Diagnosis and risk stratification of Barrett’s dysplasia by flow cytometric DNA analysis of paraffin-embedded tissue

Choi, Won‐Tak; Tsai, Jia-Huei; Rabinovitch, Peter S.; Small, Thomas; Huang, Danning; Mattis, Aras N.; Kakar, Sanjay

doi:10.1136/gutjnl-2017-313815

Cited by 29 publications

(38 citation statements)

References 49 publications

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“…Of note, based on the published consensus guidelines, 20,21 the minimum number of cells or nuclei needed in a histogram to reproducibly detect aneuploidy is approximately 5000, which can be easily obtained from small mucosal biopsies. [14][15][16][17][18][19] The number of nuclei collected for DNA content measurements in this study ranged from 5000 to 20 000 (not including aggregates and debris), and the mean and median percentages of nuclei seen in an aneuploid population were 31 and 22% of this total, respectively (range = 6-70%).…”

Section: Materials and Methods P A T I E N T S A N D D A T A C O L L mentioning

confidence: 94%

“…As previously described, [14][15][16][17][18][19] three to six 60-lm-thick sections were cut from each tissue block, and the area of CCA was manually dissected for analysis. DNA fluorescent dye DAPI (4,6-diamidino-2phenylindole; Accurate Chemical & Scientific Corporation, Westbury, NY, USA) was used to stain each specimen, which was subsequently analysed on a BD LSRII S854 flow cytometer (BD Biosciences, San Jose, CA, USA) with UV laser excitation.…”

Section: Materials and Methods P A T I E N T S A N D D A T A C O L L mentioning

confidence: 99%

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DNA flow cytometric and interobserver study of crypt cell atypia in inflammatory bowel disease

Wen

Umetsu

Goldblum³

et al. 2019

Histopathology

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Aims The pathological features and diagnostic reliability of crypt cell atypia (CCA) arising in inflammatory bowel disease (IBD) and its clinical significance are unknown. Methods and results DNA flow cytometry (FCM) was performed on 14 colon biopsies of CCA from seven IBD patients (male‐to‐female ratio, 5:2; mean age, 53 years; mean IBD duration, 15 years) using paraffin‐embedded tissue. Seven gastrointestinal pathologists were asked to diagnose each biopsy as negative for dysplasia (NEG), indefinite for dysplasia (IND), low‐grade dysplasia (LGD) or high‐grade dysplasia (HGD) by morphology alone, then again with knowledge of FCM results. Aneuploidy was detected in all 14 biopsies, and five of eight biopsies (63%) also showed strong and diffuse nuclear staining for p53 in the areas of CCA. Six (86%) patients developed HGD (n = 5) or adenocarcinoma (n = 1) in the same colonic segment where CCA had been diagnosed within a mean follow‐up time of 27 months. No follow‐up information was available in the remaining one patient. When diagnoses were grouped as NEG or ‘atypical’ (including IND, LGD or HGD), the overall agreement rate of 76% (kappa = 0.51) based on morphology alone improved to 90% (kappa = 0.81) with knowledge of FCM results. Even when categorised as NEG or dysplasia (LGD or HGD) with each of the IND diagnoses reclassified into three categories (NEG, LGD or HGD) based on the degree of suspicion for dysplasia, the overall agreement rate of 63% (kappa = 0.25) based on morphology alone improved to 73% (kappa = 0.46) with knowledge of FCM results. However, when grouped as NEG, LGD or HGD, the overall agreement rate was less than 40% (kappa < 0.09) regardless of knowledge of FCM results. Conclusions The presence of aneuploidy, p53 positivity and development of HGD or adenocarcinoma on follow‐up indicate that CCA likely represents a dysplastic lesion (at least LGD) and is a histological marker of neoplastic progression. Although the grading of CCA, largely based on cytological abnormalities, is subject to significant interobserver variability, CCA can be histologically identified and should lead to a recommendation of increased endoscopic surveillance, especially if aneuploidy is detected.

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Section: Materials and Methods P A T I E N T S A N D D A T A C O L L mentioning

confidence: 94%

Section: Materials and Methods P A T I E N T S A N D D A T A C O L L mentioning

confidence: 99%

DNA flow cytometric and interobserver study of crypt cell atypia in inflammatory bowel disease

Wen

Umetsu

Goldblum³

et al. 2019

Histopathology

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“…As previously described [38,39,41], three to four 60micron thick sections were cut from each tissue block, and a potential abnormal cell population was enriched by manually dissecting the areas of dysplasia from the background normal tissue. DAPI (4,6-diamidino-2-phenylindole; Accurate Chemical & Scientific Corporation, Westbury, NY) was used to stain each sample, which was subsequently analyzed with a BD LSRII S854 flow cytometer (BD Biosciences, San Jose, CA) using UV laser excitation.…”

Section: Dna Flow Cytometrymentioning

confidence: 99%

“…As an alternative, DNA flow cytometry can potentially be useful as an adjunct method for the diagnosis and/or risk assessment of ampullary dysplasia. Although we recently demonstrated its utility in dysplasia of Barrett's esophagus [38], inflammatory bowel disease [39,40], and stomach [41], reports of such analyses on ampullary dysplasia are currently not available. Thus, this study examines the potential value of DNA content analysis in the diagnosis and/or risk stratification of ampullary dysplasia using formalin-fixed paraffin-embedded tissue obtained from endoscopic biopsies.…”

