DigiMir Test: Establishing a Novel Pipeline for MiR-371a Quantification Using Droplet Digital PCR in Liquid Biopsies From Testicular Germ Cell Tumor Patients

Sequeira, José Pedro; Lobo, João; Constâncio, Vera; Brito-Rocha, Tiago; Carvalho‐Maia, Carina; Braga, Isaac; Maurício, Joaquina; Henrique, Rui; Jerónimo, Carmen

doi:10.3389/fonc.2022.876732

Cited by 14 publications

(22 citation statements)

References 59 publications

(74 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This result is in line with all previous reports on M371 (Dieckmann et al 2019a;Mørup et al 2020;Sequeira et al 2022;Syring et al 2015;Murray et al 2018;Piao et al 2021) and it clearly underscores the usefulness of the M371 test reiterating the demand for its prompt clinical implementation (Leão et al 2021). Noteworthy, we found a modest but significantly lower expression rate of M371 in seminoma compared to nonseminoma (82.7% versus 93.6%) which had been documented in most of the recent reports (van Agthoven and Looijenga 2017; Dieckmann et al 2019a;Mørup et al 2020) but not in all of them (Myklebust et al 2021;Vilela-Salgueiro et al 2018;Mego et al 2019).…”

Section: Association Of Tumor Marker Expression Rates With Histologysupporting

confidence: 92%

“…The confidence intervals do not overlap, and this result is consistent with the significant difference between the expression rates of M371 and AFP/bHCG in nonseminoma documented with McNemar's test. The very high sensitivity of M371 found herein is consistent with previous studies (Dieckmann et al 2019a;Leão et al 2021;van Agthoven and Looijenga 2017;Mego et al 2019;Mørup et al 2020;Myklebust et al 2021;Lobo et al 2019;Nappi et al 2019;Badia et al 2021;Ye et al 2022;Sequeira et al 2022) underscoring the great superiority of M371 to the classical markers.…”

Section: Association Of Tumor Marker Expression Rates With Clinical S...supporting

confidence: 92%

See 1 more Smart Citation

Testicular neoplasms: the interrelationships of serum levels of microRNA-371a-3p (M371) and classical tumor markers with histology, clinical staging, and age—a statistical analysis

Dieckmann

Dumlupinar

Grobelny

et al. 2023

J Cancer Res Clin Oncol

View full text Add to dashboard Cite

Purpose In testicular neoplasms, the interrelationship of elevations of the novel serum tumor marker microRNA-371a-3p (M371) and traditional markers with other clinical features is still incompletely understood. The present study evaluated marker expression rates in relation to various other clinical parameters. Methods The following data were retrospectively registered from 641 consecutive patients with testicular neoplasms: histology, such as seminoma (n = 365), nonseminoma (n = 179), benign tumor (n = 79), other malignant tumor (n = 18); patients age (years); clinical stage (CS1, CS2a/b, CS2c, CS3); and preoperative elevation of beta HCG, AFP, LDH, M371 (yes/no). Descriptive statistical methods were employed with comparisons of various subgroups to disclose associations of marker expression rates with age, histology and CS, and of age with histology. Results The histologic subgroups revealed significantly different expression rates of tumor markers. M371 performed best with expression rates of 82.69% and 93.58% in seminoma and in nonseminoma, respectively. In germ cell tumors, all markers had significantly higher expression rates in metastasized stages than in localized disease. All markers except LDH have significantly higher expression rates in younger than in older patients. Nonseminoma is most prevalent in the youngest age category, seminoma predominates in patients > 40 years, other malignancies were restricted to patients > 50 years. Conclusion The study documented significant associations of serum marker expression rates with histology, age and clinical staging, with highest rates in nonseminomas, young age and advanced clinical stages. M371 showed significantly higher expression rates than other markers suggesting its superior clinical usefulness.

show abstract

Section: Association Of Tumor Marker Expression Rates With Histologysupporting

confidence: 92%

Section: Association Of Tumor Marker Expression Rates With Clinical S...supporting

confidence: 92%

Testicular neoplasms: the interrelationships of serum levels of microRNA-371a-3p (M371) and classical tumor markers with histology, clinical staging, and age—a statistical analysis

