Recent Advances in Exosomal miRNA Biosensing for Liquid Biopsy

Lin, Bingqian; Jiang, Jinting; Jia, Jie–Min; Zhou, Xiang

doi:10.3390/molecules27217145

Cited by 17 publications

(15 citation statements)

References 91 publications

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“…Exosomal miRNAs offer a great deal of promise, however, there are still certain challenges to be resolved. For example, the detection of exosomal miRNAs should be standardized [ 107 ]. Until now, most exosomal miRNAs-associated studies have only been conducted with just a small number of clinical samples used, and experiment settings and detection criteria vary from lab to lab [ 107 ].…”

Section: Challenges In Clinical Applications Of Exosomal Micrornasmentioning

confidence: 99%

“…For example, the detection of exosomal miRNAs should be standardized [ 107 ]. Until now, most exosomal miRNAs-associated studies have only been conducted with just a small number of clinical samples used, and experiment settings and detection criteria vary from lab to lab [ 107 ]. Additionally, no standardized techniques exist for collecting exosomes, deconstructing them, extracting, and storing miRNA [ 107 ].…”

Section: Challenges In Clinical Applications Of Exosomal Micrornasmentioning

confidence: 99%

“…Until now, most exosomal miRNAs-associated studies have only been conducted with just a small number of clinical samples used, and experiment settings and detection criteria vary from lab to lab [ 107 ]. Additionally, no standardized techniques exist for collecting exosomes, deconstructing them, extracting, and storing miRNA [ 107 ]. Furthermore, the vast majority of investigations focused on exosomal miRNA levels in serum and plasma.…”

Section: Challenges In Clinical Applications Of Exosomal Micrornasmentioning

confidence: 99%

See 2 more Smart Citations

Emerging functions and clinical applications of exosomal microRNAs in diseases

Ghafouri‐Fard

Shoorei

Dong

et al. 2023

Non-coding RNA Research

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Section: Challenges In Clinical Applications Of Exosomal Micrornasmentioning

confidence: 99%

Section: Challenges In Clinical Applications Of Exosomal Micrornasmentioning

confidence: 99%

Section: Challenges In Clinical Applications Of Exosomal Micrornasmentioning

confidence: 99%

See 1 more Smart Citation

Emerging functions and clinical applications of exosomal microRNAs in diseases

Ghafouri‐Fard

Shoorei

Dong

et al. 2023

Non-coding RNA Research

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“…Exosomes have emerged as a promising target for liquid biopsy, thanks to their abundance in body fluids, stable circulation, and specific bioactive cargo. − Of particular interest are microRNAs (miRNAs) found within exosomes, which are derived from donor cells and act on recipient cells as essential gene expression regulators, − mediating intercellular communication and influencing cancer development. − MiRNAs within exosomes have demonstrated superior sensitivity and specificity compared to single cancer protein markers, making them valuable tools for early cancer screening, predicting cancer tumor, node, metastasis (TNM) staging, assessing prognosis, monitoring therapeutic effects, and evaluating drug resistance. − Therefore, the detection of exosomal miRNAs offers the potential for accurate and noninvasive diagnosis of patients, aligning with the trend of precision medicine and holding promising prospects for widespread clinical applications. − …”

Section: Introductionmentioning

confidence: 99%

Dumbbell Dual-Hairpin Triggered DNA Nanonet Assembly for Cascade-Amplified Sensing of Exosomal MicroRNA

