SARS-CoV-2 recombinant spike ferritin nanoparticle vaccine adjuvanted with Army Liposome Formulation containing monophosphoryl lipid A and QS-21: a phase 1, randomised, double-blind, placebo-controlled, first-in-human clinical trial.

BACKGROUND: A self-assembling SARS-CoV-2 WA-1 recombinant spike ferritin nanoparticle (SpFN) vaccine co-formulated with Army Liposomal Formulation (ALFQ) adjuvant containing monophosphoryl lipid A and QS-21 (SpFN/ALFQ) has shown protective efficacy in animal challenge models. This trial aims to assess the safety and immunogenicity of SpFN/ALFQ in a first-in-human clinical trial. METHODS: In this phase 1, randomised, double-blind, placebo-controlled, first-in-human clinical trial, adults were randomly assigned (5:5:2) to receive 25 µg or 50 µg of SpFN/ALFQ or saline placebo intramuscularly at day 1 and day 29, with an optional open-label third vaccination at day 181. Enrolment and randomisation occurred sequentially by group; randomisation was done by an interactive web-based randomisation system and only designated unmasked study personnel had access to the randomisation code. Adults were required to be seronegative and unvaccinated for inclusion. Local and systemic reactogenicity, adverse events, binding and neutralising antibodies, and antigen-specific T-cell responses were quantified. For safety analyses, exact 95% Clopper-Pearson CIs for the probability of any incidence of an unsolicited adverse event was computed for each group. For immunogenicity results, CIs for binary variables were computed using the exact Clopper-Pearson methodology, while CIs for geometric mean titres were based on 10 000 empirical bootstrap samples. Post-hoc, paired one-sample t tests were used to assess the increase in mean log-10 neutralising antibody titres between day 29 and day 43 (after the second vaccination) for the primary SARS-CoV-2 targets of interest. This trial is registered at ClinicalTrials.gov, NCT04784767, and is closed to new participants. FINDINGS: Between April 7, and June 29, 2021, 29 participants were enrolled in the study. 20 individuals were assigned to receive 25 µg SpFN/ALFQ, four to 50 µg SpFN/ALFQ, and five to placebo. Neutralising antibody responses peaked at day 43, 2 weeks after the second dose. Neutralisation activity against multiple omicron subvariants decayed more slowly than against the D614G or beta variants until 5 months after second vaccination for both dose groups. CD4+ T-cell responses were elicited 4 weeks after the first dose and were boosted after a second dose of SpFN/ALFQ for both dose groups. Neutralising antibody titres against early omicron subvariants and clade 1 sarbecoviruses were detectable after two immunisations and peaked after the third immunisation for both dose groups. Neutralising antibody titres against XBB.1.5 were detected after three vaccinations. Passive IgG transfer from vaccinated volunteers into Syrian golden hamsters controlled replication of SARS-CoV-1 after challenge. INTERPRETATION: SpFN/ALFQ was well tolerated and elicited robust and durable binding antibody and neutralising antibody titres against a broad panel of SARS-CoV-2 variants and other sarbecoviruses. FUNDING: US Department of Defense, Defense Health Agency.

Implementing good life with osteoArthritis from Denmark (GLA:D®) in Australian public hospitals. Part 2: Service providers' perceptions.

BACKGROUND: Previous literature has reported the successful implementation of the Good Life with osteoArthritis in Denmark (GLA:D®) program into predominantly private practice settings. There may be unique challenges present within the public hospital setting that influence GLA:D® implementation in public health. OBJECTIVE: Explore the attitudes and experiences of service providers directly involved in implementing GLA:D® in Australian public tertiary hospitals. DESIGN: Qualitative descriptive study design. METHOD: Service providers (n = 14) from three participating hospitals took part in semi-structured focus groups at the completion of the 6-month implementation period. Inductive thematic analysis was employed to identify primary domains across all facilities. RESULTS: Four broad domains were identified. Factors that influenced uptake included GLA:D® being a recognisable, evidence-based product requiring minimal development or adaptation. The fidelity of the GLA:D® Australia program was challenged by referral of patients with multiple/complex medical comorbidities, and patient preference to complete registry data via paper rather than online. Several operational considerations are required when delivering GLA:D® in a public hospital setting, including adequate numbers of GLA:D®-trained staff, additional screening requirements, obtaining appropriate clinical space, and persisting patient barriers to attending the service. GLA:D® provided benefits beyond improvement in pain and function, including social interactivity, high attendance and promotion of long-term self-management, while also maximising service efficiencies. CONCLUSIONS: Implementing GLA:D® in Australian public hospitals was supported by service providers. Specific operational and administrative factors, including staff training, patient complexity, and registry requirements should be considered when attempting to embed and sustain GLA:D® in large Australian public tertiary hospitals.

