Droplet digital PCR for quantification of<i>ITGA6</i>in a stool mRNA assay for the detection of colorectal cancers

Herring, Elizabeth; Kanaoka, Shigeru; Tremblay, Éric; Beaulieu, Jean‐François

doi:10.3748/wjg.v23.i16.2891

Cited by 16 publications

(16 citation statements)

References 35 publications

(66 reference statements)

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“…Indeed, ITGA6 levels were found to be increased in stool samples of AA and CRC at all stages while receiver operating characteristic (ROC) curves revealed that ITGA6 as a single marker can predict 75% of the AA and 81% of the CRC, with a 88% specificity [69]. Comparable results were also obtained using droplet digital PCR [70]. Interestingly, increases in stool ITGA6A levels were only found in samples from patients with CRC, consistent with previous observations in primary tumours [43,44].…”

Section: Screening Factorssupporting

confidence: 87%

Integrin α6β4 in Colorectal Cancer: Expression, Regulation, Functional Alterations and Use as a Biomarker

Beaulieu

2019

Cancers

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Integrin α6β4 is one of the main laminin receptors and is primarily expressed by epithelial cells as an active component of hemidesmosomes. In this article, after a brief summary about integrins in the gut epithelium in general, I review the knowledge and clinical potential of this receptor in human colorectal cancer (CRC) cells. Most CRC cells overexpress both α6 and β4 subunits, in situ in primary tumours as well as in established CRC cell lines. The mechanisms that lead to overexpression have not yet been elucidated but clearly involve specific transcription factors such as MYC. From a functional point of view, one key element affecting CRC cell behaviour is the relocalization of α6β4 to the actin cytoskeleton, favouring a more migratory and anoikis-resistant phenotype. Another major element is its expression under various molecular forms that have the distinct ability to interact with ligands (α6β4 ± ctd) or to promote pro- or anti-proliferative properties (α6Aβ4 vs. α6Bβ4). The integrin α6β4 is thus involved in most steps susceptible to participation with CRC progression. The potential clinical significance of this integrin has begun to be investigated and recent studies have shown that ITGA6 and ITGB4 can be useful biomarkers for CRC early detection in a non-invasive assay and as a prognostic factor, respectively.

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Section: Screening Factorssupporting

confidence: 87%

Integrin α6β4 in Colorectal Cancer: Expression, Regulation, Functional Alterations and Use as a Biomarker

Beaulieu

2019

Cancers

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“…Given these advantages, it can be used to determine the expression and copy number variation analysis of the target gene [19,20] .…”

Section: Discussionmentioning

confidence: 99%

Development of a reliable method to diagnose Streptococcus agalactiae infection by droplet digital PCR

Guo¹,

Zeng

Chen

et al. 2020

Preprint

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Background: Streptococcus agalactiae (GBS) is the causative pathogen of puerperal sepsis in pregnant women and meningitis in infants. Infection of GBS is responsible for the increased morbidity in pregnant women and the elderly, and bring challenges to clinical diagnosis and treatment. However, culture-based approaches to detect S.agalactiae is time-consuming with limited sensitivity. Besides, real-time quantitative PCR demand for expensive instruments with tedious steps. Thus, we aim to establish a new detection method for more accurate and rapid detection of S.agalactiae.Results: The ddPCR primer targeted the CspE gene showed a better amplified efficiency in the reaction. The limitation of ddPCR for identifying GBS DNA was able to reach 5 pg/µL. Moreover, no positive amplified signals could be detected in the reactions which served 11 non-GBS strains DNA as templates. Furthermore, the coefficient of variation of this method is 4.5%, indicating an excellent repeatability of ddPCR assay.Conclusions: In our study, ddPCR was performed as a rapid detection of S.agalactiae with high sensitivity and specificity. This technique can promote the accuracy of the diagnosis of GBS infection and provide a scientific basis for clinical treatment.

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“…Interestingly, in a very recent study, ddPCR was also used to quantify an mRNA biomarker, ITGA6 , in stool from patients with CRC[ 17 ]. Tumor-derived nucleic acids present in stool samples come from the exfoliation of tumor cells of the intestinal mucosa and are a non-invasive, alternative source of genetic material for KRAS oncogene mutation screening in CRC patients[ 33 ].…”

Section: Discussionmentioning

confidence: 99%

“…Stool-derived DNA is a potential non-invasive alternative source of DNA for tumor genotyping in CRC due to the high rate of exfoliation of tumor cells into the bowel lumen[ 17 ]. Digital PCR was first described in 1999 by Vogelstein and Kinzler in a study aimed at identifying KRAS in DNA obtained from fecal samples of CRC patients[ 18 ].…”

Section: Introductionmentioning

confidence: 99%

“…The introduction of new instrumentation involving nanofluidic devices and improved emulsion chemistries has allowed for more widespread use of digital PCR, giving way to the current commercially available platforms[ 19 ]; of these, emulsion-based ddPCR has undergone huge growth in cancer research. In fact, a recent study has investigated the application of ddPCR to quantify mRNA biomarkers in stool from patients with CRC as a potential non-invasive screening test[ 17 ]. Thus, analysis of DNA obtained from fecal samples in patients with CRC may complement currently used procedures for diagnosis and disease follow-up.…”

Section: Introductionmentioning

confidence: 99%

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Detection of KRAS G12D in colorectal cancer stool by droplet digital PCR

Olmedillas‐López

Lévano-Linares

Alexandre

et al. 2017

WJG

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AIMTo assess KRAS G12D mutation detection by droplet digital PCR (ddPCR) in stool-derived DNA from colorectal cancer (CRC) patients.METHODSIn this study, tumor tissue and stool samples were collected from 70 patients with stage I-IV CRC diagnosed by preoperative biopsy. KRAS mutational status was determined by pyrosequencing analysis of DNA obtained from formalin-fixed paraffin-embedded (FFPE) tumor tissues. The KRAS G12D mutation was then analyzed by ddPCR in FFPE tumors and stool-derived DNA from patients with this point mutation. Wild-type (WT) tumors, as determined by pyrosequencing, were included as controls; analysis of FFPE tissue and stool-derived DNA by ddPCR was performed for these patients as well.RESULTSAmong the total 70 patients included, KRAS mutations were detected by pyrosequencing in 32 (45.71%), whereas 38 (54.29%) had WT tumors. The frequency of KRAS mutations was higher in left-sided tumors (11 located in the right colon, 15 in the left, and 6 in the rectum). The predominant point mutation was KRAS G12D (14.29%, n = 10), which was more frequent in early-stage tumors (I-IIA, n = 7). In agreement with pyrosequencing results, the KRAS G12D mutation was detected by ddPCR in FFPE tumor-derived DNA, and only a residual number of mutated copies was found in WT controls. The KRAS G12D mutation was also detected in stool-derived DNA in 80% of all fecal samples from CRC patients with this point mutation.CONCLUSIONddPCR is a reliable and sensitive method to analyze KRAS G12D mutation in stool-derived DNA from CRC patients, especially at early stages. This non-invasive approach is potentially applicable to other relevant biomarkers for CRC management.

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Droplet digital PCR for quantification ofITGA6in a stool mRNA assay for the detection of colorectal cancers

Cited by 16 publications

References 35 publications

Integrin α6β4 in Colorectal Cancer: Expression, Regulation, Functional Alterations and Use as a Biomarker

Integrin α6β4 in Colorectal Cancer: Expression, Regulation, Functional Alterations and Use as a Biomarker

Development of a reliable method to diagnose Streptococcus agalactiae infection by droplet digital PCR

Detection of KRAS G12D in colorectal cancer stool by droplet digital PCR

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