Editorial

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Open AccessEditorial

Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis

by Paola Ulivi and Stefano Indraccolo

Biomedicines 2022, 10(11), 2748; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines10112748 - 29 Oct 2022

Cited by 2 | Viewed by 1121

Liquid biopsy has emerged as new tool for detecting clinically relevant genetic alterations in cancer patients [...] Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

Research

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14 pages, 23182 KiB

Open AccessArticle

Cancer Detection Using an Artificial Secretable MicroRNA Found in Blood and Urine

by Pei-Wei Shueng, Kuang-Chung Shih, Sanjiv Sam Gambhir, Deng-Yu Kuo and Hui-Yen Chuang

Biomedicines 2022, 10(3), 621; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines10030621 - 07 Mar 2022

Cited by 2 | Viewed by 2180

Abstract

Biomarkers can potentially help in the detection and prognosis of diseases such as cancer, its recurrence, predicting response to therapy, and monitoring of response during and/or after treatment. Endogenous tumor blood biomarkers suffer from low concentrations that are not distinguishable from background noise [...] Read more.

Biomarkers can potentially help in the detection and prognosis of diseases such as cancer, its recurrence, predicting response to therapy, and monitoring of response during and/or after treatment. Endogenous tumor blood biomarkers suffer from low concentrations that are not distinguishable from background noise and, if identified, the localization of the biomarker production site is not known. The use of exogenously introduced or artificial biomarkers can eliminate these issues. In this study, we show that cancer cells can be made to produce an artificial secreted microRNA (Sec-miR) that can be detected in media from cells in culture, and from both blood and urine in living mice. In culture, we show that chaining a number of Sec-miR sequences in a plasmid and transfecting cells with the plasmids could increase Sec-miR secretion as the number of sequences increases. Tumor induction in mice with a stably transfected HeLa cell line shows the presence and significant increase in the Sec-miR with time and tumor growth in plasma (p < 0.001, R2 = 0.5542). The relative half-life of the Sec-miR was seen to be 1.2 h in the plasma of living mice and was seen to appear in urine within 12 h. The transgene for the Sec-miR within a minicircle was introduced via the tail-vein into subcutaneous tumor-bearing mice. As the tumor growth increased with time, further in vivo transfection of the Sec-miR minicircles showed an increase in Sec-miR in both plasma and urine (R2 = 0.4546). This study demonstrated that an exogenous Sec-miR biomarker would allow for early tumor detection using in vitro diagnostics techniques. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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8 pages, 761 KiB

Open AccessArticle

Pretherapeutic Serum Albumin as an Outcome Prognosticator in Head and Neck Adenoid-Cystic Carcinoma

by Marlene Friedl, Stefan Stoiber, Faris F. Brkic and Lorenz Kadletz-Wanke

Biomedicines 2022, 10(1), 191; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines10010191 - 17 Jan 2022

Cited by 4 | Viewed by 1739

Abstract

Background: A head and neck adenoid-cystic carcinoma is a rare malignant tumor arising from the salivary gland tissues. The long-term survival outcome is poor due to a high risk of recurrences and distant metastasis. The identification of prognostic markers could contribute to a [...] Read more.

Background: A head and neck adenoid-cystic carcinoma is a rare malignant tumor arising from the salivary gland tissues. The long-term survival outcome is poor due to a high risk of recurrences and distant metastasis. The identification of prognostic markers could contribute to a better risk assessment of each patient. The aim of this study is to assess the potential prognostic value of serum albumin in patients with head and neck adenoid-cystic carcinomas. Patients and Methods: This retrospective cohort study included all patients treated for a head and neck adenoid-cystic carcinoma between 1993 and 1 June 2019 with available pretherapeutic albumin values and clinical follow-up data. The cohort was stratified into a high and low group according to the median albumin value. The log-rank test was used for comparing overall and disease-free survival. Results: A total of 37 patients with complete follow-up data and available pretreatment albumin values were available. The overall mortality and recurrence rates were 21.6% (n = 8) and 45.9% (n = 17), respectively. Survival was shorter in the low albumin group. In particular, the mean overall survival for the low and high albumin groups were 121.0 months and 142.8 months, respectively. However, the difference was not statistically significant (p = 0.155). A statistically significant difference was observed in context with disease-free survival (45.2 months, 95% confidence interval 31.7–58.8 months vs. 114.8 months, 95% confidence interval 79.3–150.4 months; p = 0.029). Conclusion: Our study suggests a potential prognostic value of serum albumin in patients with a head and neck ACC. A further, external validation of our results is warranted. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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12 pages, 1723 KiB

