Differential regulation of PD-L1 expression by immune and tumor cells in NSCLC and the response to treatment with atezolizumab (anti-PD-L1).

Programmed death-ligand 1 (PD-L1) expression on tumor cells (TCs) by immunohistochemistry is rapidly gaining importance as a diagnostic for the selection or stratification of patients with non-small cell lung cancer (NSCLC) most likely to respond to single-agent checkpoint inhibitors. However, at least two distinct patterns of PD-L1 expression have been observed with potential biological and clinical relevance in NSCLC: expression on TC or on tumor-infiltrating immune cells (ICs). We investigated the molecular and cellular characteristics associated with PD-L1 expression in these distinct cell compartments in 4,549 cases of NSCLC. PD-L1 expression on IC was more prevalent and likely reflected IFN-γ-induced adaptive regulation accompanied by increased tumor-infiltrating lymphocytes and effector T cells. High PD-L1 expression on TC, however, reflected an epigenetic dysregulation of the PD-L1 gene and was associated with a distinct histology described by poor immune infiltration, sclerotic/desmoplastic stroma, and mesenchymal molecular features. Importantly, durable clinical responses to atezolizumab (anti-PD-L1) were observed in patients with tumors expressing high PD-L1 levels on either TC alone [40% objective response rate (ORR)] or IC alone (22% ORR). Thus, PD-L1 expression on TC or IC can independently attenuate anticancer immunity and emphasizes the functional importance of IC in regulating the antitumor T cell response.

A Process Evaluation of the Skin Cancer Prevention Act (Tanning Beds): A Survey of Ontario Public Health Units.

Evidence of the dangers of indoor tanning and its popularity, including among youth, led the Government of Ontario to pass the Skin Cancer Prevention Act (Tanning Beds) (SCPA) in 2014. This legislation includes prohibiting the sale of indoor tanning services to individuals under 18, requiring warning signs be posted, and other safety regulations. We collected information from Ontario Public Health Units to conduct a process evaluation of the SCPA to: understand legislation implementation; assess available evidence about compliance, inspection, and enforcement; and, note barriers and facilitators related to inspection and enforcement. Data was collected March-April 2018. All 36 Ontario Public Health Units were invited to participate in an online questionnaire about the SCPA. Questions covered complaints, inspection, and enforcement, and used both close- and open-ended questions. Participants from 20 Public Health Units responded to the questionnaire; a response rate of 56%. These agencies reported 485 facilities offer indoor tanning. Since 2014, there have been 242 infractions by tanning facility owner/operators related to the SCPA, with most being uncovered during non-mandatory routine inspections (n = 234, 97%), rather than mandatory complaint-based inspections (n = 8, 3%). Most infractions were related to warning signs (n = 201, 83%). No charges were issued for any infractions. Instead, providing education (n = 90, 62%) and issuing warnings (n = 33, 23%) were the most common enforcement strategies. SCPA amendments are needed, including mandatory, routinely scheduled inspections. In addition to providing education, fines may improve compliance. More resources are required for inspection and enforcement of the SCPA.

Quantitative determination of cellular farnesyltransferase activity: towards defining the minimum substrate reactivity for biologically relevant protein farnesylation.

Prenylation is a post-translational modification wherein an isoprenoid group is attached to a protein substrate by a protein prenyltransferase. Hundreds of peptide sequences are in vitro substrates for protein farnesyltransferase (FTase), but it remains unknown which of these sequences can successfully compete for in vivo prenylation. Translating in vitro studies to predict in vivo protein farnesylation requires determining the minimum reactivity needed for modification by FTase within the cell. Towards this goal, we developed a reporter protein series spanning several orders of magnitude in FTase reactivity as a calibrated sensor for endogenous FTase activity. Our approach provides a minimally invasive method to monitor changes in cellular FTase activity in response to environmental or genetic factors. Determining the reactivity "threshold" for in vivo prenylation will help define the prenylated proteome and identify prenylation-dependent pathways for therapeutic targeting.

C-reactive protein reduction post treatment is associated with improved survival in atezolizumab (anti-PD-L1) treated non-small cell lung cancer patients.

PURPOSE: Overall survival (OS) is the most significant endpoint for evaluation of treatment benefit with checkpoint inhibitors (CPI) in cancer. We evaluated serum C-reactive protein (CRP) in non-small cell lung cancer (NSCLC) trials with atezolizumab (anti-PD-L1) as an early OS surrogate. METHODS: Serum from patients enrolled in randomized Phase II (n = 240) and Phase III (n = 701) trials of NSCLC patients (POPLAR, OAK) who progressed on prior-platinum chemotherapy, were analyzed for CRP levels over time. Patients were grouped by changes in CRP levels post-treatment as either increased (≥ 1.5 fold), decreased (≤ 1.5 fold) or unchanged (within +1.5 fold) relative to pre-treatment levels to assess association with progression free survival (PFS) and OS. RESULTS: Decrease in serum CRP levels at 6 weeks relative to pre-treatment were observed in patients with RECIST1.1 based complete or partial responses (CR/PR) to atezolizumab whereas patients with disease progression (PD) demonstrated an increase in CRP levels in the Phase II POPLAR study, and confirmed in the Phase III OAK study. Decrease in serum CRP as early as six weeks post treatment predicted improved PFS and OS, even in patients who were determined as stable disease (SD) in their first scan. This effect was not observed in the chemotherapy arms. CONCLUSION: Modulation of serum CRP correlates with clinical outcome post-atezolizumab treatment. This routine lab test may provide utility in informing OS signals as early as 6 weeks post-initiation of therapy with CPIs in NSCLC.