Implementing good life with osteoArthritis from Denmark (GLA:D®) in australian public hospitals. Part 1: Feasibility.

BACKGROUND: Literature reporting positive outcomes from the Good Life with osteoArthritis in Denmark (GLA:D®) program in Australia mainly involves patients attending private physiotherapy services. OBJECTIVE: Evaluate the feasibility of implementing GLA:D® in Australian public hospitals. DESIGN: Implementation study in three metropolitan tertiary public hospitals over six months. METHOD: Patients aged ≥18 years with knee or hip joint-related problems deemed appropriate for non-surgical care were invited to participate in GLA:D®. Feasibility was evaluated using RE-AIM framework components (Implementation, Effectiveness, Maintenance) using service-level metrics, patient-level data, and program fidelity assessment. Findings of qualitative interviews with service providers are presented in Part 2. RESULTS: Implementation: 70 patients (69 with knee osteoarthritis) participated (13 cohorts). 55 (79%) patients attended both education sessions, and 49 patients (70%) attended 10-12 exercises sessions. Fidelity was met based on environmental, therapist, participant- and program-related criteria. EFFECTIVENESS: At 3 months, patients reported lower average pain (visual analogue scale [0-100 mm]: effect size -0.56, 95% CI -0.88 to -0.23) and disability (HOOS/KOOS-12 [100-0]: 0.67, 0.28 to 1.05), and improved quality of life (EQ-5D overall score: 0.46, 0.11 to 0.80). No adverse events were reported. All patients who completed 3-month assessment (n = 52) would recommend GLA:D®. Maintenance: All participating services elected to continue delivering GLA:D® beyond the study. CONCLUSIONS: Implementing GLA:D® in Australian public hospitals is feasible, safe, and acceptable to patients with knee osteoarthritis. Public hospital patients with knee osteoarthritis reported improvements in pain, disability, and quality of life similar to previous GLA:D® cohorts.

Off-label prescribing of immune checkpoint inhibitor therapy at a single pediatric cancer center.

BACKGROUND: Immune checkpoint inhibitors (ICI) have improved outcomes in a variety of adult cancers and are prescribed with increasing frequency across oncology. However, patterns of off-label use of ICI in pediatrics remain unclear. METHODS: This is a single-institution, retrospective cohort study evaluating off-label ICI use in pediatric and young adult patients with cancer treated at our institution from 2014 to 2022. Response was based on clinician assessment derived from clinical records. Immune-related adverse events (iRAEs) were classified according to CTCAE v5.0. RESULTS: We identified 50 unique patients treated with off-label ICI (28 with solid tumors, 20 with central nervous system (CNS) tumors, 2 with hematologic malignancies). At time of ICI initiation, only five patients (10%) had localized disease, and all but one patient was treated in the relapsed/refractory setting. All patients were treated with the FDA-approved weight-based dosing recommendations. Overall, there was disease control in 21 patients (42%), with best response including one complete response (melanoma), two partial responses (high-grade glioma, CNS nongerminomatous germ cell tumor), and 18 patients with stable disease. Forty-four patients (88%) eventually experienced disease progression. Among 22 patients (44%) experiencing iRAEs, 10 (20%) had a grade ≥3 irAE, 12 (24%) required corticosteroids, and 14 (28%) required ICI discontinuation. irAE occurrence was associated with significantly improved progression-free survival (HR 0.35; 95% CI: 0.18 to 0.68; p = 0.002) and overall survival (HR 0.33; 95% CI: 0.17 to 0.66; p = 0.002). CONCLUSIONS: At our institution, ICI was most commonly prescribed in the relapsed/refractory setting to patients with metastatic disease. The treatment was generally well-tolerated in the pediatric population. The overall response rate was low, and the majority of patients eventually experienced disease progression. A few patients, however, had durable treatment responses. Further studies are needed to identify which pediatric patients are most likely to benefit from ICI.

