Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells.

Checkpoint inhibitors have revolutionized cancer treatment. However, only a minority of patients respond to these immunotherapies. Here, we report that blocking the inhibitory NKG2A receptor enhances tumor immunity by promoting both natural killer (NK) and CD8+ T cell effector functions in mice and humans. Monalizumab, a humanized anti-NKG2A antibody, enhanced NK cell activity against various tumor cells and rescued CD8+ T cell function in combination with PD-x axis blockade. Monalizumab also stimulated NK cell activity against antibody-coated target cells. Interim results of a phase II trial of monalizumab plus cetuximab in previously treated squamous cell carcinoma of the head and neck showed a 31% objective response rate. Most common adverse events were fatigue (17%), pyrexia (13%), and headache (10%). NKG2A targeting with monalizumab is thus a novel checkpoint inhibitory mechanism promoting anti-tumor immunity by enhancing the activity of both T and NK cells, which may complement first-generation immunotherapies against cancer.

GM-CSF-activated human dendritic cells promote type 1 T follicular helper cell polarization in a CD40-dependent manner.

T follicular helper (Tfh) cells regulate humoral responses and present a marked phenotypic and functional diversity. Type 1 Tfh (Tfh1) cells were recently identified and associated with disease severity in infection and autoimmune diseases. The cellular and molecular requirements to induce human Tfh1 differentiation are not known. Here, using single-cell RNA sequencing (scRNAseq) and protein validation, we report that human blood CD1c+ dendritic cells (DCs) activated by GM-CSF (also known as CSF2) drive the differentiation of naive CD4+ T cells into Tfh1 cells. These Tfh1 cells displayed typical Tfh molecular features, including high levels of PD-1 (encoded by PDCD1), CXCR5 and ICOS. They co-expressed BCL6 and TBET (encoded by TBX21), and secreted large amounts of IL-21 and IFN-γ (encoded by IFNG). Mechanistically, GM-CSF triggered the emergence of two DC sub-populations defined by their expression of CD40 and ICOS ligand (ICOS-L), presenting distinct phenotypes, morphologies, transcriptomic signatures and functions. CD40High ICOS-LLow DCs efficiently induced Tfh1 differentiation in a CD40-dependent manner. In patients with mild COVID-19 or latent Mycobacterium tuberculosis infection, Tfh1 cells were positively correlated with a CD40High ICOS-LLow DC signature in scRNAseq of peripheral blood mononuclear cells or blood transcriptomics, respectively. Our study uncovered a novel CD40-dependent Tfh1 axis with potential physiopathological relevance to infection. This article has an associated First Person interview with the first author of the paper.

The predictive power of insomnia symptoms on other aspects of mental health during the COVID-19 pandemic: a longitudinal study.

Symptoms of insomnia are an important risk factor for the development of mental disorders, especially during stressful life periods such as the coronavirus disease 2019 (COVID-19) pandemic. However, up to now, most studies have used cross-sectional data, and the prolonged impact of insomnia symptoms during the pandemic on later mental health remains unclear. Therefore, we investigated insomnia symptoms as a predictor of other aspects of mental health across 6 months, with altogether seven assessments (every 30 days, t0-t6), in a community sample (N = 166-267). Results showed no mean-level increase of insomnia symptoms and/or deterioration of mental health between baseline assessment (t0) and the 6- month follow-up (t6). As preregistered, higher insomnia symptoms (between persons) across all time points predicted reduced mental health at the 6-month follow-up. Interestingly, contrary to our hypothesis, higher insomnia symptoms at 1 month, within each person (i.e., compared to that person's symptoms at other time points), predicted improved rather than reduced aspects of mental health 1 month later. Hence, we replicated the predictive effect of averagely increased insomnia symptoms on impaired later mental health during the COVID-19 pandemic. However, we were surprised that increased insomnia symptoms at 1 month predicted aspects of improved mental health 1 month later. This unexpected effect might be specific for our study population and a consequence of our study design. Overall, increased insomnia symptoms may have served as a signal to engage in, and successfully implement, targeted countermeasures, which led to better short-term mental health in this healthy sample.