Li,

Tang,

Deng

et al. 2024

ACS Omega

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Exosomal microRNAs (miRNAs) are valuable biomarkers closely associated with cancer progression. Therefore, sensitive and specific exosomal miRNA biosensing has been employed for cancer diagnosis, prognosis, and prediction. In this study, a miRNA-based DNA nanonet assembly strategy is proposed, enabling the biosensing of exosomal miRNAs through dumbbell dual-hairpin under isothermal enzyme-free conditions. This strategy dexterously designs a specific dumbbell dual-hairpin that can selectively recognize exosomal miRNA, inducing conformational changes to cascade-generated X-shaped DNA structures, facilitating the extension of the X-shaped DNA in three-dimensional space, ultimately forming a DNA nanonet assembly. On the basis of the target miRNA, our design enriches the fluorescence signal through the cascade assembly of DNA nanonet and realizes the secondary signal amplification. Using exosomal miR-141 as the target, the resultant fluorescence sensing demonstrates an impressive detection limit of 57.6 pM and could identify miRNA sequences with single-base variants with high specificity. Through the analysis of plasma and urine samples, this method effectively distinguishes between benign prostatic hyperplasia, prostate cancer, and metastatic prostate cancer. Serving as a novel noninvasive and accurate screening and diagnostic tool for prostate cancer, this dumbbell dual-hairpin triggered DNA nanonet assembly strategy is promising for clinical applications.

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“…A particular subset of EVs, namely exosomes, are particularly of interest due to their role in cell-cell communication, tumorigenesis, protecting the nucleic acid cargo from degradation and the tumour specificity of secreted EVs. Furthermore, the recent focus on EV specific miRNAs is due to their involvement in cell proliferation, invasion, apoptosis, angiogenesis and EMT, all hallmarks of cancer development [641][642][643]. Moreover, miRNAs have recently been identified as a novel class of tumour suppressors or promoters, with the latter termed 'oncomiRs' [644].…”