Section: Introductionmentioning

confidence: 99%

Diagnosis, risk stratification, and management of ampullary dysplasia by DNA flow cytometric analysis of paraffin-embedded tissue

et al. 2019

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The limited accuracy of endoscopic biopsy in detecting high-grade dysplasia or adenocarcinoma within ampullary adenoma or dysplasia has been reported. The natural history of ampullary dysplasia is also unclear, and there are no established guidelines to determine which patients with ampullary dysplasia require resection versus surveillance endoscopy. DNA flow cytometry was performed on 47 ampullary biopsies with low-grade dysplasia, 18 high-grade dysplasia, and 23 negative for dysplasia, as well as 11 cases of ampullary adenocarcinoma. Abnormal DNA content (aneuploidy or elevated 4N fraction > 6%) was identified in 9 (82%) of adenocarcinoma, 13 (72%) of high-grade dysplasia, 7 (15%) of low-grade dysplasia, and none (0%) of non-dysplastic mucosa. One-, 2-, and 7-year detection rates of high-grade dysplasia or adenocarcinoma in low-grade dysplasia patients with abnormal DNA content were 57%, 86%, and 88%, respectively, whereas low-grade dysplasia patients in the setting of normal DNA content had 1-, 2-, and 7-year detection rates of 10%, 10%, and 10%, respectively. The univariate and multivariate hazard ratios (HRs) for subsequent detection of high-grade dysplasia or adenocarcinoma in low-grade dysplasia patients with DNA content abnormality were 16.8 (p = <0.01) and 9.8 (p = <0.01), respectively. Among the 13 high-grade dysplasia patients with DNA content abnormality, 5 patients (38%) were subsequently found to have adenocarcinoma within a mean follow-up time of 3 months, whereas only 1 (20%) of the remaining 5 patients in the setting of normal DNA content developed adenocarcinoma in a month (HR = 2.6, p = 0.39). The overall 1-and 2-year detection rates of adenocarcinoma in all high-grade dysplasia patients (regardless of flow cytometric results) were 34% (95% confidence interval = 16-63%) and 47% (95% confidence interval = 23-79%), respectively. In conclusion, the majority of low-grade dysplasia patients (86%) in the setting of abnormal DNA content developed high-grade dysplasia or adenocarcinoma within 2 years and thus may benefit from resection, whereas those with normal DNA content may be followed with surveillance endoscopy. The presence of DNA content abnormality can also confirm a morphologic suspicion of high-grade dysplasia, which should be managed with resection, as nearly 50% of the high-grade dysplasia patients were found to have adenocarcinoma within 2 years.

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“…18q loss was found in 32% BE, 42% LGD, 73% HGD, and 69% EAC [74]. One recent study has demonstrated the potential clinical application of DNA flow cytometry in the diagnosis and risk stratification for BE patients [75].…”

Section: Genetic and Molecular Signature Of Be And Early Stage Eacmentioning

confidence: 99%

Histopathology of Barrett’s Esophagus and Early-Stage Esophageal Adenocarcinoma: An Updated Review

Yin

Gonzálo

Lai

et al. 2018

Gastrointestinal Disorders

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Esophageal adenocarcinoma carries a very poor prognosis. For this reason, it is critical to have cost-effective surveillance and prevention strategies and early and accurate diagnosis, as well as evidence-based treatment guidelines. Barrett’s esophagus is the most important precursor lesion for esophageal adenocarcinoma, which follows a defined metaplasia–dysplasia–carcinoma sequence. Accurate recognition of dysplasia in Barrett’s esophagus is crucial due to its pivotal prognostic value. For early-stage esophageal adenocarcinoma, depth of submucosal invasion is a key prognostic factor. Our systematic review of all published data demonstrates a “rule of doubling” for the frequency of lymph node metastases: tumor invasion into each progressively deeper third of submucosal layer corresponds with a twofold increase in the risk of nodal metastases (9.9% in the superficial third of submucosa (sm1) group, 22.0% in the middle third of submucosa (sm2) group, and 40.7% in deep third of submucosa (sm3) group). Other important risk factors include lymphovascular invasion, tumor differentiation, and the recently reported tumor budding. In this review, we provide a concise update on the histopathological features, ancillary studies, molecular signatures, and surveillance/management guidelines along the natural history from Barrett’s esophagus to early stage invasive adenocarcinoma for practicing pathologists.

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Diagnosis and risk stratification of Barrett’s dysplasia by flow cytometric DNA analysis of paraffin-embedded tissue

Cited by 29 publications

References 49 publications

DNA flow cytometric and interobserver study of crypt cell atypia in inflammatory bowel disease

DNA flow cytometric and interobserver study of crypt cell atypia in inflammatory bowel disease

Diagnosis, risk stratification, and management of ampullary dysplasia by DNA flow cytometric analysis of paraffin-embedded tissue

Histopathology of Barrett’s Esophagus and Early-Stage Esophageal Adenocarcinoma: An Updated Review

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