ß-Boronic Acid-Substituted Bodipy Dyes for Fluorescence Anisotropy Analysis of Carbohydrate Binding.

Boronic acids are widely used for labeling catechols and carbohydrates in analytical (bio)chemistry due to their high binding affinities for diols. Here, we present two asymmetrically substituted Bodipy dyes with a boronic acid at the ß-position (BBB). We present a green-emitting BBB, gBBB, and, by expanding the conjugated system of the Bodipy core at α-position, a red-emitting rBBB. Especially, gBBB shows a bathochromic shift of the electronic spectra upon binding to saccharides and polyols, whereas the fluorescence lifetime of rBBB is more sensitive to hydroxy-ligand binding. Moreover, gBBB constantly shows higher binding affinities than rBBB, reaching Kb ≈ 103 M-1 at pH 8.5 for fructose. Finally, time-resolved fluorescence anisotropy allows to distinguish the number of saccharide units of oligosaccharides as the bond along the transition dipole moment ensures that the fluorescence anisotropy only decays due to the rotational diffusion of labeled carbohydrates. ß-substituted BODIPY dyes are, thus, foreseen as fluorescence anisotropy labels for macromolecule sizing, where conventional fluorophores fail to discriminate due to the chemical similarity of recognition sites.

Nanoparticle therapy for head and neck cancers.

PURPOSE OF REVIEW: The current review focuses on the therapeutic use of nanoparticles in head and neck cancer (HNC), highlighting nanoparticles at the most advanced clinical development stages. RECENT FINDINGS: Literature review covers the three main approaches for therapeutic use of nanoparticles in HNC: first, enhancing radiotherapy effect; second, performing targeted delivery of chemotherapy, immunotherapy, or genome editing molecules; third, photothermal therapy. SUMMARY: Nanoparticles are spherical nanoscale objects that have application in cancer therapies. Nanoparticles have diverse and often composite structure composition to ensure their function, increase their bioavailability in tumor tissues, and decrease off-target effects, sometimes by means of activating internal or external stimuli. Hafnium oxide nanoparticles are being tested in phase I to III trials for radiotherapy enhancement. Nanoparticle-based delivery of paclitaxel, cisplatin, and of the immune activator CpG-A DNA is being evaluated in phase II trials. No nanoparticle is currently approved for HNC treatment.

Measuring motivational relationship processes in experience sampling: A reliability model for moments, days, and persons nested in couples.

The investigation of within-person process models, often done in experience sampling designs, requires a reliable assessment of within-person change. In this paper, we focus on dyadic intensive longitudinal designs where both partners of a couple are assessed multiple times each day across several days. We introduce a statistical model for variance decomposition based on generalizability theory (extending P. E. Shrout & S. P. Lane, 2012), which can estimate the relative proportion of variability on four hierarchical levels: moments within a day, days, persons, and couples. Based on these variance estimates, four reliability coefficients are derived: between-couples, between-persons, within-persons/between-days, and within-persons/between-moments. We apply the model to two dyadic intensive experience sampling studies (n1 = 130 persons, 5 surveys each day for 14 days, ≥ 7508 unique surveys; n2 = 508 persons, 5 surveys each day for 28 days, ≥ 47764 unique surveys). Five different scales in the domain of motivational processes and relationship quality were assessed with 2 to 5 items: State relationship satisfaction, communal motivation, and agentic motivation; the latter consists of two subscales, namely power and independence motivation. Largest variance components were on the level of persons, moments, couples, and days, where within-day variance was generally larger than between-day variance. Reliabilities ranged from .32 to .76 (couple level), .93 to .98 (person level), .61 to .88 (day level), and .28 to .72 (moment level). Scale intercorrelations reveal differential structures between and within persons, which has consequences for theory building and statistical modeling.

Plasmacytoid pre-dendritic cells (pDC): from molecular pathways to function and disease association.