An exploration of footwear preferences, attitudes and beliefs in people with knee osteoarthritis: A qualitative study.

BACKGROUND: Although footwear can improve pain and function in individuals with knee osteoarthritis (OA), perspectives about footwear in this population have not been explored. OBJECTIVES: This qualitative study explored preferences, attitudes and beliefs about footwear in adults with knee OA. METHODS: Twenty individuals with a clinical diagnosis of knee OA (aged 45-79 years, 65% women) participated in semi-structured interviews about factors which influence footwear selection, the effect of footwear on knee symptoms, and footwear modifications. Data were analysed thematically. RESULTS: Four themes, with sub-themes, were identified: i) there are specific footwear characteristics people look for, with comfort as their top priority; ii) shoe appearance is important; iii) footwear can aggravate or ease symptoms; and iv) people with knee OA find footwear in a variety of ways. Participants related built-in arch support, a cushioned insole and low/no heel, without addition of foot orthoses, to comfort, and were willing to pay more for comfort and quality. Appearance was also a consideration, and participants indicated they would tolerate short periods of symptom aggravation for aesthetic shoes. Participants felt that footwear choice affected their knee symptoms and risk of slipping/twisting. Participants reported that their footwear choices were determined through trial-and-error, and sometimes on advice from health professionals or shoe store salespersons. CONCLUSIONS: There are specific footwear features important to individuals with knee OA. Knowledge of these features can be used by health professionals to inform footwear discussions with knee OA patients and serve as considerations when developing footwear targeted for this population.

A spatial cell atlas of neuroblastoma reveals developmental, epigenetic and spatial axis of tumor heterogeneity.

Neuroblastoma is a pediatric cancer arising from the developing sympathoadrenal lineage with complex inter- and intra-tumoral heterogeneity. To chart this complexity, we generated a comprehensive cell atlas of 55 neuroblastoma patient tumors, collected from two pediatric cancer institutions, spanning a range of clinical, genetic, and histologic features. Our atlas combines single-cell/nucleus RNA-seq (sc/scRNA-seq), bulk RNA-seq, whole exome sequencing, DNA methylation profiling, spatial transcriptomics, and two spatial proteomic methods. Sc/snRNA-seq revealed three malignant cell states with features of sympathoadrenal lineage development. All of the neuroblastomas had malignant cells that resembled sympathoblasts and the more differentiated adrenergic cells. A subset of tumors had malignant cells in a mesenchymal cell state with molecular features of Schwann cell precursors. DNA methylation profiles defined four groupings of patients, which differ in the degree of malignant cell heterogeneity and clinical outcomes. Using spatial proteomics, we found that neuroblastomas are spatially compartmentalized, with malignant tumor cells sequestered away from immune cells. Finally, we identify spatially restricted signaling patterns in immune cells from spatial transcriptomics. To facilitate the visualization and analysis of our atlas as a resource for further research in neuroblastoma, single cell, and spatial-omics, all data are shared through the Human Tumor Atlas Network Data Commons at www.humantumoratlas.org.

Early detection of malignant and pre-malignant peripheral nerve tumors using cell-free DNA fragmentomics.

Early detection of neurofibromatosis type 1 (NF1) associated peripheral nerve sheath tumors (PNST) informs clinical decision-making, potentially averting deadly outcomes. Here, we describe a cell-free DNA (cfDNA) fragmentomic approach which distinguishes non-malignant, pre-malignant and malignant forms of NF1 PNST. Using plasma samples from a novel cohort of 101 NF1 patients and 21 healthy controls, we validated that our previous cfDNA copy number alteration (CNA)-based approach identifies malignant peripheral nerve sheath tumor (MPNST) but cannot distinguish among benign and premalignant states. We therefore investigated the ability of fragment-based cfDNA features to differentiate NF1-associated tumors including binned genome-wide fragment length ratios, end motif analysis, and non-negative matrix factorization deconvolution of fragment lengths. Fragmentomic methods were able to differentiate pre-malignant states including atypical neurofibromas (AN). Fragmentomics also adjudicated AN cases suspicious for MPNST, correctly diagnosing samples noninvasively, which could have informed clinical management. Overall, this study pioneers the early detection of malignant and premalignant peripheral nerve sheath tumors in NF1 patients using plasma cfDNA fragmentomics. In addition to screening applications, this novel approach distinguishes atypical neurofibromas from benign plexiform neurofibromas and malignant peripheral nerve sheath tumors, enabling more precise clinical diagnosis and management.

