Fludarabine Modulates Immune Response and Extends In Vivo Survival of Adoptively Transferred CD8 T Cells in Patients with Metastatic Melanoma

Adoptive T cell therapy involving the use of ex vivo generated antigen-specific cytotoxic T lymphocytes provides a promising approach to immunotherapy. It has become increasingly apparent that anti-tumor efficacy using adoptively transferred T cells is linked to their duration of in vivo persistence and can only be achieved when combined with some form of pre-infusion patient conditioning regimen. An optimal conditioning regimen that provides a positive benefit without serious toxicities has yet to be defined. We have established a unique clinical model that allows for evaluation of a given conditioning regimen on adoptively transferred T cells in humans. In this first-in-human study (FHCRC #1796), we evaluate the use of fludarabine, an FDA-approved reagent with predictable lymphodepleting kinetics and duration of action, as a conditioning regimen that promotes homeostatic upregulation of cytokines and growth signals contributing to in vivo T cell persistence.We conducted a phase I study in patients with refractory metastatic melanoma. Patients received two infusions of a single tumor-reactive antigen-specific CTL clone expanded to 10(10)/m(2); the first infusion was given without fludarabine conditioning, and the second CTL infusion was given after a course of fludarabine (25 mg/m(2)/dayx5 days). This design permits intra-patient comparison of in vivo T cell persistence pre- and post-fludarabine. Nineteen CTL infusions were administered to ten patients. No serious toxicities were observed. Three of nine evaluable patients experienced minor response or stable disease for periods of 5.8-11.0 months with two additional patients demonstrating delayed disease stabilization. The median overall survival in this heavily pre-treated population was 9.7 months. Fludarabine led to a 2.9 fold improvement in the in vivo persistence of transferred CTL clones from a median of 4.5 days (range 0-38+) to 13.0 days (range 2-63+) (p<0.05). Fludarabine lymphodepletion increased plasma levels of the homeostatic cytokines IL-7 and IL-15. Surprisingly, fludarabine also increased the relative percentage of CD4+ T cells expressing the regulatory protein Foxp3.Lymphodepletion with fludarabine enhances transferred T cell persistence but suggest that additional improvements to optimize T cell survival and address regulatory T cells are critical in providing anti-tumor efficacy.ClinicalTrials.gov NCT00317759

Comprehensive and Integrated Genomic Characterization of Adult Soft Tissue Sarcomas

Sarcomas are a broad family of mesenchymal malignancies exhibiting remarkable histologic diversity. We describe the multi-platform molecular landscape of 206 adult soft tissue sarcomas representing 6 major types. Along with novel insights into the biology of individual sarcoma types, we report three overarching findings: (1) unlike most epithelial malignancies, these sarcomas (excepting synovial sarcoma) are characterized predominantly by copy-number changes, with low mutational loads and only a few genes (, , ) highly recurrently mutated across sarcoma types; (2) within sarcoma types, genomic and regulomic diversity of driver pathways defines molecular subtypes associated with patient outcome; and (3) the immune microenvironment, inferred from DNA methylation and mRNA profiles, associates with outcome and may inform clinical trials of immune checkpoint inhibitors. Overall, this large-scale analysis reveals previously unappreciated sarcoma-type-specific changes in copy number, methylation, RNA, and protein, providing insights into refining sarcoma therapy and relationships to other cancer types

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

Frequency and perceived effectiveness of mental health providers\u27 coping strategies during COVID-19.

