Assessment of Tumor Sequencing as a Replacement for Lynch Syndrome Screening and Current Molecular Tests for Patients With Colorectal Cancer.

Importance: Universal tumor screening for Lynch syndrome (LS) in colorectal cancer (CRC) is recommended and involves up to 6 sequential tests. Somatic gene testing is performed on stage IV CRCs for treatment determination. The diagnostic workup for patients with CRC could be simplified and improved using a single up-front tumor next-generation sequencing test if it has higher sensitivity and specificity than the current screening protocol. Objective: To determine whether up-front tumor sequencing (TS) could replace the current multiple sequential test approach for universal tumor screening for LS. Design, Setting, and Participants: Tumor DNA from 419 consecutive CRC cases undergoing standard universal tumor screening and germline genetic testing when indicated as part of the multicenter, population-based Ohio Colorectal Cancer Prevention Initiative from October 2015 through February 2016 (the prospective cohort) and 46 patients with CRC known to have LS due to a germline mutation in a mismatch repair gene from January 2013 through September 2015 (the validation cohort) underwent blinded TS. Main Outcomes and Measures: Sensitivity of TS compared with microsatellite instability (MSI) testing and immunohistochemical (IHC) staining for the detection of LS. Results: In the 465 patients, mean age at diagnosis was 59.9 years (range, 20-96 years), and 241 (51.8%) were female. Tumor sequencing identified all 46 known LS cases from the validation cohort and an additional 12 LS cases from the 419-member prospective cohort. Testing with MSI or IHC, followed by BRAF p.V600E testing missed 5 and 6 cases of LS, respectively. Tumor sequencing alone had better sensitivity (100%; 95% CI, 93.8%-100%) than IHC plus BRAF (89.7%; 95% CI, 78.8%-96.1%; P = .04) and MSI plus BRAF (91.4%; 95% CI, 81.0%-97.1%; P = .07). Tumor sequencing had equal specificity (95.3%; 95% CI, 92.6%-97.2%) to IHC plus BRAF (94.6%; 95% CI, 91.9%-96.6%; P > .99) and MSI plus BRAF (94.8%; 95% CI, 92.2%-96.8%; P = .88). Tumor sequencing identified 284 cases with KRAS, NRAS, or BRAF mutations that could affect therapy for stage IV CRC, avoiding another test. Finally, TS identified 8 patients with germline DPYD mutations that confer toxicity to fluorouracil chemotherapy, which could also be useful for treatment selection. Conclusions and Relevance: Up-front TS in CRC is simpler and has superior sensitivity to current multitest approaches to LS screening, while simultaneously providing critical information for treatment selection.

Challenges to "Classic" Esophageal Candidiasis: Looks Are Usually Deceiving.

Objectives: We undertook the first case control study of histologically confirmed esophageal candidiasis (EC). Methods: A computer search from July 2012 through February 2015 identified 1,011 esophageal specimens, including 40 cases of EC and 20 controls. Results: The EC incidence was 5.2%; it was associated with immunosuppression and endoscopic white plaques and breaks. Smoking was a predisposing factor, and alcohol was protective. EC had no unique symptoms, and 54% of endoscopic reports did not suspect EC. Important histologic clues included superficial and detached fragments of desquamated and hyper-pink parakeratosis, acute inflammation, intraepithelial lymphocytosis, dead keratinocytes, and bacterial overgrowth. Thirty percent had no neutrophilic infiltrate. Pseudohyphae were seen on H&E in 92.5% (n = 37/40). "Upfront" periodic acid-Schiff with diastase (PAS/D) on all esophageal specimens would have generated $68,333.49 in patient charges. Our targeted PAS/D strategy resulted in $13,044.87 in patient charges (cost saving = 80.9%, $55,288.62). Conclusions: We describe the typical morphology of EC and recommend limiting PAS/D to cases where the organisms are not readily identifiable on H&E and with at least one of the following: (1) ulcer, (2) suspicious morphology, and/or (3) clinical impression of EC.

