Rapid and Label-Free Isolation of Tumour Cells from the Urine of Patients with Localised Prostate Cancer Using Inertial Microfluidics

Abstract

During the last decade, isolation of circulating tumour cells via blood liquid biopsy of prostate cancer (PCa) has attracted significant attention as an alternative, or substitute, to conventional diagnostic tests. However, it was previously determined that localised forms of PCa shed a small number of cancer cells into the bloodstream, and a large volume of blood is required just for a single test, which is impractical. To address this issue, urine has been used as an alternative to blood for liquid biopsy as a truly non-invasive, patient-friendly test. To this end, we developed a spiral microfluidic chip capable of isolating PCa cells from the urine of PCa patients. Potential clinical utility of the chip was demonstrated using anti-Glypican-1 (GPC-1) antibody as a model of the primary antibody in immunofluorescent assay for identification and detection of the collected tumour cells. The microchannel device was first evaluated using DU-145 cells in a diluted Dulbecco’s phosphate-buffered saline sample, where it demonstrated >85 (±6) % efficiency. The microchannel proved to be functional in at least 79% of cases for capturing GPC1+ putative tumour cells from the urine of patients with localised PCa. More importantly, a correlation was found between the amount of the captured GPC1+ cells and crucial diagnostic and prognostic parameter of localised PCa—Gleason score. Thus, the technique demonstrated promise for further assessment of its diagnostic value in PCa detection, diagnosis, and prognosis.

The Glycocalyx and Its Role in Vascular Physiology and Vascular Related Diseases

“…Mediates cell adhesion to extracellular matrix 52 , 86 Glypicans Glypican-1 Upregulated in lymphoma, breast, pancreatic, and cervical cancer. Modulates mitogenic response 19 , 46 , 54 , 72 , 97 , 110 , 111 , 117 , 126 , 127 , 131 , 166 , 177 , 178 , 218 Glypican-3 Upregulated in hepatocellular, gastric, ovarian and lung squamous cell carcinoma; downregulated in breast, lung, ovary, and mesothelium cancers 67 , 86 , 153 …”

“… 178 The use of GPC1 antibodies as a diagnostic tool to detect circulating tumor cells and exosomes, as well as labeling of cancer cells in situ , is an active area of investigation. 19 , 110 , 131 , 166 , 177 , 218 …”

Urine biopsy technologies: Cancer and beyond

“…Studies have found that the cfDNA released by cells into the systemic circulation can also be filtered by the kidneys and transmitted in urine 7 , 8 . Therefore, scientists and clinicians are keen to utilize urine biopsies for the potential detection of diseases, including colorectal cancer (CC) 3 , bladder cancer (BC) 9 , chronic kidney disease 10 , prostate cancer (PC) 11 , cystic fibrosis 12 , and chronic obstructive pulmonary disease 13 .…”

Liquid Biopsy Biomarkers in Urine: A Route towards Molecular Diagnosis and Personalized Medicine of Bladder Cancer

“…Several types of cells are present in urine, namely epithelial cells, renal-derived cells, leukocytes, erythrocytes and urothelial cells, as well as genetic material, proteins, peptides and inorganic elements. Therefore, scientific and medical personnel prefer to use urinary biopsies for the potential diagnosis of diseases, such as colorectal cancer (CC) [ 4 ], bladder cancer [ 3 ], chronic renal disease [ 6 ], prostate cancer [ 7 ], cystic fibrosis [ 8 ] and chronic obstructive pulmonary disease [ 9 ]. Urine may be used in its entirety or divided into supernatant and pellet for biomarker analysis.…”