Therapeutic and Prognostic Implications of BRAF V600E in Pediatric Low-Grade Gliomas

Purpose BRAF V600E is a potentially highly targetable mutation detected in a subset of pediatric low-grade gliomas (PLGGs). Its biologic and clinical effect within this diverse group of tumors remains unknown. Patients and Methods A combined clinical and genetic institutional study of patients with PLGGs with long-term follow-up was performed (N = 510). Clinical and treatment data of patients with BRAF V600E mutated PLGG (n = 99) were compared with a large international independent cohort of patients with BRAF V600E mutated-PLGG (n = 180). Results BRAF V600E mutation was detected in 69 of 405 patients (17%) with PLGG across a broad spectrum of histologies and sites, including midline locations, which are not often routinely biopsied in clinical practice. Patients with BRAF V600E PLGG exhibited poor outcomes after chemotherapy and radiation therapies that resulted in a 10-year progression-free survival of 27% (95% CI, 12.1% to 41.9%) and 60.2% (95% CI, 53.3% to 67.1%) for BRAF V600E and wild-type PLGG, respectively (P < .001). Additional multivariable clinical and molecular stratification revealed that the extent of resection and CDKN2A deletion contributed independently to poor outcome in BRAF V600E PLGG. A similar independent role for CDKN2A and resection on outcome were observed in the independent cohort. Quantitative imaging analysis revealed progressive disease and a lack of response to conventional chemotherapy in most patients with BRAF V600E PLGG. Conclusion BRAF V600E PLGG constitutes a distinct entity with poor prognosis when treated with current adjuvant therapy. (C) 2017 by American Society of Clinical Oncolog

Therapeutic and Prognostic Implications of BRAF V600E in Pediatric Low-Grade Gliomas.

Purpose BRAF V600E is a potentially highly targetable mutation detected in a subset of pediatric low-grade gliomas (PLGGs). Its biologic and clinical effect within this diverse group of tumors remains unknown. Patients and Methods A combined clinical and genetic institutional study of patients with PLGGs with long-term follow-up was performed (N = 510). Clinical and treatment data of patients with BRAF V600E mutated PLGG (n = 99) were compared with a large international independent cohort of patients with BRAF V600E mutated-PLGG (n = 180). Results BRAF V600E mutation was detected in 69 of 405 patients (17%) with PLGG across a broad spectrum of histologies and sites, including midline locations, which are not often routinely biopsied in clinical practice. Patients with BRAF V600E PLGG exhibited poor outcomes after chemotherapy and radiation therapies that resulted in a 10-year progression-free survival of 27% (95% CI, 12.1% to 41.9%) and 60.2% (95% CI, 53.3% to 67.1%) for BRAF V600E and wild-type PLGG, respectively ( P \u3c .001). Additional multivariable clinical and molecular stratification revealed that the extent of resection and CDKN2A deletion contributed independently to poor outcome in BRAF V600E PLGG. A similar independent role for CDKN2A and resection on outcome were observed in the independent cohort. Quantitative imaging analysis revealed progressive disease and a lack of response to conventional chemotherapy in most patients with BRAF V600E PLGG. Conclusion BRAF V600E PLGG constitutes a distinct entity with poor prognosis when treated with current adjuvant therapy

PD-L1 immunostaining scoring for non-small cell lung cancer based on immunosurveillance parameters - Table 2

<p>PD-L1 immunostaining scoring for non-small cell lung cancer based on immunosurveillance parameters</p> - Table

Repeated Traumatic Brain Injury Affects Composite Cognitive Function in Piglets

Cumulative effects of repetitive mild head injury in the pediatric population are unknown. We have developed a cognitive composite dysfunction score that correlates white matter injury severity in neonatal piglets with neurobehavioral assessments of executive function, memory, learning, and problem solving. Anesthetized 3- to 5-day-old piglets were subjected to single (n = 7), double one day apart (n = 7), and double one week apart (n = 7) moderate (190 rad/s) rapid non-impact axial rotations of the head and compared to instrumented shams (n = 7). Animals experiencing two head rotations one day apart had a significantly higher mortality rate (43%) compared to the other groups and had higher failures rates in visual-based problem solving compared to instrumented shams. White matter injury, assessed by β-APP staining, was significantly higher in the double one week apart group compared to that with single injury and sham. Worsening performance on cognitive composite score correlated well with increasing severity of white matter axonal injury. In our immature large animal model of TBI, two head rotations produced poorer outcome as assessed by neuropathology and neurobehavioral functional outcomes compared to that with single rotations. More importantly, we have observed an increase in injury severity and mortality when the head rotations occur 24 h apart compared to 7 days apart. These observations have important clinical translation to infants subjected to repeated inflicted head trauma

Classification of NSCLC according to PD-L1 expression patterns in tumor-nest and TME compartments.