A modified Lynch syndrome screening algorithm in colon cancer: BRAF immunohistochemistry is efficacious and cost beneficial.

OBJECTIVES: Somatic BRAF mutation in colon cancer essentially excludes Lynch syndrome. We compared BRAF V600E immunohistochemistry (IHC) with BRAF mutation in core, biopsy, and whole-section slides to determine whether IHC is similar and to assess the cost-benefit of IHC. METHODS: Resection cases (2009-2013) with absent MLH1 and PMS2 and prior BRAF mutation polymerase chain reaction results were chosen (n = 57). To mimic biopsy specimens, tissue microarrays (TMAs) were constructed. In addition, available biopsies performed prior to the resection were available in 15 cases. BRAF V600E IHC was performed and graded on TMAs, available biopsy specimens, and whole-section slides. Mutation status was compared with IHC, and cost-benefit analysis was performed. RESULTS: BRAF V600E IHC was similar in TMAs, biopsy specimens, and whole-section slides, with only four (7%) showing discordance between IHC and mutation status. Using BRAF V600E IHC in our Lynch syndrome screening algorithm, we found a 10% cost savings compared with mutational analysis. CONCLUSIONS: BRAF V600E IHC was concordant between TMAs, biopsy specimens, and whole-section slides, suggesting biopsy specimens are as useful as whole sections. IHC remained cost beneficial compared with mutational analysis, even though more patients needed additional molecular testing to exclude Lynch syndrome.

Comparative mutational profiling in the assessment of lung lesions: should it be the standard of care?

BACKGROUND: Discerning primary versus metastatic lung lesions is problematic. Comparative mutational profiling (CMP) involves genetic and point mutation analysis of lesions to facilitate this. We sought to review our experience in cases of two lung lesions or head and neck cancer and lung lesions to determine whether a significantly clinical problem existed, what standard processes were in place to address it, and whether a new diagnostic standard was required. METHODS: Between January 1, 2007, and October 31, 2008, CMP was used in 24 cases of two lung lesions or a head and neck cancer and lung lesion. Routine hematoxylin and eosin stain examination and immunohistochemistry were performed as appropriate. The CMP involved DNA sequencing for specific oncogene point mutations and a panel of allelic imbalance markers. Metastatic cancer required demonstration of concordant mutations affecting the same allele copy in different cancer deposits. RESULTS: The patient mean age was 62 years; there were 13 men and 11 women. The cases involved two lung lesions (n = 13) or a head and neck cancer and a lung lesion (n = 11). Standard pathology examination was unable to discriminate the lesions, and they were subsequently differentiated by CMP. Fifteen discordant CMP results were interpreted as independent primaries; 9 cases were concordant, consistent with metastatic disease. CONCLUSIONS: Discerning primary versus metastatic disease when dealing with lung lesions is a clinically significant problem. Comparative mutational profiling was found to be useful and reliable to assess the relatedness of multiple cancer lesions when routine pathology assessment was unable to.

To step or not to step: an approach to clinically diagnosed polyps with no initial pathologic finding.

We determined whether there were additional diagnostic findings in additional level sections performed on polyps with no pathologic diagnosis (NPD) or those in which only lymphoid aggregates (LAs) were seen initially and determined the level at which findings were identified. All colorectal biopsy specimens submitted with a clinical diagnosis of polyp during a 6-month period were included (N = 733). Initially, 3 level sections were cut for each polyp, and if a cause for the polyp was found, no additional levels were evaluated. If LAs or no cause for the polyp was found, 5 additional levels through each block were examined. Any diagnostic findings and the level at which they were identified were recorded. A discrete cause for the polyp was identified in routine levels in 574 cases (78.3%). Deeper levels were performed in 159: 23 for clarification of a suspected diagnosis, 38 for LAs, and 98 for NPD. Findings were identified in 31 (22.8%) of 136 stepped for LA or NPD with neoplastic findings in 13 (9.6%). Most diagnoses were identified in levels 4 or 5, but tubular adenomas were found in levels 7 and 8. These results support level sectioning specimens submitted as polyps with NPD or LAs on initial sections.