Establishing Meaningful Change Thresholds in Patient-Reported Outcomes Among Patients With Anaplastic Lymphoma Kinase-Positive Non-Small Cell Lung Cancer in ALTA-1L Trial.

OBJECTIVES: An earlier study from the ALTA-1L trial of patients with anaplastic lymphoma kinase-positive non-small cell lung cancer demonstrated that brigatinib produces superior health-related quality of life (QoL) outcomes over crizotinib. This study aimed to derive meaningful change thresholds (MCTs) for European Organisation for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire (QLQ)-C30 and EORTC QLQ-LC13 to refine the earlier results. METHODS: Patients from the ALTA-1L trial were administered the EORTC QLQ-C30 and EORTC QLQ-LC13 questionnaires. Responses were analyzed using anchor-based analysis, graphical analysis, distribution-based analysis, longitudinal responder analysis, and time to deterioration. RESULTS: The patient-reported outcome population comprised 262 patients who completed the EORTC QLQ-C30 at baseline and at least 1 follow-up timepoint. Both anchors (QLQ-C30 items for overall health and QoL) had correlations >0.40 or < -0.40 with all functioning domains, fatigue, pain, appetite loss, and all dyspnea scores. Within-group analysis for most scales found the derived MCT was consistent with a cutoff of 10 points for classifying individual-patient change, except for 3-item dyspnea. The probability of improvement/remaining stable was significantly greater in the brigatinib group over crizotinib for the EORTC QLQ-C30 emotional functioning, appetite loss, and constipation domains. CONCLUSIONS: This study derived MCTs for EORTC QLQ-C30 and QLQ-LC13 domains that may be applied in future studies and again demonstrated the superiority of brigatinib over crizotinib in health-related QoL outcomes in patients with non-small cell lung cancer.

Qualitative Patient Interviews to Characterize the Human Burden of Advanced Basal Cell Carcinoma Following Hedgehog Pathway Inhibitor Treatment.

INTRODUCTION: Evidence of patients' experiences of living with advanced basal cell carcinoma (aBCC) are limited, particularly after hedgehog pathway inhibitor (HHI) treatment. We explored the burden of aBCC on symptoms and patients' everyday lives post HHI treatment. METHODS: In-depth, semi-structured, approximately 1-h qualitative interviews of US patients with aBCC and prior HHI treatment were conducted. Data were assessed using thematic analysis with NVivo 1.0 software. Saturation analysis was performed to ensure all concepts were captured. RESULTS: Fifteen patients (median age, 63 years; locally advanced BCC, n = 9; metastatic BCC, n = 6) were interviewed. A patient-led conceptual model was developed from the responses using 10 symptoms and 15 impact categories (comprising emotional/psychological, physical, and social domains) identified as most commonly discussed and important to patients. Overall, reported impacts were discussed more commonly than reported symptoms. Impacts most commonly discussed were related to emotions (e.g., anxiety, worry, fear [n = 14; 93%]; low mood, depression [n = 12; 80%]) and physical function (e.g., hobbies or leisure activities [n = 13; 87%]). Symptoms most commonly discussed were fatigue and tiredness (n = 14; 93%) and itch (n = 13; 87%). Out of all reported impacts and symptoms, fatigue and tiredness (n = 7, 47%) and anxiety, worry, and fear (n = 6; 40%) were most bothersome to patients. As a descriptive exercise, participant responses were mapped to commonly used patient-reported outcome scales in aBCC clinical trials. Most expressed concepts were captured across two common measures in oncology/skin conditions (European Organization for Research and Treatment of Cancer Quality of Life-Core 30 [EORTC QLQ-C30] and Skindex-16 questionnaires), but sun avoidance and others' perception of skin cancer were not explicitly mentioned by these instruments. CONCLUSION: Patients with aBCC experienced a significant disease burden post first-line HHI therapy, including major emotional and lifestyle impacts. Accordingly, through this study, patients with aBCC highlighted a significant unmet need for second-line treatment options post HHI therapy.

Co-transducing B7H3 CAR-NK cells with the DNR preserves their cytolytic function against GBM in the presence of exogenous TGF-ß.