<p>Classification of NSCLC according to PD-L1 expression patterns in tumor-nest and TME compartments.</p

Relationships between biomarkers in our NSCLC cohort.

<p><b>(A)</b> Hierarchical clustering heat map of percentages of positive cells for FOXP3, PD-L1, CD8, Granzyme B, and CD68. Each biomarker (row) was normalized before clustering to give a mean of 0 and a standard deviation of 1 across columns (samples). The color scale indicates the relative size of the biomarker score compared to the other samples in the cohort with blue = low, white = neutral, and brown = high. Bracket height indicates the degree of correlation between measurements with shorter brackets representing higher correlations and taller brackets representing smaller correlations. <b>(B)</b> Heatmap of Pearson correlation coefficients between percent positive cells measurements across samples for FOXP3, PD-L1, CD8, Granzyme B, and CD68. The color scale ranges from the minimum (blue) and maximum (red) correlation coefficient observed in this analysis. Therefore red represents strong positive correlations, white represents moderate correlations, and blue represents weak correlations. No strong negative correlations were observed.</p

Examples of PD-L1 staining patterns observed and categorized in this NSCLC series.

<p><b>A</b>: Pattern I—Constitutive mixed with induced. Diffuse expression of PD-L1 (IHC) on tumor cell membranes of a squamous cell carcinoma, including central regions of trabeculae. Prominent labeling of cells in the TME compartment at the tumor-nest-TME interface suggesting presence of an immunological synapse (inset arrow). <b>B</b>: Pattern II—Induced only. Patchy expression of PD-L1 in a squamous cell carcinoma at the tumor-nest-TME interface (inset arrow). Minimal to no PD-L1 expression in the trabeculae (asterisk) if compared with (<b>A</b>). <b>C</b>: Pattern III—Immune ignorance. No to minimal PD-L1 expression in both tumor and TME compartments in an adenocarcinoma. <b>D</b>: Pattern IV—Constitutive only. Diffuse expression of PD-L1 by tumor-nests in an adenocarcinoma with minimal TME staining. <b>E</b>: Pattern V—TME expression only. No to minimal PD-L1 expression in tumor cells of a squamous cell carcinoma, with widespread staining in the TME compartment. <b>F</b>: Percentages of PD-L1-positive tumor cells in all staining pattern categories (I-V). <b>G</b>: Percentages of TME PD-L1-positive cells in all staining pattern categories (I-V). <b>H</b>: Digital H-scores from tumor-nests in all staining pattern categories (I-V). Significant differences between groups I and II were calculated using two-tailed Student’s t-tests, assuming unequal sample variances (*<i>P</i> ≤ 0.05, **<i>P</i> ≤ 0.01, ***<i>P</i> ≤ 0.001 and ****<i>P</i> ≤ 0.0001). Scale bars: 200 μm (<b>A-E</b>). NSCLC = non-small cell lung carcinoma; IHC = immunohistochemistry; TME = tumor microenvironment.</p

Example of computational tissue analysis (cTA) of NSCLC sections.

<p><b>A</b>: Manual inclusion (green line) and exclusion (red line) annotations in squamous cell carcinoma tissue for CD56 IHC. <b>B</b>: Enlarged view area from (<b>A)</b> showing exclusion annotation around a central necrotic area in tumor-nest (arrow) and a segment of the inclusion annotation around the tumor mass margin (arrowhead). <b>C</b>: PD-L1 IHC of a squamous cell carcinoma. <b>D</b>: Algorithm markup from (<b>C)</b> showing membrane scoring only in cells assigned to the tumor-nest compartment, with 1+ = yellow, 2+ = orange, and 3+ = red. The separated TME compartment cell nuclei are marked in green. Scale bars: 1 mm (<b>A</b>); 100 μm (<b>B</b>); and 150 μm (<b>C</b> and <b>D</b>). NSCLC = non-small cell lung carcinoma; IHC = immunohistochemistry; TME = tumor microenvironment.</p