Molecular imaging of drug transit through the blood-brain barrier with MALDI mass spectrometry imaging

Drug transit through the blood-brain barrier (BBB) is essential for therapeutic responses in malignant glioma. Conventional methods for assessment of BBB penetrance require synthesis of isotopically labeled drug derivatives. Here, we report a new methodology using matrix assisted laser desorption ionization mass spectrometry imaging (MALDI MSI) to visualize drug penetration in brain tissue without molecular labeling. In studies summarized here, we first validate heme as a simple and robust MALDI MSI marker for the lumen of blood vessels in the brain. We go on to provide three examples of how MALDI MSI can provide chemical and biological insights into BBB penetrance and metabolism of small molecule signal transduction inhibitors in the brain – insights that would be difficult or impossible to extract by use of radiolabeled compounds

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

The evolution of melanoma resistance reveals therapeutic opportunities

The RAS-RAF-MEK-ERK pathway is a key driver of proliferation and survival signals in tumor cells and has been the focus of intense drug development efforts over the past 20 years. The recent regulatory approval of RAF inhibitors and a MAP-ERK kinase (MEK) inhibitor for metastatic melanoma provides clinical validation of tumor dependency on this pathway. Unfortunately, the therapeutic benefit of these agents is often short lived and resistance develops within a matter of months. Preclinical models of resistance to vemurafenib have provided critical insights into predicting, validating, and characterizing potential mechanisms. A key observation has been that vemurafenib-resistant tumor cells suffer a fitness deficit in the absence of drug treatment and this led to the predication that modulating the selective pressure of drug treatment through intermittent dosing could delay or prevent the emergence of resistant tumors. Most importantly, the preclinical data are supported by observations in vemurafenib-treated patients with melanoma providing a strong rationale for clinical testing of alternative dosing regimens. © 2013 AACR

Linking somatic genetic alterations in cancer to therapeutics.

Somatic genetic alterations provide the foundation for the evolution of human tumors as well as significant opportunity for therapeutic intervention. This review will cover the growing list of examples where somatic genetic alterations have successfully been coupled with a targeted agent resulting in positive clinical outcome. For example, recent data from randomized clinical trials support the earlier observations that EGFR mutant lung tumors are most likely to respond to EGFR kinase inhibitors, while wild-type tumors rarely respond. Emerging data indicate that this principle may also apply to such intractable diseases such as melanoma which has long been refractory to conventional chemotherapeutics

Design and synthesis of orally bioavailable benzimidazole reverse amides as pan RAF kinase inhibitors

Benzimidazole reverse amides were designed and synthesized as Pan RAF kinase inhibitors. Investigation of the structure-activity relationship of the compounds revealed that they were potent in vitro and exhibited desirable in vivo properties

Alarm tones, music and their elements: Analysis of reported waking sounds to counteract sleep inertia.

Sleep inertia is a potentially dangerous reduction in human alertness and occurs 0-4 hours after waking. The type of sound people set as their alarm for waking has been shown to reduce the effects of sleep inertia, however, the elemental musical factors that underpin these waking sounds and their relationships remain unclear. The goal of this research is to understand how a particular sound or music chosen to assist waking may counteract sleep inertia, and more specifically, what elements of these sounds may contribute to its reduction. Through an anonymous, self-report online questionnaire, fifty participants (N = 50) reported attributes of their preferred waking sound, their feeling towards the waking sound, and perceived sleep inertia after waking. This data enabled the analysis and comparison between these responses to identify statistically significant relationships. Our results did not return any significant association between sleep inertia and the reported waking sound type, nor the subject's feeling towards their sound. However, the analysis did reveal that a sound which is ranked as melodic by participants shows a significant relationship to reports of reductions in perceived sleep inertia, and in contrast, sound rated as neutral (neither unmelodic nor melodic) returns a significant relationship to the reports of increases in perceived sleep inertia. Additionally, our secondary analysis revealed that a sound rated as melodic is considered to be more rhythmic than a melodically neutral interpretation. Together these findings raise questions regarding the impact melody and rhythm may hold with respect to sleep inertia intensity. Considering that the implementation of auditory assisted awakening is a common occurrence, the musical elements of a chosen waking sound may be an area to further interrogate with respect to counteracting sleep inertia

Alarm Tones, Voice Warnings, and Musical Treatments: A Systematic Review of Auditory Countermeasures for Sleep Inertia in Abrupt and Casual Awakenings