Plasmacytoid pre-dendritic cells (pDC) are a specialized DC population with a great potential to produce large amounts of type I interferon (IFN). pDC are involved in the initiation of antiviral immune responses through their interaction with innate and adaptive immune cell populations. In a context-dependent manner, pDC activation can induce their differentiation into mature DC able to induce both T cell activation or tolerance. In this review, we described pDC functions during immune responses and their implication in the clearance or pathogenicity of human diseases during infection, autoimmunity, allergy and cancer. We discuss recent advances in the field of pDC biology and their implication for future studies.

Measuring Implicit Motives with the Picture Story Exercise (PSE): Databases of Expert-Coded German Stories, Pictures, and Updated Picture Norms.

We present two openly accessible databases related to the assessment of implicit motives using Picture Story Exercises (PSEs): (a) A database of 183,415 German sentences, nested in 26,389 stories provided by 4,570 participants, which have been coded by experts using Winter's coding system for the implicit affiliation/intimacy, achievement, and power motives, and (b) a database of 54 classic and new pictures which have been used as PSE stimuli. Updated picture norms are provided which can be used to select appropriate pictures for PSE applications. Based on an analysis of the relations between raw motive scores, word count, and sentence count, we give recommendations on how to control motive scores for story length, and validate the recommendation with a meta-analysis on gender differences in the implicit affiliation motive that replicates existing findings. We discuss to what extent the guiding principles of the story length correction can be generalized to other content coding systems for narrative material. Several potential applications of the databases are discussed, including (un)supervised machine learning of text content, psychometrics, and better reproducibility of PSE research.

Surface Preparation for Single-Molecule Chemistry.

Immobilization procedures, intended to enable prolonged observation of single molecules by fluorescence microscopy, may generate heterogeneous microenvironments, thus inducing heterogeneity in the molecular behavior. On that account, we propose a straightforward surface preparation procedure for studying chemical reactions on the single-molecule level. Sensor fluorophores were developed, which exhibit dual-emissive characteristics in a homogeneously catalyzed showcase reaction. These molecules undergo a shift of fluorescence wavelength of about 100 nm upon Pd(0)-induced deallylation in the Tsuji-Trost reaction, allowing for separate visualization of the starting material and product. Whereas a simultaneous immobilization of dye and inert silane leads to strongly polydisperse reaction kinetics, a consecutive immobilization routine with deposition of dye molecules as the last step provides substrates underlying the kinetics of ensemble experiments. Also, the found kinetics are unaffected by the chemical variation of inert silanes, nearly uniform, and therefore well reproducible. Additional parameters like photostability, signal-to-noise ratio, dye-molecule density, and spatial distribution of dye molecules are, as well, hardly affected by surface modification in the successive immobilization scheme.

Acellular fish skin matrix on thin-skin graft donor sites: a preliminary study.

OBJECTIVE: Surgery for head and neck cancer often requires free flap reconstructions, whose harvesting site often requires a thin-skin graft. Wounds from the thin-skin donor site are comparable to an intermediate or deep second-degree burn. This is uncomfortable and can lead to complications such as a long healing time, local infections and pain. Since they are reproducible, these wounds may serve as a model for an objective assessment of new healing medical devices. The acellular fish skin matrix is a new medical device designed to improve healing quality and time. METHODS: We compared the outcomes between standard procedure and the use of this matrix placed on the split-thickness skin graft (STSG) donor site, in patients operated on in our centre for radial forearm free flap reconstruction for head and neck wounds. RESULTS: There were 21 patients included. The healing time was halved when using the acellular fish skin matrix, from 68 to 32 days on average. Acellular fish skin matrix reduced pain levels and local infection. The visual analogue pain scale (VAS) was ≥3 at five days (p=0.0034) and infection rate reduced from 60% to 0% (p=0.0039). CONCLUSION: These results are extremely encouraging. However, it is important to take into account the relatively high cost of this matrix for its future indications. A larger study including an overall cost estimation and an assessment on different wound types would be interesting, to better target the indications of the acellular fish skin matrix.