Diverse array of neutralizing antibodies elicited upon Spike Ferritin Nanoparticle vaccination in rhesus macaques.

The repeat emergence of SARS-CoV-2 variants of concern (VoC) with decreased susceptibility to vaccine-elicited antibodies highlights the need to develop next-generation vaccine candidates that confer broad protection. Here we describe the antibody response induced by the SARS-CoV-2 Spike Ferritin Nanoparticle (SpFN) vaccine candidate adjuvanted with the Army Liposomal Formulation including QS21 (ALFQ) in non-human primates. By isolating and characterizing several monoclonal antibodies directed against the Spike Receptor Binding Domain (RBD), N-Terminal Domain (NTD), or the S2 Domain, we define the molecular recognition of vaccine-elicited cross-reactive monoclonal antibodies (mAbs) elicited by SpFN. We identify six neutralizing antibodies with broad sarbecovirus cross-reactivity that recapitulate serum polyclonal antibody responses. In particular, RBD mAb WRAIR-5001 binds to the conserved cryptic region with high affinity to sarbecovirus clades 1 and 2, including Omicron variants, while mAb WRAIR-5021 offers complete protection from B.1.617.2 (Delta) in a murine challenge study. Our data further highlight the ability of SpFN vaccination to stimulate cross-reactive B cells targeting conserved regions of the Spike with activity against SARS CoV-1 and SARS-CoV-2 variants.

Lassa virus in novel hosts: insights into the epidemiology of lassa virus infections in southern Nigeria.

Identification of the diverse animal hosts responsible for spill-over events from animals to humans is crucial for comprehending the transmission patterns of emerging infectious diseases, which pose significant public health risks. To better characterize potential animal hosts of Lassa virus (LASV), we assessed domestic and non-domestic animals from 2021-2022 in four locations in southern Nigeria with reported cases of Lassa fever (LF). Birds, lizards, and domestic mammals (dogs, pigs, cattle and goats) were screened using RT-qPCR, and whole genome sequencing was performed for lineage identification on selected LASV positive samples. Animals were also screened for exposure to LASV by enzyme-linked immunosorbent assay (ELISA). Among these animals, lizards had the highest positivity rate by PCR. Genomic sequencing of samples in most infected animals showed sub-lineage 2 g of LASV. Seropositivity was highest among cattle and lowest in pigs. Though the specific impact these additional hosts may have in the broader virus-host context are still unknown - specifically relating to pathogen diversity, evolution, and transmission - the detection of LASV in non-rodent hosts living in proximity to confirmed human LF cases suggests their involvement during transmission as potential reservoirs. Additional epidemiological data comparing viral genomes from humans and animals, as well as those circulating within the environment will be critical in understanding LASV transmission dynamics and will ultimately guide the development of countermeasures for this zoonotic health threat.

Response to letter to editor: The association between hip strength, physical function and dynamic balance in people with unilateral knee osteoarthritis: A cross-sectional study.

We thank the reader for their interest and response to our study investigating the association between clinical measures of hip strength in multiple directions and physical function (including dynamic balance) in people with knee osteoarthritis. Below, we provide a response to their questions, in turn.

Protocol for a randomised feasibility trial comparing a combined program of education and exercise versus general advice for ankle osteoarthritis.

