Innate immune activation by checkpoint inhibition in human patient-derived lung cancer tissues.

Although Pembrolizumab-based immunotherapy has significantly improved lung cancer patient survival, many patients show variable efficacy and resistance development. A better understanding of the drug's action is needed to improve patient outcomes. Functional heterogeneity of the tumor microenvironment (TME) is crucial to modulating drug resistance; understanding of individual patients' TME that impacts drug response is hampered by lack of appropriate models. Lung organotypic tissue slice cultures (OTC) with patients' native TME procured from primary and brain-metastasized (BM) non-small cell lung cancer (NSCLC) patients were treated with Pembrolizumab and/or beta-glucan (WGP, an innate immune activator). Metabolic tracing with 13C6-Glc/13C5,15N2-Gln, multiplex immunofluorescence, and digital spatial profiling (DSP) were employed to interrogate metabolic and functional responses to Pembrolizumab and/or WGP. Primary and BM PD-1+ lung cancer OTC responded to Pembrolizumab and Pembrolizumab + WGP treatments, respectively. Pembrolizumab activated innate immune metabolism and functions in primary OTC, which were accompanied by tissue damage. DSP analysis indicated an overall decrease in immunosuppressive macrophages and T cells but revealed microheterogeneity in immune responses and tissue damage. Two TMEs with altered cancer cell properties showed resistance. Pembrolizumab or WGP alone had negligible effects on BM-lung cancer OTC but Pembrolizumab + WGP blocked central metabolism with increased pro-inflammatory effector release and tissue damage. In-depth metabolic analysis and multiplex TME imaging of lung cancer OTC demonstrated overall innate immune activation by Pembrolizumab but heterogeneous responses in the native TME of a patient with primary NSCLC. Metabolic and functional analysis also revealed synergistic action of Pembrolizumab and WGP in OTC of metastatic NSCLC.

Predictive value of phenotypic signatures of bladder cancer response to cisplatin-based neoadjuvant chemotherapy.

BACKGROUND: Cisplatin-based neoadjuvant chemotherapy (NAC) for muscle-invasive urothelial carcinoma of the bladder confers only a modest survival advantage. Patients with pathologic progression on NAC have poor outcomes related to a delay in definitive surgical management. OBJECTIVE: To characterize the value of epithelial-mesenchymal transition (EMT) effectors and other novel biomarkers to predict response to NAC. METHODS: A tissue microarray was constructed from patients with clinical stage T2 urothelial carcinoma of the bladder using transurethral resection (TUR) specimens (N = 69) and case-matched post-NAC cystectomy specimens (N = 51). Patients were stratified based on pathologic response to NAC and cancer-specific survival. Biomarker expression in TUR specimens was correlated with pathologic response to NAC and clinical outcomes. Phenotypic changes in expression induced by cisplatin-based NAC were characterized in primary TUR and post-NAC cystectomy and lymph node metastasis specimens. RESULTS: Increased expression of mesenchymal markers and actin-cytoskeleton regulators, as well as low apoptosis index, in TUR specimens was associated with pathologic progression on cisplatin-based NAC. Overexpression of N-cadherin and decreased apoptosis was predictive of disease-specific mortality. NAC decreased cofilin phosphorylation and induced a mesenchymal-epithelial transition phenotype. CONCLUSIONS: The epithelial-mesenchymal transition phenotype and actin-cytoskeleton remodeling are associated with pathologic progression on NAC. Chemotherapy induced an mesenchymal-epithelial transition phenotype and decreased cofilin phosphorylation, providing new insights into therapeutic resistance mechanisms. Primary tumors with high apoptosis rates exhibited favorable response to NAC and lower mortality rates, indicating that these tumors may be sensitized to therapy. Further validation will establish these novel signatures as predictors of therapeutic response.

Prostate cancer polar localization on core biopsy predicts pathologic stage.

