Correlation of tumor microenvironment from biopsy and resection specimens in untreated colorectal cancer patients: a surprising lack of agreement.

BACKGROUND: Colorectal cancer (CRC) tumor microenvironment (TME) characteristics, such as tumor infiltrating lymphocyte (TIL) densities and PD-L1 status, are predictive of recurrence, disease-free survival, and overall survival. In many malignancies, TME characteristics are also predictive of response to immunotherapy. As window of opportunity studies using neoadjuvant immunotherapy become more common and treatment guidelines incorporate TME features, accurate assessment of the pre-treatment TME using the biopsy specimen is critical. However, no study has thoroughly evaluated the correlation between the TMEs of the biopsy and resection specimens. METHODS: We conducted a retrospective analysis of patients with stage I-III CRC with matched biopsy and resection specimens. CD3+, CD4+, CD8+, and FoxP3+ lymphocyte populations at the center of tumor (CT) and invasive margin (IM) and tumor PD-L1 status in the biopsy and resection specimens were evaluated. TIL populations were compared using Mann-Whitney U tests or Student's t tests and correlated using Pearson r. RESULTS: CD3+ and CD4+ densities were significantly higher in the CT of the biopsy relative to the resection specimen Comparing biopsy and resection specimens, no TIL population at either the CT or IM had a correlation coefficient > 0.5. Determining PD-L1 status based on biopsy tissue resulted in a sensitivity of 37.1%, specificity of 81.4%, and accuracy of 61.5%. CONCLUSIONS: These findings demonstrate significant discordance between the TME of the biopsy and resection specimens. Caution should be used when basing treatment decisions on pre-treatment endoscopic biopsy findings and when interpreting changes in the TME between pre-treatment biopsy and resection specimens after neoadjuvant therapy.

Development and Validation of Ultra-Rapid Periodic Acid-Schiff Stain for Use in Identifying Fungus on Frozen Section.

CONTEXT.­: There is no standardized process for utilization of periodic acid-Schiff during intraoperative frozen sections to identify fungal organisms. OBJECTIVE.­: To develop a rapid staining process for fresh tissue with periodic acid-Schiff during intraoperative consultation and develop an appropriate control block. DESIGN.­: Muscle tissue was inoculated with 2 species of fungus (Aspergillus fumigatus and Paecilomyces spp) and grown at 3 different temperatures for 72 hours. Inoculated tissue was embedded in optimal cutting temperature compound, cut, and stained using a modified periodic acid-Schiff stain. The optimal control was determined for future use as the standard control. Multiple control slides were cut and stained, using successively shorter time intervals for each step. The staining process that provided accurate results in the shortest amount of time was deemed ultra-rapid periodic acid-Schiff. This method was validated by carryover studies and clinical specimens. RESULTS.­: Paecilomyces spp incubated at 30°C for 72 hours was the most optimal positive control, with numerous yeast and hyphal forms. The fastest staining process involved 2 minutes of periodic acid and Schiff reagent and 10 dips of light green solution. Tap water was as effective as distilled water. Validation was successfully achieved. Clinical cases all stained identical to formalin-fixed, paraffin-embedded tissue stained with hematoxylin-eosin and periodic acid-Schiff. CONCLUSIONS.­: Ultra-rapid periodic acid-Schiff provides fast and reliable identification of fungal organisms on fresh tissue. Development of a concurrent positive control allows for quality control and validation.