Survey design from the ground up: collaboratively creating the Toronto Teen Survey.

The Toronto Teen Survey is a community-based participatory research study whose aim is to gather information on the accessibility and relevance of sexual health services for diverse groups of urban youth (13 to 17 years of age). This information will be used to develop a proactive, citywide strategy to improve sexual health outcomes for Toronto adolescents. In this article, the authors focus on the processes of collaboratively developing a survey tool with youth, academics, and community stakeholders. An overview of the project and examples from the design stage are provided. In addition, recommendations are given toward developing best practices when working with young people on research and survey design.

Late-differentiated effector neoantigen-specific CD8+ T cells are enriched in peripheral blood of non-small cell lung carcinoma patients responding to atezolizumab treatment.

BACKGROUND: There is strong evidence that immunotherapy-mediated tumor rejection can be driven by tumor-specific CD8+ T cells reinvigorated to recognize neoantigens derived from tumor somatic mutations. Thus, the frequencies or characteristics of tumor-reactive, mutation-specific CD8+ T cells could be used as biomarkers of an anti-tumor response. However, such neoantigen-specific T cells are difficult to reliably identify due to their low frequency in peripheral blood and wide range of potential epitope specificities. METHODS: Peripheral blood mononuclear cells (PBMC) from 14 non-small cell lung cancer (NSCLC) patients were collected pre- and post-treatment with the anti-PD-L1 antibody atezolizumab. Using whole exome sequencing and RNA sequencing we identified tumor neoantigens that are predicted to bind to major histocompatibility complex class I (MHC-I) and utilized mass cytometry, together with cellular 'barcoding', to profile immune cells from patients with objective response to therapy (n = 8) and those with progressive disease (n = 6). In parallel, a highly-multiplexed combinatorial tetramer staining was used to screen antigen-specific CD8+ T cells in peripheral blood for 782 candidate tumor neoantigens and 71 known viral-derived control peptide epitopes across all patient samples. RESULTS: No significant treatment- or response associated phenotypic difference were measured in bulk CD8+ T cells. Multiplexed peptide-MHC multimer staining detected 20 different neoantigen-specific T cell populations, as well as T cells specific for viral control antigens. Not only were neoantigen-specific T cells more frequently detected in responding patients, their phenotypes were also almost entirely distinct. Neoantigen-specific T cells from responder patients typically showed a differentiated effector phenotype, most like Cytomegalovirus (CMV) and some types of Epstein-Barr virus (EBV)-specific CD8+ T cells. In contrast, more memory-like phenotypic profiles were observed for neoantigen-specific CD8+ T cells from patients with progressive disease. CONCLUSION: This study demonstrates that neoantigen-specific T cells can be detected in peripheral blood in non-small cell lung cancer (NSCLC) patients during anti-PD-L1 therapy. Patients with an objective response had an enrichment of neoantigen-reactive T cells and these cells showed a phenotype that differed from patients without a response. These findings suggest the ex vivo identification, characterization, and longitudinal follow-up of rare tumor-specific differentiated effector neoantigen-specific T cells may be useful in predicting response to checkpoint blockade. TRIAL REGISTRATION: POPLAR trial NCT01903993 .

Predicting which childhood memories persist: contributions of memory characteristics.

This investigation identified memory-level predictors of the survivability of 4- to 13-year-old children's earliest recollections over a 2-year period. Data previously reported by Peterson, Warren, and Short (2011) were coded for inclusion of emotion terms and thematic, chronological, and contextual narrative coherence. In addition, the uniqueness and content of the reported events were classified, and the presence or absence of event reminders was recorded. The use of logistic multilevel modeling indicated that emotion and each dimension of coherence added to the prediction of a memory's survivability over and above age-related variance. In contrast, event uniqueness, content category, reminders, and word count were not associated with retention. The findings help explain why particular early memories endure over time.

Factors affecting mortality in a large cohort study with special reference to oral contraceptive use.

BACKGROUND: This analysis updates mortality in the Oxford-Family Planning Association (Oxford-FPA) contraceptive study, with emphasis on oral contraceptive (OC) use. STUDY DESIGN: The Oxford-FPA study includes 17,032 women recruited from 1968-1974 at contraceptive clinics, aged 25-39 years, using OCs a diaphragm or an intrauterine device. Follow-up has been to March 2009; by then, 1715 women had died. RESULTS: The rate ratio (RR) for overall mortality was 0.87 (CI 0.79-0.96), comparing ever-users of OCs with never-users. The RR for fatal cervical cancer was increased (7.3), but the CIs were very wide (1.2-305). There was no association between ever-use of OCs and mortality from breast cancer (RR 1.0, CI 0.8-1.2), nor was fatal breast cancer related to duration of OC use. OC use strongly protected against death from other uterine cancer and ovarian cancer; RRs for ever-use of OCs were 0.3 (CI 0.1-0.8) and 0.4 (CI 0.3-0.6), respectively. Protection increased with duration of OC use and persisted more than 20 years after cessation. Circulatory disease mortality was not increased, the RR for ever-use of OCs being 0.9 (CI 0.7-1.1). The overall mortality RR for all women smoking 15+ cigarettes daily was 2.25 (CI 1.99-2.53) and, for all women with a body mass index of 28+ kg/m(2), was 1.33 (CI 1.07-1.64). CONCLUSIONS: Long-term follow-up strongly suggests that OC use slightly reduces all cause mortality.