Dieckmann

Dumlupinar

Grobelny

et al. 2023

J Cancer Res Clin Oncol

View full text Add to dashboard Cite

show abstract

“…), method of extraction (beads vs. columns, etc. ), sample quality control measures (hemolysis and extraction efficacy) ( 23 ), and PCR methodology (regular real-time versus digital droplet PCR) ( 24 , 25 ) will also most likely impact optimised protocol development and should be harmonised for creating a universal, robust, clinical test for guiding treatment decisions.…”

Section: Discussionmentioning

confidence: 99%

Impact of differing methodologies for serum miRNA-371a-3p assessment in stage I testicular germ cell cancer recurrence

et al. 2022

Self Cite

View full text Add to dashboard Cite

IntroductionCurrent evidence shows that serum miR-371a-3p can identify disease recurrence in testicular germ cell tumour (TGCT) patients and correlates with tumour load. Despite convincing evidence showing the advantages of including miR-371a-3p testing to complement and overcome the classical serum tumour markers limitations, the successful introduction of a serum miRNA based test into clinical practice has been impeded by a lack of consensus regarding optimal methodologies and lack of a universal protocol and thresholds. Herein, we investigate two quantitative real-time PCR (qRT-PCR) based pipelines in detecting disease recurrence in stage I TGCT patients under active surveillance, and compare the sensitivity and specificity for each method.MethodsSequential serum samples collected from 33 stage I TGCT patients undergoing active surveillance were analysed for miR-371a-3p via qRT-PCR with and without an amplification step included.ResultsUsing a pre-amplified protocol, all known recurrences were detected via elevated miR-371a-3p expression, while without pre-amplification, we failed to detect recurrence in 3/10 known recurrence patients. For pre-amplified analysis, sensitivity and specificity was 90% and 94.4% respectively. Without amplification, sensitivity dropped to 60%, but exhibited 100% specificity.DiscussionWe conclude that incorporating pre-amplification increases sensitivity of miR-371a-3p detection, but produces more false positive results. The ideal protocol for quantification of miR-371a-3p still needs to be determined. TGCT patients undergoing active surveillance may benefit from serum miR-371a-3p quantification with earlier detection of recurrences compared to current standard methods. However, larger cross-institutional studies where samples are processed and data is analysed in a standardised manner are required prior to its routine clinical implementation.

show abstract

“…In a recent study, miR‐371a‐3p expression was assessed using two different methods: qualitative polymerase chain reaction (qPCR) and digital‐droplet PCR (ddPCR) 53 . The utility of the ddPCR was confirmed in a recent study of 107 TGCT samples where miR‐371a‐3p achieved 100% specificity with absolute quantification, mirroring tumour burden 54 …”

Section: Cs‐ii/iii Monitoring Treatment Responsementioning

confidence: 99%

miRNAs for testicular germ cell tumours: Contemporary indications for diagnosis, surveillance and follow‐up

Chavarriaga

Hamilton

2022

Andrology

View full text Add to dashboard Cite

Despite excellent outcomes with modern multidisciplinary care, clinicians caring for patients with testicular germ cell tumour (TGCTs) face clinical dilemmas across the spectrum of disease. Wrong treatment choices can lead to undertreatment or overtreatment of these young men. Unfortunately the currently available biomarkers alpha‐fetoprotein and human chorionic gonadotropin, lack sufficient sensitivity and specificity to reliably aid in these clinical dilemmas. Thus, a sensitive and specific biomarker is desperately needed. Serum or plasma miRNA, in particular, miR‐371a‐3p, has shown great promise in discriminating the presence of TGCT and may represent a breakthrough for this disease. In this review, we discuss the potential role of miRNA across clinical states of TGCTs. We review their discovery, methods of assay, limitations and future potential.

show abstract

DigiMir Test: Establishing a Novel Pipeline for MiR-371a Quantification Using Droplet Digital PCR in Liquid Biopsies From Testicular Germ Cell Tumor Patients

Cited by 14 publications

References 59 publications

Testicular neoplasms: the interrelationships of serum levels of microRNA-371a-3p (M371) and classical tumor markers with histology, clinical staging, and age—a statistical analysis

Testicular neoplasms: the interrelationships of serum levels of microRNA-371a-3p (M371) and classical tumor markers with histology, clinical staging, and age—a statistical analysis

Impact of differing methodologies for serum miRNA-371a-3p assessment in stage I testicular germ cell cancer recurrence

miRNAs for testicular germ cell tumours: Contemporary indications for diagnosis, surveillance and follow‐up

Contact Info

Product

Resources

About