Emerging Utility of Urinary Cell-free Nucleic Acid Biomarkers for Prostate, Bladder, and Renal Cancers

Lin, Selena Y.; Linehan, Jennifer; Wilson, Timothy G.; Hoon, Dave S.B.

doi:10.1016/j.euf.2017.03.009

Cited by 62 publications

(50 citation statements)

References 48 publications

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“…Urine is a complex liquid containing EVs, protein, nucleic acids and many other metabolites (30). Tamm-Horsfell is a protein in urine that is known to form networks that trap EVs (31).…”

Section: Methodsmentioning

confidence: 99%

A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis

Zhou

Spillman

Behbakht

et al. 2017

Analytical Biochemistry

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Readily accessible samples such as urine or blood are seemingly ideal for differentiating and stratifying patients; however, it has proven a daunting task to identify reliable biomarkers in such samples. Noncoding RNA holds great promise as a source of biomarkers distinguishing physiologic wellbeing or illness. Current methods to isolate and characterize RNA molecules in urine are limited. In this proof of concept study, we present a method to extract and identify small noncoding RNAs in urine. Initially, quantitative reverse transcription PCR was applied to confirm the presence of microRNAs in total RNA extracted from urine. Once the presence of micro RNA in urine was confirmed, we developed a method to scale up RNA extraction to provide adequate amounts of RNA for next generation sequence analysis. The method described in this study is applicable to detecting a broad range of small noncoding RNAs in urine; thus, they have wide applicability for health and disease analyses.

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“…Urine is a complex liquid containing EVs, protein, nucleic acids and many other metabolites (30). Tamm-Horsfell is a protein in urine that is known to form networks that trap EVs (31).…”

Section: Methodsmentioning

confidence: 99%

A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis

Zhou

Spillman

Behbakht

et al. 2017

Analytical Biochemistry

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“…To compare different sample storage conditions for minimizing cfDNA loss, urine supernatant from 10 healthy individuals was aliquoted into four tubes and kept under four different conditions: (1) storage at −20 • C with the addition of 10 mM EDTA, (2) storage at −20 • C without the addition of EDTA, (3) storage at −70 • C with the addition of 10 mM EDTA, and (4) storage at −70 • C without the addition of EDTA ( Figure 3). EDTA was used as the preservative as it could be easily used to inactivate nuclease activity [13]. After three months, the frozen sample was thawed at 4 • C and centrifuged at 3000 g for 10 min and was then subjected to cfDNA isolation.…”

Section: Comparison Of Sample Storage Conditionsmentioning

confidence: 99%

“…DNA fragments in the region of over 1000 bp were considered to be derived from urinary cellular DNA [13], and the DNA concentration of high molecular weight (HMW) DNA above 1 kb was measured to examine the degree of contamination by cellular DNA. Figure 3B shows the yield ratio of 50-300 bp cfDNA to HMW DNA obtained from 1 mL of urine.…”

Section: Contamination Of Cellular Genomic Dnamentioning

confidence: 99%

“…Specifically, urine could be more useful in liquid biopsy for urologic malignant disease. Several studies have demonstrated that urinary cfDNA can be utilized as an important source for liquid biopsy in urologic malignant disease [5,[11][12][13][14]. Due to the increased rate of necrosis of tumor cells, urinary cfDNA carries a higher tumor genomic burden than cellular DNA [11,15].…”

Section: Introductionmentioning

confidence: 99%

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Comparison of Four Commercial Kits for Isolation of Urinary Cell-Free DNA and Sample Storage Conditions

Lee

Yoon

et al. 2020

Diagnostics

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Urinary cell-free DNA (cfDNA) is an attractive body fluid for liquid biopsy. In this study, we compared the efficiencies of four commercial kits for urinary cell-free DNA (cfDNA) isolation and of various sample storage conditions. Urinary cfDNA was isolated from 10 healthy individuals using four commercial kits: QIAamp Circulating Nucleic Acid Kit (QC; Qiagen), MagMAX™ Cell-Free DNA Isolation Kit (MM; Applied Biosystems), Urine Cell-Free Circulating DNA Purification Midi Kit (NU; Norgen Biotek), and Quick-DNA™ Urine Kit (ZQ; Zymo Research). To assess the isolation efficiency, an Agilent 2100 Bioanalyzer with High Sensitivity DNA chips was used, and cfDNA yield was defined as the amount of cfDNA obtained from 1 mL of urine. MM and QC provided the highest cfDNA yield in the 50–300 bp range, and MM and NU gave the highest cfDNA yield in the 50–100 bp range. In particular, the NU kit was efficient for isolation of more fragmented cfDNA in the range of 50–100 bp with the lowest cellular genomic DNA contamination. ZQ had the best cost-efficiency for isolating the same amount of urinary cfDNA. Samples stored at −70 °C with the addition of 10 mM EDTA resulted in the highest cfDNA yield 3 months after sample collection.

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“…The scarce information available, although to be further corroborated, would suggest the potential employment of these proteins as biomarkers of these neoplastic diseases, whose incidence continues to rapidly increase worldwide and for the early detection and follow up of which no biomarkers are currently available [14]. RCa and BCa lack specific predictive biomarkers and only some symptoms might have some effects in finding the existence of cancer [72,73]. The study of the role of glyoxalases in TCa is today pratically still a "virgin field".…”

Section: Concluding Remarks and Future Directionsmentioning

confidence: 99%

Glyoxalases in Urological Malignancies

Antognelli¹,

Talesa²

2017

Preprint

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Urological cancers include a spectrum of malignancies affecting organs of the reproductive and/or urinary systems, such as prostate, kidney, bladder, and testis. Despite improved primary prevention, detection and treatment, urological cancers are still characterized by an increasing incidence and mortality worldwide and although advances have been made toward understanding the molecular basis of these diseases, a complete insight of the pathogenic mechanisms is still a research challenge for defining safer pharmacological therapies and prognostic factors, especially for the metastatic stage of these neoplasms for which no effective therapies exist. Glyoxalases are enzymes that catalyzes the glutathione-dependent metabolism of cytotoxic methylglyoxal (MG), thus protecting against cellular damage and apoptosis. They are generally overexpressed in numerous cancers as a survival strategy by providing a safeguard through enhancement of MG detoxification. Increasing evidence suggest that glyoxalases, especially Glo1, would act as oncogenes in urological malignancies and be central to their initiation, growth and progression. In this review, we highlight the critical role of glyoxalases as regulators of tumorigenesis in the prostate through modulation of various critical signaling pathways, and provide an overview of the current knowledge on glyoxalases in bladder, kidney and testis cancers. We also discuss the promise and challenges for Glo1 inhibitors as future anti-PCa therapeutics and the potential of glyoxalases as biomarkers for PCa diagnosis.

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Emerging Utility of Urinary Cell-free Nucleic Acid Biomarkers for Prostate, Bladder, and Renal Cancers

Cited by 62 publications

References 48 publications

A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis

A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis

Comparison of Four Commercial Kits for Isolation of Urinary Cell-Free DNA and Sample Storage Conditions

Glyoxalases in Urological Malignancies

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