Section: Ev Mirnas As Biomarkers In a Clinical Settingmentioning

confidence: 99%

Identifying Novel Candidate Biomarkers of Early Epithelial Ovarian Cancer

Sczelecki¹

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Ovarian cancer (OC) is the fifth leading cause of cancer deaths in women, and is the most lethal gynaecological cancer worldwide, with the most common subtype being epithelial OC (EOC). The asymptomatic nature and limited understanding of early disease hampers research into early-stage OC and culminates in diagnostic methods only capable of detecting malignant markers of OC. Therefore, there is an urgent need for biomarkers of early-stage OC to be identified. The first aim of this thesis was to validate a unique mouse model of early OC development. The homozygous Fanconi anemia complementation group D2 knock-out mouse (Fancd2-/-) develops multiple ovarian tumor phenotypes in a sequential manner as they age. Our group previously identified by immunohistochemistry the purported initiating precursor cells, termed ‘sex cords’, that are hypothesised to progress into EOC in this model. To genetically validate this hypothesis, the sex cords, a late-stage tumor phenotype (tubulostromal adenoma), and Fancd2 wild-type (Fancd2+/+) granulosa cell and stromal controls, were isolated by laser-capture microdissection for downstream genetic investigation. Multiplexed gene expression analyses of ~ 90 selected genes were performed using the Genome Lab GeXP Genetic Analysis System (GeXP). Principal component analyses and unbiased hierarchical clustering of the resultant gene expression data determined that the gene signature profiles within sex cords and tubulostromal adenoma samples indeed clustered together genetically confirming the identity of a precursor lesion in this mouse model. Furthermore, key epithelial OC markers included in the GeXP assay (Pax8, Cdh1 and Muc16) were expressed in both the sex cords and tubulostromal adenoma demonstrating neoplastic transformation was already occurring in the precursor lesion. Despite confirmation of the identity and developmental origin of the precursor lesion (sex cords), an in-depth understanding of the model was limited due to restricted numbers of genes able to be investigated using the GeXP assay, thus unique markers of the precursor lesion were not identified. Therefore, the second aim of this thesis was to comprehensively investigate the genetic signature of the Fancd2-/- model via total and small RNA sequencing of micro-dissected cells of the sex cords, tubulostromal adenomas and the equivalent Fancd2+/+ controls. Sequencing results of matched messenger (m) RNAs and micro (mi) RNA expression in micro-dissected tissue of the phenotypes of interest provided novel information about the development of early EOC and unique transcriptional markers, namely upregulated Brinp2, Slc6a7, Smpd3, Grin2b, Grik4, AW55198, Lrp11 and Mro mRNAs and downregulated miR-122/b-5p, miR-129-5p, miR-138-5p, miR-465a/b/-3p, miR-465c-3p/5p, miR-500-3p and miR-129b-3p miRNAs of the sex cord phenotype through differential gene expression analysis. Whilst tissue markers of EOC are relevant as biomarkers of EOC, the location of the ovaries within the peritoneum prevents their diagnostic implementation without surgical intervention. To circumvent this limitation, liquid biopsy has become paramount for OC diagnosis. However, the detection of early biomarkers of EOC in humans is ultimately hampered by their low systemic concentration and the presence of circulating factors from all other organ systems. Therefore, to bypass detection limitations within systemic circulation, the next aim of this thesis was to leverage the established Fancd2-/- model of early EOC to provide proof of concept that secreted miRNA encapsulated into extracellular vesicles (EV) secreted from the precursor lesion (i.e., sex cords) could be detected in conditioned media from ovary explant culture. Performing RNA sequencing from EV-derived small RNAs isolated from conditioned culture medium resulted in the identification of five common upregulated secreted EV miRNAs in Fancd2-/- mouse ovaries that had sex cords or tubulostromal adenomas present, compared to their age-matched Fancd2+/+ counterparts. Additionally, the small RNA sequencing data of EVs was compared to the sequencing data of the micro-dissected sex cords and tubulostromal adenomas and these identical five miRNAs were also significantly upregulated as compared to the control group. This suggests that these five secreted EV miRNAs (miR-200a-3p, miR-200b-3p, miR-200c-3p, miR-429-3p and miR-375-3p) originate from the sex cord and tubulostromal adenoma cells and therefore provided proof-of-concept that small transformational changes such as the development of a precursor lesion can be detected within secreted EVs. The final aim of this thesis was to determine whether the methodologies adopted for early candidate biomarker discovery in a murine model can be applied to human biopsy specimens and subsequently the relevance of the Fancd2-/- model for identifying novel biomarkers for human EOC. Two serous subtypes of human EOC, early low-grade serous borderline tumours (SBT; as an early phenotype) and high-grade serous ovarian cancer (HGSOC; as a late malignant phenotype) were selected for study. SBT and HGSOC specimens were prepared for matched micro-dissected neoplastic epithelial cells and explant cultures. Total and small RNA sequencing of micro-dissected tissue and small RNA sequencing of EVs from explant cultures were performed as per the mouse studies. This sequencing data resulted in the identification of SBT tissue-specific mRNA and miRNA markers as well as SBT-secreted miRNA markers. Remarkably, comparing the human and mouse sequencing data revealed an overlap in differentially expressed genes between the SBT, HGSOC, sex cords, and tubulostromal adenomas, pertaining to key epithelial markers of EOC (Muc16, Cdh1, Krt7, Krt19, Epcam) and the five secreted EV miRNAs firstly described in the murine studies. This provides strong evidence that these novel candidate biomarkers are relevant to early human EOC. Overall, the results from this thesis confirmed the relevance of the Fancd2 model for human OC investigation and identified novel candidate biomarkers of early EOC that may be applied to both the mouse model and human ovarian cancer.

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Recent Advances in Exosomal miRNA Biosensing for Liquid Biopsy

Cited by 17 publications

References 91 publications

Emerging functions and clinical applications of exosomal microRNAs in diseases

Emerging functions and clinical applications of exosomal microRNAs in diseases

Dumbbell Dual-Hairpin Triggered DNA Nanonet Assembly for Cascade-Amplified Sensing of Exosomal MicroRNA

Identifying Novel Candidate Biomarkers of Early Epithelial Ovarian Cancer

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