Cord blood (CB)-derived natural killer (NK) cells that are genetically engineered to express a chimeric antigen receptor (CAR) are an attractive off-the-shelf therapy for the treatment of cancer, demonstrating a robust safety profile in vivo. For poor prognosis brain tumors such as glioblastoma multiforme (GBM), novel therapies are urgently needed. Although CAR-T cells demonstrate efficacy in preclinical GBM models, an off-the-shelf product may exhibit unwanted side effects like graft-versus-host disease. Hence, we developed an off-the-shelf CAR-NK cell approach using a B7H3 CAR and showed that CAR-transduced NK cells have robust cytolytic activity against GBM cells in vitro. However, transforming growth factor (TGF)-ß within the tumor microenvironment has devastating effects on the cytolytic activity of both unmodified and CAR-transduced NK cells. To overcome this potent immune suppression, we demonstrated that co-transducing NK cells with a B7H3 CAR and a TGF-ß dominant negative receptor (DNR) preserves cytolytic function in the presence of exogenous TGF-ß. This study demonstrates that a novel DNR and CAR co-expression strategy may be a promising therapeutic for recalcitrant CNS tumors like GBM.

Comparable transforming growth factor beta-mediated immune suppression in ex vivo-expanded natural killer cells from cord blood and peripheral blood: implications for adoptive immunotherapy.

T cell-based therapies like genetically modified immune cells expressing chimeric antigen receptors have shown robust anti-cancer activity in vivo, especially in patients with blood cancers. However, extending this approach to an "off-the-shelf" setting can be challenging, as allogeneic T cells carry a significant risk of graft-versus-host disease (GVHD). By contrast, allogeneic natural killer (NK) cells recognize malignant cells without the need for prior antigen exposure and have been used safely in multiple cancer settings without the risk of GVHD. However, similar to T cells, NK cell function is negatively impacted by tumor-induced transforming growth factor beta (TGF-ß) secretion, which is a ubiquitous and potent immunosuppressive mechanism employed by most malignancies. Allogeneic NK cells for adoptive immunotherapy can be sourced from peripheral blood (PB) or cord blood (CB), and the authors' group and others have previously shown that ex vivo expansion and gene engineering can overcome CB-derived NK cells' functional immaturity and poor cytolytic activity, including in the presence of exogenous TGF-ß.  However, a direct comparison of the effects of TGF-ß-mediated immune suppression on ex vivo-expanded CB- versus PB-derived NK cell therapy products has not previously been performed. Here the authors show that PB- and CB-derived NK cells have distinctive gene signatures that can be overcome by ex vivo expansion. Additionally, exposure to exogenous TGF-ß results in an upregulation of inhibitory receptors on NK cells, a novel immunosuppressive mechanism not previously described. Finally, the authors provide functional and genetic evidence that both PB- and CB-derived NK cells are equivalently susceptible to TGF-ß-mediated immune suppression. The authors believe these results provide important mechanistic insights to consider when using ex vivo-expanded, TGF-ß-resistant PB- or CB-derived NK cells as novel immunotherapy agents for cancer.

The generation and application of antigen-specific T cell therapies for cancer and viral-associated disease.

Immunotherapy with antigen-specific T cells is a promising, targeted therapeutic option for patients with cancer as well as for immunocompromised patients with virus infections. In this review, we characterize and compare current manufacturing protocols for the generation of T cells specific to viral and non-viral tumor-associated antigens. Specifically, we discuss: (1) the different methodologies to expand virus-specific T cell and non-viral tumor-associated antigen-specific T cell products, (2) an overview of the immunological principles involved when developing such manufacturing protocols, and (3) proposed standardized methodologies for the generation of polyclonal, polyfunctional antigen-specific T cells irrespective of donor source. Ex vivo expanded cells have been safely administered to treat numerous patients with virus-associated malignancies, hematologic malignancies, and solid tumors. Hence, we have performed a comprehensive review of the clinical trial results evaluating the safety, feasibility, and efficacy of these products in the clinic. In summary, this review seeks to provide new insights regarding antigen-specific T cell technology to benefit a rapidly expanding T cell therapy field.