BACKGROUND: Ankle osteoarthritis (OA) is a serious problem with high associated pain and disability. While education and exercise are recommended for the initial management of OA, this has not been investigated in ankle OA. The primary aim of this study is to establish the feasibility of running a full-scale randomised controlled trial (RCT) investigating a combined education and exercise program compared to a general advice program for people with ankle OA. The secondary aims are to collect preliminary data which will inform sample size calculations, and understand the perspectives of people with ankle OA on their participation in the trial. METHODS: Thirty individuals aged 35 years or older with symptomatic radiographic ankle OA will be recruited from the community and randomised to receive either a combined education and exercise program or a general advice program, both of which will be delivered by a physiotherapist in a group setting. Primary outcomes of feasibility include responses to study advertisements, number of eligible participants, recruitment rate, adherence with the intervention, fidelity of the intervention, adverse events, drop-out rate, and credibility and expectancy of the intervention. Secondary participant-reported outcomes will include global rating of change, patient acceptable symptom state, severity of ankle pain and stiffness, self-reported function, quality of life, satisfaction with treatment, and use of co-interventions. Follow up will be at 8 weeks and 3 months. Physical measures of 40 m walking speed, timed stairs descent, heel raise endurance and ankle dorsiflexion range of motion will be collected at baseline and 8 weeks. Primary feasibility outcomes will be reported descriptively, and estimates of the variability of secondary participant-reported and physical outcomes will be calculated. Semi-structured interviews will be conducted with participants to understand perspectives about the intervention and participation in the trial, with data analyzed thematically. DISCUSSION: Study findings will establish the feasibility of running a full-scale RCT to investigate a combined education and exercise program compared to a general advice program for people with ankle OA. This study is a necessary first step to advance the international research agenda of evaluating the efficacy of exercise in the management of ankle OA. TRIAL REGISTRATION: ACTRN12623000017628. Registered 10 January 2023, https://www.anzctr.org.au/ACTRN12623000017628.aspx .

Effects of foot orthoses and footwear interventions on impairments and quality of life in people with hip pain: A systematic review.

BACKGROUND: Foot orthoses and footwear interventions are advocated for the management of lower limb musculoskeletal conditions including the hip, but much of the research is focused on knee disorders. The aim of this systematic review was to synthesise the literature that investigates the use of foot orthoses or footwear in people with hip-related pain. METHODS: MEDLINE, EMBASE, CINAHL, AMED and SPORTDiscus were searched from inception to March 2023. Randomised controlled trials (RCT), cohort and pre-post studies reporting on footwear and foot orthoses interventions, in participants with hip-related pain, were eligible for inclusion. Outcomes included pain, physical function, and quality of life (QoL). Effect sizes were calculated where sufficient data were available. Reporting quality was assessed using the Cochrane Risk of Bias Tool (Rob-2) and the Joanna Briggs Institute Checklist. The overall quality of evidence was rated according to the Grading of Recommendations, Assessment, Development, and Evaluations framework. RESULTS: Of the seven included studies (n = 266 participants), there was one RCT, one cohort and five single-group pre-post designs. Interventions included customised and non-customised arch supports, heel lifts, and footwear modifications, used in the following hip conditions: trochanteric pain, non-specific hip pain, hip osteoarthritis, and leg length dysfunction following total hip arthroplasty. Meta-analysis was possible for outcomes in two studies, demonstrating moderate improvement in pain following foot orthoses use. Overall certainty of evidence ranged from very low to low. CONCLUSION: Single-group pre-post study designs describe positive relationships between foot orthoses and footwear use and improvements in hip pain, function, and QoL. However, these results were not supported by the only available RCT. Given this is a relatively inexpensive and non-invasive treatment approach, further rigorous studies are warranted.

A recombinant virus-like particle vaccine against adenovirus-7 induces a potent humoral response.

Adenoviruses (AdVs) cause infections in humans that range from mild to severe, and can cause outbreaks particularly in close contact settings. Several human AdV types have been identified, which can cause a wide array of clinical manifestations. AdV types 4 and 7 (AdV-4 and AdV-7), which are among the most commonly circulating types in the United States, are known to cause acute respiratory disease that can result in hospitalization and rarely, death. Currently, the only vaccines approved for use in humans are live virus vaccines against AdV-4 and AdV-7, though these vaccines are only authorized for use in U.S. military personnel. While they are efficacious, use of these live virus vaccines carries considerable risks of vaccine-associated viral shedding and recombination. Here, we present an alternative vaccination strategy against AdV-7 using the virus-like particle platform (AdVLP-7). We describe the production of stable recombinant AdVLP-7, and demonstrate that AdVLP-7 is structurally analogous to wild-type AdV-7 virions (WT AdV-7). Preclinical immunogenicity studies in mice show that AdVLP-7 elicits a potent humoral immune response, comparable to that observed in mice immunized with WT AdV-7. Specifically, AdVLP-7 induces high titers of antibodies against AdV-7-specific antigens that can effectively neutralize AdV-7.

