (according to www.clinicalTrials.gov accessed on 12 September 2019), accompanied by the development of platforms for tumor derived exosomes detection in complex samples (e.g., surface plasmon resonance for exosome biomarkers detection [156] , microfluidic chip for ovarian cancer diagnosis [157] , or a platform with superparamagnetic conjunctions and molecular beacons targeting urinary exosomes for prostate cancer diagnosis [158] ). Nevertheless, several sensitive and specific exosomal biomarkers were already suggested for early diagnosis of ovarian cancer (over-expression of miR-200a, miR-200b, and miR200c in patient's serum exosomes) [159][160][161] , non-small cell lung cancer (enrichment of Leucine-rich a2-glycoprotein 1 in exosomes of patients urine and MALAT-1 in patient's blood) [162][163][164] , colon cancer (overexpression of miR-1246 and miR-23a in patient's serum exosomes) [165,166] , or pancreatic ductal adenocarcinoma (presence of glypican-1 in patient's serum) [167,168] .…”