Spike-directed vaccination elicits robust spike-specific T-cell response, including to mutant strains.

Although most studies describing coronavirus disease 2019 vaccine responses have focused on antibodies, there is increasing evidence that T cells play a critical role. Here the authors evaluated T-cell responses in seronegative donors before and after vaccination to define responses to the severe acute respiratory syndrome coronavirus 2 reference strain as well as to mutations in the variant strains Alpha/B.1.1.7 and Beta/B.1.351. The authors observed enhanced T-cell responses to reference and variant spike strains post-vaccination.

Identification of novel HLA-restricted preferentially expressed antigen in melanoma peptides to facilitate off-the-shelf tumor-associated antigen-specific T-cell therapies.

BACKGROUND AIMS: Preferentially expressed antigen in melanoma (PRAME) is a cancer/testis antigen that is overexpressed in many human malignancies and poorly expressed or absent in healthy tissues, making it a good target for anti-cancer immunotherapy. Development of an effective off-the-shelf adoptive T-cell therapy for patients with relapsed or refractory solid tumors and hematological malignancies expressing PRAME antigen requires the identification of major histocompatibility complex (MHC) class I and II PRAME antigens recognized by the tumor-associated antigen (TAA) T-cell product. The authors therefore set out to extend the repertoire of HLA-restricted PRAME peptide epitopes beyond the few already characterized. METHODS: Peptide libraries of 125 overlapping 15-mer peptides spanning the entire PRAME protein sequence were used to identify HLA class I- and II-restricted epitopes. The authors also determined the HLA restriction of the identified epitopes. RESULTS: PRAME-specific T-cell products were successfully generated from peripheral blood mononuclear cells of 12 healthy donors. Ex vivo-expanded T cells were polyclonal, consisting of both CD4+ and CD8+ T cells, which elicited anti-tumor activity in vitro. Nine MHC class I-restricted PRAME epitopes were identified (seven novel and two previously described). The authors also characterized 16 individual 15-mer peptide sequences confirmed as CD4-restricted epitopes. CONCLUSIONS: TAA T cells derived from healthy donors recognize a broad range of CD4+ and CD8+ HLA-restricted PRAME epitopes, which could be used to select suitable donors for generating off-the-shelf TAA-specific T cells.

O-GlcNAc transferase is required to maintain satellite cell function.

O-GlcNAcylation is a posttranslational modification considered to be a nutrient sensor that reports nutrient scarcity or surplus. Although O-GlcNAcylation exists in a wide range of cells and/or tissues, its functional role in muscle satellite cells (SCs) remains largely unknown. Using a genetic approach, we ablated O-GlcNAc transferase (OGT), and thus O-GlcNAcylation, in SCs. We first evaluated SC function in vivo using a muscle injury model and found that OGT deficient SCs had compromised capacity to repair muscle after an acute injury compared with the wild-type SCs. By tracing SC cycling rates in vivo using the doxycycline-inducible H2B-GFP mouse model, we found that SCs lacking OGT cycled at lower rates and reduced in abundance with time. Additionally, the self-renewal ability of OGT-deficient SCs after injury was decreased compared to that of the wild-type SCs. Moreover, in vivo, in vitro, and ex vivo proliferation assays revealed that SCs lacking OGT were incapable of expanding compared with their wild-type counterparts, a phenotype that may be explained, at least in part, by an HCF1-mediated arrest in the cell cycle. Taken together, our findings suggest that O-GlcNAcylation plays a critical role in the maintenance of SC health and function in normal and injured skeletal muscle.

NK Cell Adoptive Immunotherapy of Cancer: Evaluating Recognition Strategies and Overcoming Limitations.

Natural killer (NK) cells, the primary effector cells of the innate immune system, utilize multiple strategies to recognize tumor cells by (1) detecting the presence of activating receptor ligands, which are often upregulated in cancer; (2) targeting cells that have a loss of major histocompatibility complex (MHC); and (3) binding to antibodies that bind to tumor-specific antigens on the tumor cell surface. All these strategies have been successfully harnessed in adoptive NK cell immunotherapies targeting cancer. In this review, we review the applications of NK cell therapies across different tumor types. Similar to other forms of immunotherapy, tumor-induced immune escape and immune suppression can limit NK cell therapies' efficacy. Therefore, we also discuss how these limitations can be overcome by conferring NK cells with the ability to redirect their tumor-targeting capabilities and survive the immune-suppressive tumor microenvironment. Finally, we also discuss how future iterations can benefit from combination therapies with other immunotherapeutic agents.