Differential Genome Replication of a Unique Single-Amino-Acid Mutation in the Adenovirus-4 Component of the Live Oral Adenovirus Type 4 and Type 7 Vaccine.

The FDA-approved Adenovirus Type 4 and Type 7 Vaccine, Live, Oral is highly effective and essential in preventing acute respiratory diseases (ARDs) in U.S. military recruits. Our study revealed the presence of a previously undetected mutation, not found in the wild-type human adenovirus type 4 (HAdV-4) component of the licensed vaccine, which contains an amino acid substitution (P388T) in the pre-terminal protein (pTP). This study demonstrated that replication of the T388 HAdV-4 vaccine mutant virus is favored over the wild type in WI-38 cells, the cell type utilized in vaccine manufacturing. However, results from serial human stool specimens of vaccine recipients support differential genome replication in the gastrointestinal tract (GI), demonstrated by the steady decline of the percentage of mutant T388 vaccine virus. Since vaccine efficacy depends upon GI replication and the subsequent immune response, the mutation can potentially impact vaccine efficacy.

Neuromechanical Properties of the Vastus Medialis and Vastus Lateralis in Adolescents With Patellofemoral Pain.

Background: An alteration in the force distribution among quadriceps heads is one possible underlying mechanism of patellofemoral pain. However, this hypothesis cannot be directly tested as there are currently no noninvasive experimental techniques to measure individual muscle force or torque in vivo in humans. In this study, the authors considered a combination of biomechanical and muscle activation measures, which enabled us to estimate the mechanical impact of the vastus medialis (VM) and vastus lateralis (VL) on the patella. Purpose/Hypothesis: The purpose of this study was to determine whether the relative index of torque distribution for the VM and VL differs between adolescents with and without patellofemoral pain. It was hypothesized that, relative to the VL, the VM would contribute less to knee extension torque in adolescents with patellofemoral pain compared with controls. Study Design: Cross-sectional study; Level of evidence, 3. Methods: Twenty adolescents with patellofemoral pain and 20 matched control participants were included (38 female; age, 15.3 ± 1.8 years; weight, 58 ± 13 kg; height, 164 ± 8 cm). Muscle volumes and resting moment arms were quantified from magnetic resonance images, and fascicle lengths were obtained from panoramic B-mode ultrasonography. Muscle activation was estimated using surface electromyography during submaximal isometric tasks (wall-squat and seated tasks). Muscle torque was estimated as the product of muscle physiological cross-sectional area (ie, muscle volume/fascicle length), muscle activation (normalized to maximal activation), and moment arm. Results: Across tasks and force levels, the relative contribution of the VM to the overall medial and lateral vastii torque was 31.0% ± 8.6% for controls and 31.5 ± 7.6% for adolescents with patellofemoral pain (group effect, P > .34). Conclusion: For the tasks and positions investigated in this study, the authors found no evidence of lower VM torque generation (relative to the VL) in adolescents with patellofemoral pain compared with controls.

A Phase 1 Two-Arm, Randomized, Double-Blind, Active-Controlled Study of Live, Oral Plasmid-Derived Adenovirus Type 4 and Type 7 Vaccines in Seronegative Adults.

PXVX0047 is an investigational vaccine developed for active immunization to prevent febrile acute respiratory disease (ARD) caused by adenovirus serotypes 4 (Ad4) and 7 (Ad7). PXVX0047 consists of a modernized, plasmid-derived vaccine that was generated using a virus isolated from Wyeth Ad4 and Ad7 vaccine tablets. A phase 1 two-arm, randomized, double-blind, active-controlled study was conducted to evaluate the safety profile and immunogenicity of the investigational adenovirus vaccines. The two components of PXVX0047 were administered orally together in a single dose to 11 subjects. For comparison, three additional subjects received the Ad4/Ad7 vaccine that is currently in use by the US military. The results of this study show that the tolerability and immunogenicity of the PXVX0047 Ad7 component are comparable with that of the control Ad4/Ad7 vaccine; however, the immunogenicity of the PXVX0047 Ad4 component was lower than expected. Clinical trial number NCT03160339.