There has been an emphasis on understanding the detrimental effects of coronavirus disease (COVID-19) on individuals\u27 wellbeing. Healthcare workers, including mental health providers, may experience increased emotional and behavioral health concerns to a greater degree than the general public. The objective of the present study was to examine the frequency and the perceived effectiveness of various coping strategies implemented by mental health practitioners during the COVID-19 pandemic, as well as differences across career stages (i.e., trainees versus licensed practitioners [LPs]). Survey data were collected from mental health practitioners

A broader application of the detection response task to cognitive tasks and online environments

The accurate and objective measurement of cognitive workload is important in many aspects of psychological research. The Detection Response Task (DRT) is a well-validated method for measuring cognitive workload that has been used extensively in applied tasks, for example to investigate the effects of fatigue and phone usage on driving. Given its success in applied tasks, we investigated whether the DRT could be used to measure cognitive workload in cognitive tasks more commonly used in experimental cognitive psychology, and whether this application could be extended to online environments. We had participants perform a multiple object tracking task while simultaneously performing a DRT. We manipulated the cognitive load of the multiple object tracking task by changing the number of dots to be tracked. Measurements from the DRT were sensitive to changes in the cognitive load, establishing the efficacy of the DRT for experimental cognitive tasks in lab-based situations. This sensitivity continued when applied to an online environment (with our code for the online DRT implementation being freely available at \url{https://osf.io/dc39s/}), though to a reduced extent compared to the in-lab situation, opening up the potential use of the DRT to a much greater range of tasks and situations, but suggesting that in-lab applications are best when possible

Is Multitasking Just a Form of Difficulty?

With the advancement of technologies like in-car navigation and smartphones, concerns around how cognitive functioning is influenced by ``workload'' are increasingly prevalent. Research shows that spreading effort across multiple tasks can impair cognitive abilities through an overuse of resources, and that similar overload effects arise in difficult single-task paradigms. We developed a novel lab-based extension of the Detection Response Task, which measures workload, and paired it with a Multiple Object Tracking Task to manipulate cognitive load. Load was manipulated either by changing within-task difficulty or by the addition of an extra task. Using quantitative cognitive modelling we showed that these manipulations cause similar cognitive impairments through diminished processing rates, but that the introduction of a second task tends to invoke more cautious response strategies that do not occur when only difficulty changes. We conclude that more prudence should be exercised when directly comparing multitasking and difficulty-based workload impairments, particularly when relying on measures of central tendency

A Broader Application of the Detection Response Task to Cognitive Tasks and Online Environments

Objective: The present research applied a well-established measure of cognitive workload in driving literature to an in-lab paradigm. We then extended this by comparing the in-lab version of the task to an online version. Background: The accurate and objective measurement of cognitive workload is important in many aspects of psychological research. The detection response task (DRT) is a well-validated method for measuring cognitive workload that has been used extensively in applied tasks, for example, to investigate the effects of phone usage or passenger conversation on driving, but has been used sparingly outside of this field. Method: The study investigated whether the DRT could be used to measure cognitive workload in tasks more commonly used in experimental cognitive psychology and whether this application could be extended to online environments. We had participants perform a multiple object tracking (MOT) task while simultaneously performing a DRT. We manipulated the cognitive load of the MOT task by changing the number of dots to be tracked. Results: Measurements from the DRT were sensitive to changes in the cognitive load, establishing the efficacy of the DRT for experimental cognitive tasks in lab-based situations. This sensitivity continued when applied to an online environment (our code for the online DRT implementation is freely available at https://osf.io/dc39s/), though to a reduced extent compared to the in-lab situation. Conclusion: The MOT task provides an effective manipulation of cognitive workload. The DRT is sensitive to changes in workload across a range of settings and is suitable to use outside of driving scenarios, as well as via online delivery. Application: Methodology shows how the DRT could be used to measure sources of cognitive workload in a range of human factors contexts

How is multi-tasking different from increased difficulty?

With the advancement of technologies like in-car navigation and smartphones, concerns around how cognitive functioning is influenced by "workload" are increasingly prevalent. Research shows that spreading effort across multiple tasks can impair cognitive abilities through an overuse of resources, and that similar overload effects arise in difficult single-task paradigms. We developed a novel lab-based extension of the Detection Response Task, which measures workload, and paired it with a Multiple Object Tracking Task to manipulate cognitive load. Load was manipulated either by changing within-task difficulty or by the addition of an extra task. Using quantitative cognitive modelling we showed that these manipulations cause similar cognitive impairments through diminished processing rates, but that the introduction of a second task tends to invoke more cautious response strategies that do not occur when only difficulty changes. We conclude that more prudence should be exercised when directly comparing multi-tasking and difficulty-based workload impairments, particularly when relying on measures of central tendency

Mental Health Practitioners\u27 Immediate Practical Response During the COVID-19 Pandemic: Observational Questionnaire Study.