SARS-CoV-2-specific T cells are rapidly expanded for therapeutic use and target conserved regions of the membrane protein.

T-cell responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been described in recovered patients, and may be important for immunity following infection and vaccination as well as for the development of an adoptive immunotherapy for the treatment of immunocompromised individuals. In this report, we demonstrate that SARS-CoV-2-specific T cells can be expanded from convalescent donors and recognize immunodominant viral epitopes in conserved regions of membrane, spike, and nucleocapsid. Following in vitro expansion using a good manufacturing practice-compliant methodology (designed to allow the rapid translation of this novel SARS-CoV-2 T-cell therapy to the clinic), membrane, spike, and nucleocapsid peptides elicited interferon-γ production, in 27 (59%), 12 (26%), and 10 (22%) convalescent donors (respectively), as well as in 2 of 15 unexposed controls. We identified multiple polyfunctional CD4-restricted T-cell epitopes within a highly conserved region of membrane protein, which induced polyfunctional T-cell responses, which may be critical for the development of effective vaccine and T-cell therapies. Hence, our study shows that SARS-CoV-2 directed T-cell immunotherapy targeting structural proteins, most importantly membrane protein, should be feasible for the prevention or early treatment of SARS-CoV-2 infection in immunocompromised patients with blood disorders or after bone marrow transplantation to achieve antiviral control while mitigating uncontrolled inflammation.

Dual effects of obesity on satellite cells and muscle regeneration.

Obesity is a complex metabolic disorder that often leads to a decrease in insulin sensitivity, chronic inflammation, and overall decline in human health and well-being. In mouse skeletal muscle, obesity has been shown to impair muscle regeneration after injury; however, the mechanism underlying these changes has yet to be determined. To test whether there is a negative impact of obesity on satellite cell (SC) decisions and behaviors, we fed C57BL/6 mice normal chow (NC, control) or a high-fat diet (HFD) for 10 weeks and performed SC proliferation and differentiation assays in vitro. SCs from HFD mice formed colonies with smaller size (p < .001) compared to those from NC mice, and this decreased proliferation was confirmed (p < .05) by BrdU incorporation. Moreover, in vitro assays showed that HFD SCs exhibited diminished (p < .001) fusion capacity compared to NC SCs. In single fiber explants, a higher ratio of SCs experienced apoptotic events (p < .001) in HFD mice compared to that of NC-fed mice. In vivo lineage tracing using H2B-GFP mice showed that SCs from HFD treatment also cycled faster (p < .001) than their NC counterparts. In spite of all these autonomous cellular effects, obesity as triggered by high-fat feeding did not significantly impair muscle regeneration in vivo, as reflected by the comparable cross-sectional area (p > .05) of the regenerating fibers in HFD and NC muscles, suggesting that other factors may mitigate the negative impact of obesity on SCs properties.

The effect of perceived appearance judgements on psychological and biological stress processes across adulthood.

Social self-preservation theory posits that stress is experienced when an aspect of an individual's identity has the potential to be negatively evaluated. Appearance is a central part of identity; however, little research has examined whether perceived appearance judgements are a source of social-evaluative stress. In addition, stress may be an explanatory link in the association between appearance perceptions and depressive symptoms. This study examined whether perceived appearance judgements were associated with increased stress and greater depressive symptoms among adults. Study 1 examined the associations between self-reported appearance judgements and cortisol stress responses in response to a laboratory stressor (Trier Social Stress Test) among 71 individuals aged 18-65. Study 2 assessed self-reported appearance judgements and depressive symptoms among 498 adults ages 18-65 via an online survey data collection. Appearance judgement was associated with a stronger cortisol response, higher self-reported stress, and greater depressive symptoms. Stress mediated all associations between appearance judgements and depressive symptoms and neither age nor gender moderated these associations. The findings suggest that appearance judgements contribute to psychological and biological stress processes and demonstrated that stress mediated the association between appearance judgements and depressive symptoms.