Efficacy of Nivolumab in Pediatric Cancers with High Mutation Burden and Mismatch Repair Deficiency.

PURPOSE: Checkpoint inhibitors have limited efficacy for children with unselected solid and brain tumors. We report the first prospective pediatric trial (NCT02992964) using nivolumab exclusively for refractory nonhematologic cancers harboring tumor mutation burden (TMB) ≥5 mutations/megabase (mut/Mb) and/or mismatch repair deficiency (MMRD). PATIENTS AND METHODS: Twenty patients were screened, and 10 were ultimately included in the response cohort of whom nine had TMB >10 mut/Mb (three initially eligible based on MMRD) and one patient had TMB between 5 and 10 mut/Mb. RESULTS: Delayed immune responses contributed to best overall response of 50%, improving on initial objective responses (20%) and leading to 2-year overall survival (OS) of 50% [95% confidence interval (CI), 27-93]. Four children, including three with refractory malignant gliomas are in complete remission at a median follow-up of 37 months (range, 32.4-60), culminating in 2-year OS of 43% (95% CI, 18.2-100). Biomarker analyses confirmed benefit in children with germline MMRD, microsatellite instability, higher activated and lower regulatory circulating T cells. Stochastic mutation accumulation driven by underlying germline MMRD impacted the tumor microenvironment, contributing to delayed responses. No benefit was observed in the single patient with an MMR-proficient tumor and TMB 7.4 mut/Mb. CONCLUSIONS: Nivolumab resulted in durable responses and prolonged survival for the first time in a pediatric trial of refractory hypermutated cancers including malignant gliomas. Novel biomarkers identified here need to be translated rapidly to clinical care to identify children who can benefit from checkpoint inhibitors, including upfront management of cancer. See related commentary by Mardis, p. 4701.

Sex differences in the transcriptome of extracellular vesicles secreted by fetal neural stem cells and effects of chronic alcohol exposure.

BACKGROUND: Prenatal alcohol (ethanol) exposure (PAE) results in brain growth restriction, in part, by reprogramming self-renewal and maturation of fetal neural stem cells (NSCs) during neurogenesis. We recently showed that ethanol resulted in enrichment of both proteins and pro-maturation microRNAs in sub-200-nm-sized extracellular vesicles (EVs) secreted by fetal NSCs. Moreover, EVs secreted by ethanol-exposed NSCs exhibited diminished efficacy in controlling NSC metabolism and maturation. Here we tested the hypothesis that ethanol may also influence the packaging of RNAs into EVs from cell-of-origin NSCs. METHODS: Sex-specified fetal murine iso-cortical neuroepithelia from three separate pregnancies were maintained ex vivo, as neurosphere cultures to model the early neurogenic niche. EVs were isolated by ultracentrifugation from NSCs exposed to a dose range of ethanol. RNA from paired EV and cell-of-origin NSC samples was processed for ribosomal RNA-depleted RNA sequencing. Differential expression analysis and exploratory weighted gene co-expression network analysis (WGCNA) identified candidate genes and gene networks that were drivers of alterations to the transcriptome of EVs relative to cells. RESULTS: The RNA content of EVs differed significantly from cell-of-origin NSCs. Biological sex contributed to unique transcriptome variance in EV samples, where > 75% of the most variant transcripts were also sex-variant in EVs but not in cell-of-origin NSCs. WGCNA analysis also identified sex-dependent enrichment of pathways, including dopamine receptor binding and ectoderm formation in female EVs and cell-substrate adhesion in male EVs, with the top significant DEGs from differential analysis of overall individual gene expressions, i.e., Arhgap15, enriched in female EVs, and Cenpa, enriched in male EVs, also serving as WCGNA hub genes of sex-biased EV WGCNA clusters. In addition to the baseline RNA content differences, ethanol exposure resulted in a significant dose-dependent change in transcript expression in both EVs and cell-of-origin NSCs that predominantly altered sex-invariant RNAs. Moreover, at the highest dose, ~ 73% of significantly altered RNAs were enriched in EVs, but depleted in NSCs. CONCLUSIONS: The EV transcriptome is distinctly different from, and more sex-variant than, the transcriptome of cell-of-origin NSCs. Ethanol, a common teratogen, results in dose-dependent sorting of RNA transcripts from NSCs to EVs which may reprogram the EV-mediated endocrine environment during neurogenesis.

