Evaluation of micro<scp>RNA</scp> stability in feces from healthy dogs

Cirera, Susanna; Willumsen, Line Møller; Johansen, Thea T.; Nielsen, Lise Nikolic

doi:10.1111/vcp.12566

Cited by 6 publications

(12 citation statements)

References 29 publications

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“…No statistically significant difference between miRNA expression in samples stored for 24 hours at RT, 24 hours at 4°C, or directly at −20°C were identified. These results are in contrast to prior findings in feces from healthy dogs, where a significant difference was found in cfa‐miR‐16 and cfa‐miR‐21 stored at RT and −20°C, respectively . A sample (RT) from one cat was excluded because 8/10 data points were missing or Cq values were below the lower limit of quantitation.…”

Section: Discussioncontrasting

confidence: 92%

“…This study is the first to identify fecal miRNAs in cats. Some of these miRNAs have been previously isolated in blood or tissue from cats or dogs, while others have been implicated in tumorigenesis in humans and mice, which makes them interesting targets for future analyses in cats with GI cancer.…”

Section: Discussionmentioning

confidence: 99%

“…Recently fecal miRNA has been investigated as a noninvasive screening test for colorectal cancer (CRC) in humans . Our group has validated an assay for extraction of fecal miRNA from dogs and assessed its stability; however, no similar information exists in cats …”

Section: Introductionmentioning

confidence: 99%

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Evaluation of fecal microRNA stability in healthy cats

Lyngby

Kristensen

Fredholm

et al. 2019

Veterinary Clinical Pathol

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BackgroundGastrointestinal (GI) cancer accounts for 14% of feline malignancies. There is a great need for reliable noninvasive diagnostic biomarkers to reach a timely diagnosis and initiate treatment. Fecal microRNAs (miRNAs) could be such a biomarker and have shown great potential in colorectal screening in people but have yet to be investigated in cats.ObjectivesWe aimed to evaluate the presence and stability of feline fecal miRNA under different storage conditions (room temperature [RT], 4, and −20°C) and to evaluate the expression levels of specific fecal miRNAs collected on three separate days (days 1, 4, and 7) in healthy cats.MethodsHealthy cats were prospectively recruited. Fecal samples were collected, aliquoted, and stored for 24 hours at RT and then transferred to −20°C, stored for 24 hours at 4°C and then transferred to −20°C, or were immediately placed at −20°C on day 1 or at −20°C on days 4 and 7 postcollection. Expression of 22 miRNAs was investigated using quantitative real‐time PCR.ResultsTen miRNA assays worked well, and one, let‐7b, was used for normalization. No differences in miRNA expression were seen between the three storage temperatures for the nine miRNAs investigated. Only miR‐26a showed a significant increase in expression between samples of days 1 and 7. The rest of the miRNAs levels were stable over time.ConclusionsFecal miRNA can be isolated from healthy cats. The expression was stable at different temperatures and for most of the miRNAs over time. Prospective studies evaluating fecal miRNA as biomarkers in cats with GI neoplasia are warranted.

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Section: Discussioncontrasting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

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Evaluation of fecal microRNA stability in healthy cats

Lyngby

Kristensen

Fredholm

et al. 2019

Veterinary Clinical Pathol

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“…A number of biomarkers for GIC have been investigated in human and veterinary medicine 5‐10 . MicroRNAs (miRNAs) have recently been proposed as promising diagnostic and prognostic markers of GI disease in both humans and dogs 11‐16 . MicroRNAs are small, non–protein‐coding RNA molecules of approximately 20 to 23 nucleotide length, which primarily suppress the expression of target genes by binding to messenger RNA (mRNA) and inhibiting translation 17 .…”

Section: Introductionmentioning

confidence: 99%

Association of fecal and serum microRNA profiles with gastrointestinal cancer and chronic inflammatory enteropathy in dogs

Lyngby

Gòdia

Brogaard

et al. 2022

Veterinary Internal Medicne

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Background: Reliable biomarkers to differentiate gastrointestinal cancer (GIC) from chronic inflammatory enteropathy (CIE) in dogs are needed. Fecal and serum micro-RNAs (miRNAs) have been proposed as diagnostic and prognostic markers of GI disease in humans and dogs.Hypothesis/Objectives: Dogs with GIC have fecal and serum miRNA profiles that differ from those of dogs with CIE. Aims: (a) identify miRNAs that differentiate GIC from CIE, (b) use high-throughput reverse transcription quantitative real-time PCR (RT-qPCR) to establish fecal and serum miRNA panels to distinguish GIC from CIE in dogs.