Comparison group matters for chronic stress effects of subjective social status.

Inconsistent associations between health and measures of subjective social status compared to one's community suggest that how people define community may matter. This study broke down community into status among neighborhood and friends/family to assess the impact of each domain on chronic stress in individuals differing in socioeconomic status (18 employed and 18 unemployed individuals). The findings suggest that for ratings of subjective social status, the social and physical proximity of the reference group matters. Specifically, neighborhood status was affected by unemployment, while friends/family status was associated with perceived stress, emphasizing the importance of the comparison group in assessing subjective status.

Low subjective social status in the police is linked to health-relevant changes in diurnal salivary alpha-amylase activity in Swiss police officers.

The objective of this study was to assess basal autonomic nervous system (ANS) activity as a pathway linking subjective social status to health in a high-demand work environment. It was hypothesized that officers with a lower status experienced more chronic stress (higher basal ANS activity) and that chronic stress was related to more health problems. Fifty-six male and female Swiss police officers self-reported on subjective social status (country, community, friends, police) and their health (depression, post-traumatic stress, physical symptoms) and collected 12 saliva samples over two days for basal α-amylase activation (sAA) assessment. Multilevel regression analyses revealed that subjective social status in the police and physical symptoms explained a significant part of the variance in diurnal sAA activity patterns. The current findings support the idea that more narrowly defined subjective social status may be more closely linked to biological stress mechanisms. Additionally, sAA activity was specifically related to physical, but not mental health problems. These results suggest that subjective social status referencing one's work environment may be a promising early indicator of health-relevant changes in stress-related physiological systems.

Leisure activities are linked to mental health benefits by providing time structure: comparing employed, unemployed and homemakers.

BACKGROUND: Unemployment has consistently been linked to negative mental health outcomes, emphasising the need to characterise the underlying mechanisms. The current study aimed at testing whether compared with other employment groups, fewer leisure activities observed in unemployment may contribute to elevated risk for negative mental health via loss of time structure. METHODS: Depressive symptoms (Center for Epidemiologic Studies Depression), leisure activities (exercise, self-focused, social), and time structure (Time Structure Questionnaire (TSQ)) were assessed cross-sectionally in 406 participants (unemployed=155, employed=140, homemakers=111) recruited through Amazon Mechanical Turk. RESULTS: Controlling for gender and age, structural equation modelling revealed time structure partially (employed, homemakers) and fully (unemployed) mediated the relationship between leisure activities and depressive symptoms. With the exception of differential effects for structured routines, all other TSQ factors (sense of purpose, present orientation, effective organisation and persistence) contributed significantly to all models. CONCLUSIONS: These findings support the idea that especially for the unemployed, leisure activities impose their mental health benefits through increasing individuals' perception of spending their time effectively. Social leisure activities that provide a sense of daily structure may thereby be a particularly promising low-cost intervention to improve mental health in this population.

Differential Links Between Leisure Activities and Depressive Symptoms in Unemployed Individuals.

OBJECTIVE: Unemployment has consistently been linked to an elevated risk for depression. Exercise, specifically leisure-based physical activities, has received increasing attention as alternative treatment options. However, because leisure activities are pursued during discretionary time, it is unclear if the mental health benefits of physical and leisure activities apply during times of unemployment as well. METHOD: Depressive symptoms and participation in recreational activities were assessed in 142 employed and 158 unemployed participants (age = 34 ± 11 years; male = 150). RESULTS: Independent of employment status, all recreational activities were inversely associated with depressive symptoms. However, social (employed: ηp (2) = .21; unemployed: ηp (2) = .11) and self-focused (employed: ηp (2) = .19; unemployed: ηp (2) = .10) recreational activities were more strongly related to depressive symptoms than exercise (employed: ηp (2) = .12; unemployed: ηp (2) > .05). CONCLUSION: These findings highlight the strong mental health associations of recreational activities and suggest that, particularly for unemployed individuals, promoting recreational activities, rather than exercise, may leverage the stronger negative relationship with risk of depression.