Automating Case Reporting of Chlamydia and Gonorrhea to Public Health Authorities in Illinois Clinics: Implementation and Evaluation of Findings.

BACKGROUND: Chlamydia and gonorrhea cases continue to rise in Illinois, increasing by 16.4% and 70.9% in 2019, respectively, compared with 2015. Providers are required to report both chlamydia and gonorrhea, as mandated by public health laws. Manual reporting remains a huge burden; 90%-93% of cases were reported to Illinois Department of Public Health (IDPH) via electronic laboratory reporting (ELR), and the remaining were reported through web-based data entry platforms, faxes, and phone calls. However, cases reported via ELRs only contain information available to a laboratory facility and do not contain additional data needed for public health. Such data are typically found in an electronic health record (EHR). Electronic case reports (eCRs) were developed and automated the generation of case reports from EHRs to be reported to public health agencies. OBJECTIVE: Prior studies consolidated trigger criteria for eCRs, and compared with manual reporting, found it to be more complete. The goal of this project is to pilot standards-based eCR for chlamydia and gonorrhea. We evaluated the throughput, completeness, and timeliness of eCR compared to ELR, as well as the implementation experience at a large health center-controlled network in Illinois. METHODS: For this study, we selected 8 clinics located on the north, west, and south sides of Chicago to implement the eCRs; these cases were reported to IDPH. The study period was 52 days. The centralized EHR used by these clinics leveraged 2 of the 3 case detection scenarios, which were previously defined as the trigger, to generate an eCR. These messages were successfully transmitted via Health Level 7 electronic initial case report standard. Upon receipt by IDPH, these eCRs were parsed and housed in a staging database. RESULTS: During the study period, 183 eCRs representing 135 unique patients were received by IDPH. eCR reported 95% (n=113 cases) of all the chlamydia cases and 97% (n=70 cases) of all the gonorrhea cases reported from the participating clinical sites. eCR found an additional 14 (19%) cases of gonorrhea that were not reported via ELR. However, ELR reported an additional 6 cases of chlamydia and 2 cases of gonorrhea, which were not reported via eCR. ELR reported 100% of chlamydia cases but only 81% of gonorrhea cases. While key elements such as patient and provider names were complete in both eCR and ELR, eCR was found to report additional clinical data, including history of present illness, reason for visit, symptoms, diagnosis, and medications. CONCLUSIONS: eCR successfully identified and created automated reports for chlamydia and gonorrhea cases in the implementing clinics in Illinois. eCR demonstrated a more complete case report and represents a promising future of reducing provider burden for reporting cases while achieving greater semantic interoperability between health care systems and public health.

Clustered cases of human adenovirus types 4, 7, and 14 infections in US Department of Defense Beneficiaries during the 2018-2019 season.

Human adenoviruses (HAdV) are genetically diverse and can infect a number of tissues with severities varied from mild to fatal. HAdV types 3, 4, 7, 11, 14, 21, and 55 were associated with acute respiratory illnesses outbreaks in the United States and in other countries. The risk of outbreaks can be effectively controlled by HAdV vaccination or mitigated by screening and preventive measures. During the influenza season 2018-2019, the DoD Global Respiratory Pathogen Surveillance Program (DoDGRS) received 24 300 respiratory specimens. HAdV samples that produced positive cytopathic effects in viral cultivation were subjected to next-generation sequencing for genome sequence assembly, genome typing, whole genome phylogeny, and sequence comparative analyses. A variety of HAdV types were identified in this study, including HAdV types 1-7, 14, 55, and 56. HAdV types 4, 7, and 14 were found in clustered cases in Colorado, Florida, New York, and South Carolina. Comparative sequence analyses of these isolates revealed the emergence of novel genetic mutations despite the stability of adenovirus genomes. Genomic surveillance of HAdV suggested possible undetected outbreaks and shed light on prevalence, genetic divergence, and viral evolution of HAdV. Continued surveillance will inform risk assessment and countermeasures.