BACKGROUND: The COVID-19 pandemic has been associated with increased psychological distress, signaling the need for increased mental health services in the context of stay-at-home policies. OBJECTIVE: This study aims to characterize how mental health practitioners have changed their practices during the pandemic. The authors hypothesize that mental health practitioners would increase tele-mental health services and that certain provider types would be better able to adapt to tele-mental health than others. METHODS: The study surveyed 903 practitioners, primarily psychologists/doctoral-level (Psych/DL) providers, social workers/master\u27s-level (SW/ML) providers, and neuropsychologists employed in academic medical centers or private practices. Differences among providers were examined using Bonferroni-adjusted chi-square tests and one-way Bonferroni-adjusted analyses of covariance. RESULTS: The majority of the 903 mental health practitioners surveyed rapidly adjusted their practices, predominantly by shifting to tele-mental health appointments (n=729, 80.82%). Whereas 80.44% (n=625) were not using tele-mental health in December 2019, only 22.07% (n=188) were not by late March or early April 2020. Only 2.11% (n=19) reported no COVID-19-related practice adjustments. Two-thirds (596/888, 67.10%) reported providing additional therapeutic services specifically to treat COVID-19-related concerns. Neuropsychologists were less likely and Psych/DL providers and SW/ML providers were more likely than expected to transition to tele-mental health (P CONCLUSIONS: The vast majority of mental health providers in this study made practice adjustments in response to COVID-19, predominantly by rapidly transitioning to tele-mental health services. Although the majority reported providing additional therapeutic services specifically to treat COVID-19-related concerns, only a small subset endorsed offering such services to medical providers. This has implications for future practical directions, as frontline workers may begin to seek mental health treatment related to the pandemic. Despite differences in tele-mental health uptake based on provider characteristics, the majority were interested in continuing to provide such services in the future. This may help to expand clinical services to those in need via tele-mental health beyond the COVID-19 pandemic

Effects of Topography-Related Puff Parameters on Carbonyl Delivery in Mainstream Cigarette Smoke

Smoking topography parameters differ substantially between individual smokers and may lead to significant variation in tobacco smoke exposure and risk for tobacco-caused diseases. However, to date, little is known regarding the impact of individual puff parameters on the delivery of many harmful smoke constituents including carbonyls. To examine this, we determined the effect of altering individual puff parameters on mainstream smoke carbonyl levels in machine-smoked reference cigarettes. Carbonyls including formaldehyde, acetaldehyde, crotonaldehyde, propionaldehyde, methyl ethyl ketone (MEK), acrolein, and acetone were determined in cigarette smoke by HPLC after derivatization with 2,4-dinitrophenylhydrazine (DNPH). Deliveries of all carbonyls were nearly two-fold greater when cigarettes were smoked according to the more intense Health Canada Intense (HCI) protocol compared to the International Organization of Standardization (ISO) method, consistent with the two-fold difference in total puff volume between methods (ISO: 280–315 mL; CI: 495–605 mL). When individual topography parameters were assessed, changes in puff volume alone had the greatest effect on carbonyl delivery as predicted with total carbonyls being strongly correlated with overall puff volume (<i>r</i>²: 0.52–0.99) regardless of how the differences in volume were achieved. All seven of the carbonyls examined showed a similar relationship with puff volume. Minor effects on carbonyl levels were observed from vent blocking and changing the interpuff interval, while effects of changing puff duration and peak flow rate were minimal. Overall, these results highlight the importance of considering topography, especially puff volume, when the toxicant delivery and potential exposure smokers receive are assessed. The lack of an impact of other behaviors, including puff intensity and duration independent of volume, indicate that factors such as temperature and peak flow rate may have minimal overall effects on carbonyl production and delivery