Open AccessArticle

Liquid Biopsy for EGFR Mutation Analysis in Advanced Non-Small-Cell Lung Cancer Patients: Thoughts Drawn from a Real-Life Experience

by Paola Ulivi, Elisabetta Petracci, Matteo Canale, Ilaria Priano, Laura Capelli, Daniele Calistri, Elisa Chiadini, Paola Cravero, Alice Rossi, Angelo Delmonte, Lucio Crinò and Giuseppe Bronte

Biomedicines 2021, 9(10), 1299; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines9101299 - 23 Sep 2021

Cited by 10 | Viewed by 2029

Abstract

Background: Liquid biopsy analysis for EGFR detection in cell-free DNA (cfDNA) from NSCLC patients has become routine. The aim of this study was to explore its applicability in clinical practice. Methods: We collected data of EGFR-mutated NSCLC patients with liquid biopsy analysis. Data [...] Read more.

Background: Liquid biopsy analysis for EGFR detection in cell-free DNA (cfDNA) from NSCLC patients has become routine. The aim of this study was to explore its applicability in clinical practice. Methods: We collected data of EGFR-mutated NSCLC patients with liquid biopsy analysis. Data included test timing, concomitant tissue re-biopsy, therapy change, histology, stage, smoking habits, gender and age. All analyses were performed via a real-time PCR method to analyze EGFR mutations at exons 18, 19, 20 and 21. Variant allele frequency was performed for patients with available sequential EGFR mutation analysis in cfDNA. Overall survival was analyzed through the Kaplan–Meier method. We designed flow charts to show the real-life application of liquid biopsy. Results: We found that liquid biopsy is used in treatment-naïve patients as an alternative to EGFR detection in tumor tissue, and in patients with positive or negative EGFR from tumor biopsy. The majority of liquid biopsy analyses were performed in NSCLC patients who were disease progressive during TKI therapy. The presence of EGFR mutation in cfDNA was associated with a worse prognosis. In two patients, VAF of EGFR mutations in cfDNA was concordant with tumor volume changes. Conclusion: These findings suggest that liquid biopsy for EGFR detection can continue to be useful. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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12 pages, 2338 KiB

Open AccessEditor’s ChoiceArticle

ExoCAS-2: Rapid and Pure Isolation of Exosomes by Anionic Exchange Using Magnetic Beads

by Hyunsung Kim and Sehyun Shin

Biomedicines 2021, 9(1), 28; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines9010028 - 02 Jan 2021

Cited by 27 | Viewed by 9460

Abstract

Extracellular vesicles (EVs) are considered essential biomarkers in liquid biopsies. Despite intensive efforts aimed at employing EVs in a clinical setting, workable approaches are currently limited owing to the fact that EV-isolation technologies are still in a nascent stage. This study introduces a [...] Read more.

Extracellular vesicles (EVs) are considered essential biomarkers in liquid biopsies. Despite intensive efforts aimed at employing EVs in a clinical setting, workable approaches are currently limited owing to the fact that EV-isolation technologies are still in a nascent stage. This study introduces a magnetic bead-based ion exchange platform for isolating EVs called ExoCAS-2 (exosome clustering and scattering). Owing to their negative charge, exosomes can easily adhere to magnetic beads coated with a polycationic polymer. Owing to the features of magnetic beads, exosomes can be easily processed via washing and elution steps and isolated with high purity and yield within 40 min. The present results confirmed the isolation of exosomes through analyses of size distribution, morphology, surface and internal protein markers, and exosomal RNA. Compared with the commercially available methods, the proposed method showed superior performance in terms of key aspects, including operation time, purity, and recovery rate. This highlights the potential of this magnetic bead-based ion exchange platform for isolating exosomes present in blood plasma. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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12 pages, 834 KiB