Sleep inertia is a measurable decline in cognition some people experience upon and following awakening. However, a systematic review of the current up to date evidence of audio as a countermeasure has yet to be reported. Thus, to amend this gap in knowledge, the authors conducted this systematic review beginning with searches in three primary databases for studies published between the inception date of each journal and the year 2020. Search terms contained &ldquo;Sleep Inertia&rdquo; paired with: &ldquo;Sound&rdquo;; &ldquo;Noise&rdquo;; &ldquo;Music&rdquo;; &ldquo;Alarm&rdquo;; &ldquo;Alarm Tone&rdquo;; &ldquo;Alarm Sound&rdquo;; &ldquo;Alarm Noise&rdquo;; &ldquo;Alarm Music&rdquo;; &ldquo;Alarm Clock&rdquo;; &ldquo;Fire Alarm&rdquo;, and &ldquo;Smoke Alarm&rdquo;. From 341 study results, twelve were identified for inclusion against a priori conditions. A structured narrative synthesis approach generated three key auditory stimulus themes-(i) Noise, (ii) Emergency tone sequences; Voice Alarms and Hybrids, and (iii) Music. Across themes, participants have been assessed in two situational categories: emergency, and non-emergency awakenings. The results indicate that for children awakening in emergency conditions, a low pitch alarm or voice warnings appear to be more effective in counteracting the effects of sleep inertia than alarms with higher frequencies. For adults abruptly awakened, there is insufficient evidence to support firm conclusions regarding alarm types and voice signals. Positive results have been found in non-emergency awakenings for musical treatments in adults who preferred popular music, and alarms with melodic qualities. The results observed reflect the potential for sound, voice, and musical treatments to counteract sleep inertia post-awakening, and emphasize the requirements for further research in this domain

Auditory Countermeasures for Sleep Inertia: Exploring the Effect of Melody and Rhythm in an Ecological Context

Sleep inertia is a decline in cognition one may experience upon and following awakening. A recent study revealed that an alarm sound perceived as melodic by participants displayed a significant relationship to reports of reductions in perceived sleep inertia. This current research builds on these findings by testing the effect melody and rhythm exhibit on sleep inertia for subjects awakening in their habitual environments. Two test Groups (A and B; N = 10 each) completed an online psychomotor experiment and questionnaire in two separate test sessions immediately following awakening from nocturnal sleep. Both groups responded to a control stimulus in the first session, while in the second session, Group A experienced a melodic treatment, and Group B a rhythmic treatment. The results show that the melodic treatment significantly decreased attentional lapses, false starts, and had a significantly improved psychomotor vigilance test (PVT) performance score than the control. There was no significant result for reaction time or response speed. Additionally, no significant difference was observed for all PVT metrics between the control&ndash;rhythmic conditions. The results from this analysis support melodies&rsquo; potential to counteract symptoms of sleep inertia by the observed increase in participant vigilance following waking from nocturnal sleep

Escape from Working Poverty: Steps toward Sustainable Livelihood

Working poverty affects over half the world&#8217;s working population, yet we know remarkably little about the role of wages in transitioning toward sustainable livelihood. We develop and test a model whereby as pay approaches a living wage range, pay fairness becomes clearly associated with work&#8315;life balance; this in turn links to job satisfaction, which is a four-step process at the psychological level. We further extend this by testing a moderated mediated model, whereby income level is tested as a boundary condition. Using data from N = 873 New Zealand employees, we focus on relatively low-waged employees across three levels of income: up to $20,000,$20&#8315;40,000, and $40&#8315;60,000, with the last band straddling the New Zealand Living Wage. We find strong support for pay fairness predicting work&#8315;life balance and job satisfaction, with work&#8315;life balance mediating the relationship toward job satisfaction. In addition, we find direct effects from income to work&#8315;life balance, although not job satisfaction. Furthermore, two-way moderation is supported toward work&#8315;life balance and job satisfaction, with higher income employees reporting higher outcomes when fairness is high. The index of moderated mediation is also significantly supporting, indicating that work&#8315;life balance has a stronger mediation effect as income rises. Thus, as workers emerged from working poverty, pay fairness, and in turn work&#8315;life balance, became psychologically more salient for happiness at work, implying that a pathway to Sustainable Development Goal 8 includes at least three psychological steps, in addition to the pecuniary issue of pay: fairness, work&#8315;life balance, and job satisfaction