Health in police officers: Role of risk factor clusters and police divisions.

OBJECTIVE: Law enforcement is a stressful occupation associated with significant health problems. To date, most studies have focused on one specific factor or one domain of risk factors (e.g., organizational, personal). However, it is more likely that specific combinations of risk factors are differentially health relevant and further, depend on the area of police work. METHODS: A self-selected group of officers from the criminal, community, and emergency division (N = 84) of a Swiss state police department answered questionnaires assessing personal and organizational risk factors as well as mental and physical health indicators. RESULTS: In general, few differences were observed across divisions in terms of risk factors or health indicators. Cluster analysis of all risk factors established a high-risk and a low-risk cluster with significant links to all mental health outcomes. Risk cluster-by-division interactions revealed that, in the high-risk cluster, Emergency officers reported fewer physical symptoms, while community officers reported more posttraumatic stress symptoms. Criminal officers in the high-risk cluster tended to perceived more stress. Finally, perceived stress did not mediate the relationship between risk clusters and posttraumatic stress symptoms. CONCLUSION: In summary, our results support the notion that police officers are a heterogeneous population in terms of processes linking risk factors and health indicators. This heterogeneity thereby appeared to be more dependent on personal factors and individuals' perception of their own work conditions than division-specific work environments. Our findings further suggest that stress-reduction interventions that do not target job-relevant sources of stress may only show limited effectiveness in reducing health risks associated with police work.

Cortisol-dependent stress effects on cell distribution in healthy individuals and individuals suffering from chronic adrenal insufficiency.

Chronic adrenal insufficiency (CAI) is characterized by a lack of glucocorticoid and mineralocorticoid production due to destroyed adrenal cortex cells. However, elevated cortisol secretion is thought to be a central part in a well-orchestrated immune response to stress. This raises the question to what extent lack of cortisol in CAI affects stress-related changes in immune processes. To address this question, 28 CAI patients (20 females) and 18 healthy individuals (11 females) (age: 44.3 ± 8.4 years) were exposed to a psychosocial stress test (Trier Social Stress Test: TSST). Half the patients received a 0.03 mg/kg body weight injection of hydrocortisone (HC) post-TSST to mimic a healthy cortisol stress response. Catecholamines and immune cell composition were assessed in peripheral blood and free cortisol measured in saliva collected before and repeatedly after TSST. CAI patients showed norepinephrine (NE) stress responses similar to healthy participants, however, epinephrine (E) as well as cortisol levels were significantly lower. HC treatment post-TSST resulted in cortisol increases comparable to those observed in healthy participants (interaction effects--NE: F=1.05, p=.41; E: F=2.56, p=.045; cortisol: F=13.28, p<.001). Healthy individuals showed the expected pattern of stress-related early lymphocyte increase with subsequent decrease below baseline. The opposite pattern was observed in granulocytes. While exhibiting a similar initial increase, lymphocytes kept increasing over the following 2h in untreated patients. HC treatment buffered this effect (interaction effects--lymphocyte%: F=7.31, p<.001; granulocyte%: F=7.71, p<.001). Using CAI in humans as a model confirms cortisol's central involvement in post-stress lymphocyte migration from blood into immune-relevant body compartments. As such, future studies should investigate whether psychosocial stress exposure may put CAI patients at an increased health risk due to attenuated immune responses to pathogens.

Perceived appearance judgments moderate the biological stress effects of social exchanges.

Social relationships are generally thought of as beneficial. However, the present study set out to test the hypothesis that for individuals who perceive others to judge their appearance negatively, daily social interactions can also be a source of stress. Indeed when assessing 38 young adults, we found that both more incidences of negative exchanges reported during the past month as well as perceived negative appearance judgments by others were associated with more self-reported stress. Interestingly, however, for individuals with low attribution body esteem, higher numbers of positive social exchanges during the past month were related to health-relevant changes in biological markers of chronic stress as well. The same was true for individuals with high attribution body esteem who reported to experience only very few positive exchanges. As such, these findings go beyond the initial focus on low body esteem and negative social exchanges and introduce high body esteem as well as daily positive exchanges as potential health risk factors.