INTRODUCTION: This study investigated the polar sub-localization of prostate cancer on needle core biopsy ('polar' defined as tumor = 1 mm from the tissue polar edge) as a predictor of extraprostatic extension. MATERIALS AND METHODS: Histologic sections from 58 patients who underwent preoperative prostate biopsy and radical prostatectomy at the University of Kentucky from 2006 to 2013 were evaluated. Patients were retrospectively case matched based on pathologic stage (pT2 versus pT3/4) using biopsy Gleason grade and prostate-specific antigen. Histologic sections of needle core biopsies were analyzed for polar involvement. The location of polar involvement was correlated to the presence of extraprostatic extension on final prostatectomy pathology. RESULTS: Average percentage of total polar cores was predictive of extraprostatic extension on final prostatectomy, particularly in the prostatic apex and base (p = 0.029 and 0.006, respectively). Higher grade tumors were identified at the pole in the high stage cohort (p = 0.032). Total percent polar involvement had the greatest sensitivity and specificity for predicting extraprostatic extension when directly compared to previously described histologic parameters (percent greatest involvement of a single core, length of greatest involvement of a single core, presence of perineural invasion, presence of bilateral gland involvement, and percent total positive core involvement). The location of polar involvement on needle core biopsy was also predictive of the precise location of extraprostatic extension on final prostatectomy pathology (Chi-square p < .001, negative predictive value > 70% in all prostate sextants). CONCLUSIONS: These data suggest the use of biopsy polar core involvement as a valuable histologic predictor of increased pathologic stage.

Differential effects of post-implantation time on potassium- versus D-amphetamine-evoked dopamine overflow in the striatum of F344 rats.

Effects of post-implantation time on potassium (K+)- versus D-amphetamine (D-AMPH)-evoked striatal dopamine (DA) overflow were measured using microdialysis in freely moving young and aged Fischer 344 rats. In one group, samples were collected on the day of probe insertion (Day 1 group). In a second group, samples were collected 24 h after probe insertion (Day 2 group). While analyses revealed no significant differences between the two age groups, the 100 mM K+ stimulus evoked a significantly greater amount of DA overflow in the Day 1 group compared to the Day 2 group. The decrease in 3,4-dihydroxyphenylacetic acid (DOPAC) produced by K+ stimulation was not influenced by post-implantation time. The effect of the 250 microM D-AMPH stimulus on DA overflow did not differ between the Day 1 and Day 2 groups, nor did the decrease in DOPAC that accompanied D-AMPH stimulation. These results support the hypothesis that under some stimulus conditions, post-implantation time is an important variable in microdialysis studies.

Microdialysis studies of D-amphetamine-evoked striatal dopamine overflow in young versus aged F344 rats: effects of concentration and order of administration.

In order to measure the effects of different concentrations of D-amphetamine (D-AMPH) infusions on striatal dopamine (DA) overflow in young versus aged rats, and to determine the influence of preceding infusions on subsequent stimuli, two microdialysis studies were conducted. In the first study, D-AMPH (100, 200, and 2000 microM) was infused in ascending order of concentration, while in the second study the order of administration was reversed. The order of administration significantly affected DA overflow and extracellular levels of 3,4-dihydroxyphenylacetic acid (DOPAC). Generally, DA overflow was greater for a given concentration when it was administered first in a sequence than when it was administered later in a sequence. The two age groups did not differ on measures of DA overflow. The order of administration also significantly influenced the effect of D-AMPH on extracellular DOPAC, as the D-AMPH-related decreases in DOPAC were greater for a concentration when it was administered earlier versus later in the sequence. This effect was greater in the young rats than in the aged rats. D-AMPH also resulted in diminished levels of DOPAC in the aged rats compared to the young rats. These results suggest that between-groups studies may be more appropriate for determining the effects of different concentrations of D-AMPH on striatal DA overflow. They also demonstrate that while some measures of stimulus-evoked DA overflow may not differ between young and aged F344 rats, extracellular regulation of striatal DA (as measured by changes in DOPAC) may be altered. These alterations may contribute to age-related decreases in motor function.