Droplet digital PCR improves urinary exosomal miRNA detection compared to real-time PCR

Wang, Cong; Ding, Qiang; Plant, Pamela; Basheer, Mayada; Yang, Chuance; Tawedrous, Eriny; Krizova, Adriana; Boulos, Carl; Farag, Mina; Cheng, Yufeng; Yousef, George M.

doi:10.1016/j.clinbiochem.2019.03.008

Cited by 70 publications

(50 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MiRNAs are small non-coding RNAs responsible for silencing gene expression. They are roughly 22 base pairs long and originate in the nucleus of the cell, but can also be found in the extracellular environment in various biological fluids including serum [21], interstitial fluid [22][23][24], and urine [25][26][27]. More recently, circulating miRNAs have been utilized as biomarkers of environmental and toxicological exposure [21,28], with documented associations between metal exposure and circulating miRNAs [29][30][31].…”

Section: Introductionmentioning

confidence: 99%

Serum-borne factors alter cerebrovascular endothelial microRNA expression following particulate matter exposure near an abandoned uranium mine on the Navajo Nation

et al. 2020

View full text Add to dashboard Cite

Background: Commercial uranium mining on the Navajo Nation has subjected communities on tribal lands in the Southwestern United States to exposures from residual environmental contamination. Vascular health effects from these ongoing exposures are an active area of study. There is an association between residential mine-site proximity and circulating biomarkers in residents, however, the contribution of mine-site derived wind-blown dusts on vascular and other health outcomes is unknown. To assess neurovascular effects of mine-site derived dusts, we exposed mice using a novel exposure paradigm, the AirCARE1 mobile inhalation laboratory, located 2 km from an abandoned uranium mine, Claim 28 in Blue Gap Tachee, AZ. Mice were exposed to filtered air (FA) (n = 6) or concentrated ambient particulate matter (CAPs) (n = 5) for 2 wks for 4 h per day. Results: To assess miRNA differential expression in cultured mouse cerebrovascular cells following particulate matter (PM) exposure (average: 96.6 ± 60.4 μg/m 3 for all 4 h exposures), the serum cumulative inflammatory potential (SCIP) assay was employed. MiRNA sequencing was then performed in cultured mouse cerebrovascular endothelial cells (mCECs) to evaluate transcriptional changes. Results indicated 27 highly differentially expressed (p < 0.01) murine miRNAs, as measured in the SCIP assay. Gene ontology (GO) pathway analysis revealed notable alterations in GO enrichment related to the cytoplasm, protein binding and the cytosol, while significant KEGG pathways involved pathways in cancer, axon guidance and Wnt signaling. Expression of these 27 identified, differentially expressed murine miRNAs were then evaluated in the serum. Nine of these miRNAs (~30%) were significantly altered in the serum and 8 of those miRNAs demonstrated the same directional change (either upregulation or downregulation) as cellular miRNAs, as measured in the SCIP assay. Significantly upregulated miRNAs in the CAPs exposure group included miRNAs in the let-7a family. Overexpression of mmu-let-7a via transfection experiments, suggested that this miRNA may mediate mCEC barrier integrity following dust exposure.

show abstract

Section: Introductionmentioning

confidence: 99%

Serum-borne factors alter cerebrovascular endothelial microRNA expression following particulate matter exposure near an abandoned uranium mine on the Navajo Nation

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Moreover, the choice of assay can change the sensitivity of the results; for instance, there is a tradeoff between using microarray characterization of miRNA in EVs with lower cost versus RNA sequencing with higher cost and broader dynamic range [ 102 ]. In fact, using high accuracy platforms such as digital droplet PCR showed increased assay accuracy in the analysis of EV-based nucleic acids [ 103 ]. Vortexing the biofluid after thawing increased the EV recovery [ 104 ].…”

Section: Challenges For Using Evs As Biomarkers: Isolation Clinicmentioning

confidence: 99%

Extracellular Vesicles as Biomarkers in Cancer Immunotherapy

Mathew

Zade

Mezghani

et al. 2020

Cancers

View full text Add to dashboard Cite

Extracellular vesicles (EVs), including exosomes and microvesicles, are membrane-bound vesicles secreted by most cell types during both physiologic conditions as well in response to cellular stress. EVs play an important role in intercellular communication and are emerging as key players in tumor immunology. Tumor-derived EVs (TDEs) harbor a diverse array of tumor neoantigens and contain unique molecular signature that is reflective of tumor’s underlying genetic complexity. As such they offer a glimpse into the immune tumor microenvironment (TME) and have the potential to be a novel, minimally invasive biomarker for cancer immunotherapy. Immune checkpoint inhibitors (ICI), such as anti- programmed death-1(PD-1) and its ligand (PD-L1) antibodies, have revolutionized the treatment of a wide variety of solid tumors including head and neck squamous cell carcinoma, urothelial carcinoma, melanoma, non-small cell lung cancer, and others. Typically, an invasive tissue biopsy is required both for histologic diagnosis and next-generation sequencing efforts; the latter have become more widespread in daily clinical practice. There is an unmet need for noninvasive or minimally invasive (e.g., plasma-based) biomarkers both for diagnosis and treatment monitoring. Targeted analysis of EVs in biospecimens, such as plasma and saliva could serve this purpose by potentially obviating the need for tissue sample. In this review, we describe the current challenges of biomarkers in cancer immunotherapy as well as the mechanistic role of TDEs in modulating antitumor immune response

show abstract

“…7 Droplet digital PCR (ddPCR) has emerged as a new generation of PCR technology for quantication of miRNA and circular RNA (circRNA), with improved precision, sensitivity and reproducibility compared to RT-qPCR. 12,13 Nevertheless, requirements of sample purity and quality, thermocycling conditions and relatively high cost of ddPCR limit its application for early detection. A novel cyclic enzymatic repairing-mediated signal amplication has been reported recently for uorescent detection of lncRNA HOTAIR in cancer cells.…”

Section: Introductionmentioning

confidence: 99%

A simple and rapid colorimetric detection of serum lncRNA biomarkers for diagnosis of pancreatic cancer

2020

View full text Add to dashboard Cite

show abstract

Droplet digital PCR improves urinary exosomal miRNA detection compared to real-time PCR

Cited by 70 publications

References 30 publications

Serum-borne factors alter cerebrovascular endothelial microRNA expression following particulate matter exposure near an abandoned uranium mine on the Navajo Nation

Serum-borne factors alter cerebrovascular endothelial microRNA expression following particulate matter exposure near an abandoned uranium mine on the Navajo Nation

Extracellular Vesicles as Biomarkers in Cancer Immunotherapy

A simple and rapid colorimetric detection of serum lncRNA biomarkers for diagnosis of pancreatic cancer

Contact Info

Product

Resources

About