Open AccessArticle

The Percentage of Free PSA and Urinary Markers Distinguish Prostate Cancer from Benign Hyperplasia and Contribute to a More Accurate Indication for Prostate Biopsy

by Zlata Huskova, Jana Knillova, Zdenek Kolar, Jana Vrbkova, Milan Kral and Jan Bouchal

Biomedicines 2020, 8(6), 173; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines8060173 - 25 Jun 2020

Cited by 6 | Viewed by 2693

Abstract

The main advantage of urinary biomarkers is their noninvasive character and the ability to detect multifocal prostate cancer (CaP). We have previously implemented a quadruplex assay of urinary markers into clinical practice (PCA3, AMACR, TRPM8 and MSMB with KLK3 normalization). In this [...] Read more.

The main advantage of urinary biomarkers is their noninvasive character and the ability to detect multifocal prostate cancer (CaP). We have previously implemented a quadruplex assay of urinary markers into clinical practice (PCA3, AMACR, TRPM8 and MSMB with KLK3 normalization). In this study, we aimed to validate it in a larger cohort with serum PSA 2.5–10 ng/mL and test other selected transcripts and clinical parameters, including the percentage of free prostate-specific antigen (PSA) (% free PSA) and inflammation. In the main cohort of 299 men, we tested the quadruplex transcripts. In a subset of 146 men, we analyzed additional transcripts (CD45, EPCAM, EZH2, Ki67, PA2G4, PSGR, RHOA and TBP). After a prostate massage, the urine was collected, RNA isolated from a cell sediment and qRT-PCR performed. Ct values of KLK3 (i.e., PSA) were strongly correlated with Ct values of other genes which play a role in CaP (i.e., PCA3, AMACR, TRPM8, MSMB and PSGR). AMACR, PCA3, TRPM8 and EZH2 mRNA expression, as well as % free PSA, were significantly different for BPH and CaP. The best combined model (% free PSA plus PCA3 and AMACR) achieved an AUC of 0.728 in the main cohort. In the subset of patients, the best AUC 0.753 was achieved for the combination of PCA3, % free PSA, EPCAM and PSGR. PCA3 mRNA was increased in patients with inflammation, however, this did not affect the stratification of patients indicated for prostate biopsy. In conclusion, the percentage of free PSA and urinary markers contribute to a more accurate indication for prostate biopsy. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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14 pages, 2356 KiB

Open AccessArticle

When Less Is More: Specific Capture and Analysis of Tumor Exosomes in Plasma Increases the Sensitivity of Liquid Biopsy for Comprehensive Detection of Multiple Androgen Receptor Phenotypes in Advanced Prostate Cancer Patients

by Chiara Foroni, Natasa Zarovni, Laura Bianciardi, Simona Bernardi, Luca Triggiani, Davide Zocco, Marta Venturella, Antonio Chiesi, Francesca Valcamonico and Alfredo Berruti

Biomedicines 2020, 8(5), 131; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines8050131 - 22 May 2020

Cited by 33 | Viewed by 3709

Abstract

We evaluated the advantages and the reliability of novel protocols for the enrichment of tumor extracellular vesicles (EVs), enabling a blood-based test for the noninvasive parallel profiling of multiple androgen receptor (AR) gene alterations. Three clinically relevant AR variants related to response/resistance [...] Read more.

We evaluated the advantages and the reliability of novel protocols for the enrichment of tumor extracellular vesicles (EVs), enabling a blood-based test for the noninvasive parallel profiling of multiple androgen receptor (AR) gene alterations. Three clinically relevant AR variants related to response/resistance to standard-of-care treatments (AR-V7 transcript, AR T878A point mutation and AR gene amplification) were evaluated by digital PCR in 15 samples from patients affected by Castration-Resistant Prostate Cancer (CRPC). Plasma was processed to obtain circulating RNA and DNA using protocols based on tumor EVs enrichment through immuno-affinity and peptide-affinity compared to generic extraction kits. Our results showed that immuno-affinity enrichment prior to RNA extraction clearly outperforms the generic isolation method in the detection of AR-V7, also allowing for a distinction between responder (R) and non-responder (NR) patients. The T878A mutation was detected, overall, in nine out of 15 samples and no approach alone was able to reveal mutations in all harboring samples, showing that the employed methods complement each other. AR amplification was detected in the majority of CRPC samples analysed using either cell-free DNA (cfDNA) or exosome isolation kits (80%). We demonstrated that selective isolation of a subset of circulating exosomes enriched for tumor origin, rather than analysis of total plasma exosomes, or total plasma nucleic acids, increases sensitivity and specificity for the detection of specific alterations. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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Review