Combating the Health Risks of Sedentary Behavior in the Contemporary Radiology Reading Room.

OBJECTIVE: We detail the association of sedentary behavior with a variety of health problems and provide the radiologist with a number of simple activities and techniques that can improve overall health while still meeting the productivity demands of a high-volume practice. Although these techniques are well known and recognized in the fitness and nutrition literature, they are not widely used in the radiology reading room. The computer- and workstation-based work routinely performed by diagnostic radiologists typically occurs in the seated position, leading to more than 8 hours per day of sitting. Studies have found that even for those who exercise regularly, spending increased time sitting can negate the healthful effects of exercise. Time spent in a seated or sedentary position leads to slowing of one's metabolism, with negative resultant effects. CONCLUSION: The concept of nonexercise activity thermogenesis (NEAT) will be described, with examples given of how to burn more calories while at work and, therefore, improve the health of the diagnostic radiologist. NEAT refers to the energy expended during activities of daily living, excluding sportlike or intentional exercise. The concept of NEAT must be understood by radiologists, because it allows the development of multiple strategies to combat the ill effects of sitting while working. Adding intermittent movement and stretching exercises throughout the day can stimulate metabolism. An understanding of the association of sedentary work behavior with a number of health risks is crucial for radiologists so that they can implement basic changes into their work routine, allowing them to increase activity to address and avoid these potential health hazards.

Safety and reliability of the internal jugular vein for venous anastomoses in head and neck oncological reconstruction: A retrospective study.

This study aimed to assess the efficacy of utilizing the internal jugular vein (IJV) as the primary recipient site for venous anastomoses in head and neck oncological reconstruction. Patients who underwent a free flap reconstruction of the head and neck were retrospectively included. Venous anastomoses were preferentially performed less than 1 cm from the IJV, either end-to-side (EtS) on the IJV, or end-to-end (EtE) on the origin of the thyrolingofacial venous (TLF) trunk. When the pedicle length was insufficient to reach the IJV, anastomoses were performed EtE to a size-matched cervical vein. Of the 246 venous anastomoses, 216 (87.8%) were performed less than 1 cm from the IJV, including 150 EtS on the IJV (61.0%), and 66 EtE on the TLF trunk (26.8%). Thirty veins (12.1%) were anastomosed EtE on other cervical veins more than 1 cm from the IJV. Two venous thromboses occurred (0.9%) and were successfully managed after revision surgery. There was no evidence of an increased thrombosis rate in high-risk or pre-irradiated patients. These findings suggest that the internal jugular vein is safe and reliable as a first-choice recipient vessel for free flap transfers in head and neck oncological reconstruction.

Tumor draining lymph nodes connected to cold triple-negative breast cancers are characterized by Th2-associated microenvironment.

Tumor draining lymph nodes (TDLN) represent a key component of the tumor-immunity cycle. There are few studies describing how TDLNs impact lymphocyte infiltration into tumors. Here we directly compare tumor-free TDLNs draining "cold" and "hot" human triple negative breast cancers (TDLNCold and TDLNHot). Using machine-learning-based self-correlation analysis of immune gene expression, we find unbalanced intranodal regulations within TDLNCold. Two gene pairs (TBX21/GATA3-CXCR1) with opposite correlations suggest preferential priming of T helper 2 (Th2) cells by mature dendritic cells (DC) within TDLNCold. This is validated by multiplex immunofluorescent staining, identifying more mature-DC-Th2 spatial clusters within TDLNCold versus TDLNHot. Associated with this Th2 priming preference, more IL4 producing mast cells (MC) are found within sinus regions of TDLNCold. Downstream, Th2-associated fibrotic TME is found in paired cold tumors with increased Th2/T-helper-1-cell (Th1) ratio, upregulated fibrosis growth factors, and stromal enrichment of cancer associated fibroblasts. These findings are further confirmed in a validation cohort and public genomic data. Our results reveal a potential role of IL4+ MCs within TDLNs, associated with Th2 polarization and reduced immune infiltration into tumors.