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“…For each miRNA assay, primers were designed using a publicly available software “miRprimer” (Table S2). Quantitative PCR was performed on Mx3005P qPCR system (Sensititre Thermo Scientific Fisher), with qPCR reactions, conditions, melting curve, and PCR efficiency according to Cirera et al Quantitative PCR data were pre‐processed using GenEx 6 Pro program (GenEx 6 Pro program, MultiD, Gothenburg, Sweden). Briefly: (1) data were efficiency corrected; (2) data were corrected to the spike‐in miRNA for the stability study (I and II) and normalized to miR23a (stable reference miRNA selected by NormFinder and GeNorm programs for the clinical study); (3) both replicates were included in the linear mixed‐effects models (LMMs) in the stability study part I and II, while an average was used for the clinical study; (4) relative quantities (fold changes) were calculated in relation to the sample showing the lowest expression (higher Cq) for the stability study (I and II), or in relation to the average of the control group for the clinical study; and (5) data were log2 transformed before proceeding to statistical analysis.…”

Section: Methodsmentioning

confidence: 99%

Stability and profiling of urinary microRNAs in healthy cats and cats with pyelonephritis or other urological conditions

Jessen

Nielsen

Kieler

et al. 2019

Veterinary Internal Medicne

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Background: Specific biomarkers of pyelonephritis (PN) in cats are lacking. Micro-RNAs (miRNAs) have diagnostic potential in human nephropathies.Objectives: To investigate the presence/stability of miRNAs in whole urine of cats and the discriminatory potential of selected urinary miRNAs for PN in cats.Animals: Twelve healthy cats, 5 cats with PN, and 13 cats with chronic kidney disease (n = 5), subclinical bacteriuria (n = 3), and ureteral obstructions (n = 5) recruited from 2 companion animal hospitals.Methods: Prospective case-control study. Expression profiles of 24 miRNAs were performed by quantitative PCR (qPCR). Effect of storage temperature (4 C [24 hours], −20 C, and −80 C) was determined for a subset of miRNAs in healthy cats.Results: Urinary miR-4286, miR-30c, miR-204, miR4454, miR-21, miR-16, miR-191, and miR-30a were detected. For the majority of miRNAs tested, storage at 4 C and −20 C resulted in significantly lower miRNA yield compared to storage at −80 C (mean log2fold changes across miRNAs from −0.5 ± 0.4 SD to −1.20 ± 0.4 SD (4 C versus −80 C) and from −0.7 ± 0.2 SD to −1.20 ± 0.3 SD (−20 C versus −80 C)).Cats with PN had significantly upregulated miR-16 with a mean log2fold change of 1.0 ± 0.4 SD, compared with controls (−0.1 ± 0.2, P = .01) and other urological conditions (0.6 ± 0.3, P = .04).Conclusions: Upregulation of miR16 might be PN-specific, pathogen-specific (Escherichia coli), or both. K E Y W O R D SBiomarker, CKD, feline, pathogen-directed microRNA expression, subclinical bacteriuria, ureteral obstruction, UTI

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Evaluation of microRNA stability in feces from healthy dogs

Cited by 6 publications

References 29 publications

Evaluation of fecal microRNA stability in healthy cats

Evaluation of fecal microRNA stability in healthy cats

Association of fecal and serum microRNA profiles with gastrointestinal cancer and chronic inflammatory enteropathy in dogs

Stability and profiling of urinary microRNAs in healthy cats and cats with pyelonephritis or other urological conditions

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