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20 pages, 1230 KiB

Open AccessReview

Clinical Utility of Liquid Biopsy-Based Actionable Mutations Detected via ddPCR

by Irina Palacín-Aliana, Noemí García-Romero, Adrià Asensi-Puig, Josefa Carrión-Navarro, Víctor González-Rumayor and Ángel Ayuso-Sacido

Biomedicines 2021, 9(8), 906; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines9080906 - 28 Jul 2021

Cited by 26 | Viewed by 9364

Abstract

Cancer is one of the leading causes of death worldwide and remains a major public health challenge. The introduction of more sensitive and powerful technologies has permitted the appearance of new tumor-specific molecular aberrations with a significant cancer management improvement. Therefore, molecular pathology [...] Read more.

Cancer is one of the leading causes of death worldwide and remains a major public health challenge. The introduction of more sensitive and powerful technologies has permitted the appearance of new tumor-specific molecular aberrations with a significant cancer management improvement. Therefore, molecular pathology profiling has become fundamental not only to guide tumor diagnosis and prognosis but also to assist with therapeutic decisions in daily practice. Although tumor biopsies continue to be mandatory in cancer diagnosis and classification, several studies have demonstrated that liquid biopsies could be used as a potential tool for the detection of cancer-specific biomarkers. One of the main advantages is that circulating free DNA (cfDNA) provides information about intra-tumoral heterogeneity, reflecting dynamic changes in tumor burden. This minimally invasive tool has become an accurate and reliable instrument for monitoring cancer genetics. However, implementing liquid biopsies across the clinical practice is still ongoing. The main challenge is to detect genomic alterations at low allele fractions. Droplet digital PCR (ddPCR) is a powerful approach that can overcome this issue due to its high sensitivity and specificity. Here we explore the real-world clinical utility of the liquid biopsy ddPCR assays in the most diagnosed cancer subtypes. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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22 pages, 1774 KiB

Open AccessReview

Current Status and Future Perspectives of Liquid Biopsy in Small Cell Lung Cancer

by Patricia Mondelo-Macía, Jorge García-González, Luis León-Mateos, Adrián Castillo-García, Rafael López-López, Laura Muinelo-Romay and Roberto Díaz-Peña

Biomedicines 2021, 9(1), 48; https://0-doi-org.brum.beds.ac.uk/10.3390/biomedicines9010048 - 07 Jan 2021

Cited by 17 | Viewed by 4767

Abstract

Approximately 19% of all cancer-related deaths are due to lung cancer, which is the leading cause of mortality worldwide. Small cell lung cancer (SCLC) affects approximately 15% of patients diagnosed with lung cancer. SCLC is characterized by aggressiveness; the majority of SCLC patients [...] Read more.

Approximately 19% of all cancer-related deaths are due to lung cancer, which is the leading cause of mortality worldwide. Small cell lung cancer (SCLC) affects approximately 15% of patients diagnosed with lung cancer. SCLC is characterized by aggressiveness; the majority of SCLC patients present with metastatic disease, and less than 5% of patients are alive at 5 years. The gold standard of SCLC treatment is platinum and etoposide-based chemotherapy; however, its effects are short. In recent years, treatment for SCLC has changed; new drugs have been approved, and new biomarkers are needed for treatment selection. Liquid biopsy is a non-invasive, rapid, repeated and alternative tool to the traditional tumor biopsy that could allow the most personalized medicine into the management of SCLC patients. Circulating tumor cells (CTCs) and cell-free DNA (cfDNA) are the most commonly used liquid biopsy biomarkers. Some studies have reported the prognostic factors of CTCs and cfDNA in SCLC patients, independent of the stage. In this review, we summarize the recent SCLC studies of CTCs, cfDNA and other liquid biopsy biomarkers, and we discuss the future utility of liquid biopsy in the clinical management of SCLC. Full article

(This article belongs to the Special Issue Liquid Biopsies in Cancer Diagnosis, Monitoring and Prognosis)

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