International guidelines for intratumoral and intranodal injection of NBTXR3 nanoparticles in head and neck cancers.

BACKGROUND: An international multidisciplinary panel of experts aimed to provide consensus guidelines describing the optimal intratumoral and intranodal injection of NBTXR3 hafnium oxide nanoparticles in head and neck squamous cell carcinoma (HNSCC) of the oral cavity, oropharynx, and cervical lymph nodes and to review data concerning safety, feasibility, and procedural aspects of administration. METHODS: The Delphi method was used to determine consensus. A 4-member steering committee and a 10-member monitoring committee wrote and revised the guidelines, divided into eight sections. An independent 3-member reading committee reviewed the recommendations. RESULTS: After two rounds of voting, strong consensus was obtained on all recommendations. Intratumoral and intranodal injection was deemed feasible. NBTXR3 volume calculation, choice of patients, preparation and injection procedure, potential side effects, post injection, and post treatment follow-up were described in detail. CONCLUSIONS: Best practices for the injection of NBTXR3 were defined, thus enabling international standardization of intratumoral nanoparticle injection.

Breastfeeding of preterm newborns in a neonate hospitalization unit.

OBJECTIVE: To analyze the factors associated with breastfeeding of preterm infants at discharge. METHOD: Cross-sectional study with newborns with gestational age <37 weeks, admitted to a university hospital. Data obtained from the medical records of 180 participants, from August/2019 to August/2020. To assess an association between categorical variables, Pearson's chi-square and Fisher's exact tests were used. The significance level adopted was 5% (p ≤ 0.05). RESULTS: Mean gestational age was 32.8 ± 2.7 weeks and mean birth weight was 1,890 grams ± 682 grams. During hospitalization, (n=166) 28.3% received predominantly breast milk. At discharge, (n=164) 84.1% received breast milk and, of these, 2.4% were exclusively breastfed. Breastfeeding at discharge was associated with gestational age ≥ 33.5 weeks, higher weight at birth, and shorter hospitalization. CONCLUSION: The study showed that during hospitalization, about a third of the participants were breastfed. However, at the time of discharge, there was a predominance of breastfeeding in most cases, and the associated factors were higher weight at birth and shorter hospital stay.

Large-scale analysis of cell-cell communication reveals angiogenin-dependent tumor progression in clear cell renal cell carcinoma.

Cellular crosstalk in the tumor microenvironment (TME) is still largely uncharacterized, while it plays an essential role in shaping immunosuppression or anti-tumor response. Large-scale analyses are needed to better decipher cell-cell communication in cancer. In this work, we used original and publicly available single-cell RNA sequencing (scRNAseq) data to characterize in-depth the communication networks in human clear cell renal cell carcinoma (ccRCC). We identified 50 putative communication channels specifically used by cancer cells to interact with other cells, including two novel angiogenin-mediated interactions. Expression of angiogenin and its receptors was validated at the protein level in primary ccRCC. Mechanistically, angiogenin enhanced ccRCC cell line proliferation and down-regulated secretion of IL-6, IL-8, and MCP-1 proinflammatory molecules. This study provides novel biological insights into molecular mechanisms of ccRCC, and suggests angiogenin and its receptors as potential therapeutic targets in clear cell renal cancer.

Meta-Analysis of Human Cancer Single-Cell RNA-Seq Datasets Using the IMMUcan Database.

The development of single-cell RNA sequencing (scRNA-seq) technologies has greatly contributed to deciphering the tumor microenvironment (TME). An enormous amount of independent scRNA-seq studies have been published representing a valuable resource that provides opportunities for meta-analysis studies. However, the massive amount of biological information, the marked heterogeneity and variability between studies, and the technical challenges in processing heterogeneous datasets create major bottlenecks for the full exploitation of scRNA-seq data. We have developed IMMUcan scDB (https://immucanscdb.vital-it.ch), a fully integrated scRNA-seq database exclusively dedicated to human cancer and accessible to nonspecialists. IMMUcan scDB encompasses 144 datasets on 56 different cancer types, annotated in 50 fields containing precise clinical, technological, and biological information. A data processing pipeline was developed and organized in four steps: (i) data collection; (ii) data processing (quality control and sample integration); (iii) supervised cell annotation with a cell ontology classifier of the TME; and (iv) interface to analyze TME in a cancer type-specific or global manner. This framework was used to explore datasets across tumor locations in a gene-centric (CXCL13) and cell-centric (B cells) manner as well as to conduct meta-analysis studies such as ranking immune cell types and genes correlated to malignant transformation. This integrated, freely accessible, and user-friendly resource represents an unprecedented level of detailed annotation, offering vast possibilities for downstream exploitation of human cancer scRNA-seq data for discovery and validation studies. SIGNIFICANCE: The IMMUcan scDB database is an accessible supportive tool to analyze and decipher tumor-associated single-cell RNA sequencing data, allowing researchers to maximally use this data to provide new insights into cancer biology.

Selective STING stimulation in dendritic cells primes antitumor T cell responses.

T cells that recognize tumor antigens are crucial for mounting antitumor immune responses. Induction of antitumor T cells in immunogenic tumors depends on STING, the intracellular innate immune receptor for cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) and related cyclic dinucleotides (CDNs). However, the optimal way to leverage STING activation in nonimmunogenic tumors is still unclear. Here, we show that cGAMP delivery by intratumoral injection of virus-like particles (cGAMP-VLP) led to differentiation of circulating tumor-specific T cells, decreased tumor regulatory T cells (Tregs), and antitumoral responses that synergized with PD1 blockade. By contrast, intratumoral injection of the synthetic CDN ADU-S100 led to tumor necrosis and systemic T cell activation but simultaneously depleted immune cells from injected tumors and induced minimal priming of circulating tumor-specific T cells. The antitumor effects of cGAMP-VLP required type 1 conventional dendritic cells (cDC1), whereas ADU-S100 eliminated cDC1 from injected tumors. cGAMP-VLP preferentially targeted STING in dendritic cells at a 1000-fold smaller dose than ADU-S100. Subcutaneous administration of cGAMP-VLP showed synergy when combined with PD1 blockade or a tumor Treg-depleting antibody to elicit systemic tumor-specific T cells and antitumor activity, leading to complete and durable tumor eradication in the case of tumor Treg depletion. These findings show that cell targeting of STING stimulation shapes the antitumor T cell response and identify a therapeutic strategy to enhance T cell-targeted immunotherapy.

Randomized phase II study of preoperative afatinib in untreated head and neck cancers: predictive and pharmacodynamic biomarkers of activity.

There is no strong and reliable predictive biomarker in head and neck squamous cell carcinoma (HNSCC) for EGFR inhibitors. We aimed to identify predictive and pharmacodynamic biomarkers of efficacy of afatinib, a pan-HER tyrosine kinase inhibitor, in a window-of-opportunity trial (NCT01415674). Multi-omics analyses were carried out on pre-treatment biopsy and surgical specimen for biological assessment of afatinib activity. Sixty-one treatment-naïve and operable HNSCC patients were randomised to afatinib 40 mg/day for 21-28 days versus no treatment. Afatinib produced a high rate of metabolic response. Responders had a higher expression of pERK1/2 (P = 0.02) and lower expressions of pHER4 (P = 0.03) and pRB1 (P = 0.002) in pre-treatment biopsy compared to non-responders. At the cellular level, responders displayed an enrichment of tumor-infiltrating B cells under afatinib (P = 0.02). At the molecular level, NF-kappa B signaling was over-represented among upregulated genes in non-responders (P < 0.001; FDR = 0.01). Although exploratory, phosphoproteomics-based biomarkers deserve further investigations as predictors of afatinib efficacy.