Natural killer (NK) cells are a promising cellular therapy for cancer, with challenges in the field including persistence, functional activity, and tumor recognition. Briefly, priming blood NK cells ...with recombinant human (rh)IL-12, rhIL-15, and rhIL-18 (12/15/18) results in memory-like NK cell differentiation and enhanced responses against cancer. However, the lack of available, scalable Good Manufacturing Process (GMP)-grade reagents required to advance this approach beyond early-phase clinical trials is limiting. To address this challenge, we developed a novel platform centered upon an inert tissue factor scaffold for production of heteromeric fusion protein complexes (HFPC). The first use of this platform combined IL-12, IL-15, and IL-18 receptor engagement (HCW9201), and the second adds CD16 engagement (HCW9207). This unique HFPC expression platform was scalable with equivalent protein quality characteristics in small- and GMP-scale production. HCW9201 and HCW9207 stimulated activation and proliferation signals in NK cells, but HCW9207 had decreased IL-18 receptor signaling. RNA sequencing and multidimensional mass cytometry revealed parallels between HCW9201 and 12/15/18. HCW9201 stimulation improved NK cell metabolic fitness and resulted in the DNA methylation remodeling characteristic of memory-like differentiation. HCW9201 and 12/15/18 primed similar increases in short-term and memory-like NK cell cytotoxicity and IFNγ production against leukemia targets, as well as equivalent control of leukemia in NSG mice. Thus, HFPCs represent a protein engineering approach that solves many problems associated with multisignal receptor engagement on immune cells, and HCW9201-primed NK cells can be advanced as an ideal approach for clinical GMP-grade memory-like NK cell production for cancer therapy.
Natural killer (NK) cells are cytotoxic innate lymphoid cells that are emerging as a cellular immunotherapy for various malignancies. NK cells are particularly dependent on interleukin (IL)-15 for ...their survival, proliferation, and cytotoxic function. NK cells differentiate into memory-like cells with enhanced effector function after a brief activation with IL-12, IL-15, and IL-18. N-803 is an IL-15 superagonist composed of an IL-15 mutant (IL-15N72D) bound to the sushi domain of IL-15Rα fused to the Fc region of IgG1, which results in physiological trans-presentation of IL-15. Here, we describe the creation of a novel triple-cytokine fusion molecule, 18/12/TxM, using the N-803 scaffold fused to IL-18 via the IL-15N72D domain and linked to a heteromeric single-chain IL-12 p70 by the sushi domain of the IL-15Rα. This molecule displays trispecific cytokine activity through its binding and signaling through the individual cytokine receptors. Compared with activation with the individual cytokines, 18/12/TxM induces similar short-term activation and memory-like differentiation of NK cells on both the transcriptional and protein level and identical in vitro and in vivo anti-tumor activity. Thus, N-803 can be modified as a functional scaffold for the creation of cytokine immunotherapies with multiple receptor specificities to activate NK cells for adoptive cellular therapy.
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Fehniger and colleagues describe the creation of a novel triple-cytokine fusion molecule, 18/12/TxM, containing an interleukin (IL)-15 superagonist backbone (N-803) fused to IL-18 and IL-12. This trimeric molecule retained specific and unique IL-12, IL-15, and IL-18 activities and generated potent human memory-like natural killer cells in vitro and in vivo.
The emergence of paediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS) led to the widespread use of anti-inflammatory treatments in the absence of evidence from ...randomised controlled trials (RCTs). We aimed to assess the effectiveness of intravenous methylprednisolone compared with intravenous immunoglobulins.
This is an open-label, multicentre, two-arm RCT done at ten hospitals in Switzerland in children younger than 18 years hospitalised with PIMS-TS (defined as age <18 years; fever and biochemical evidence of inflammation, and single or multiorgan dysfunction; microbiologically proven or putative contact with SARS-CoV-2; and exclusion of any other probable disease). Patients were randomly assigned 1:1 to intravenous methylprednisolone (10 mg/kg per day for 3 days) or intravenous immunoglobulins (2 g/kg as a single dose). The primary outcome was length of hospital stay censored at day 28, death, or discharge. Secondary outcomes included proportion and duration of organ support. Analyses were done by intention-to-treat. The study was registered with Swiss National Clinical Trials Portal (SNCTP000004720) and ClinicalTrials.gov (NCT04826588).
Between May 21, 2021, and April 15, 2022, 75 patients with a median age of 9·1 years (IQR 6·2-12·2) were included in the intention-to-treat population (37 in the methylprednisolone group and 38 in the intravenous immunoglobulins group). The median length of hospital stay was 6·0 days (IQR 4·0-8·0) in the methylprednisolone group and 6·0 days (IQR 5·0-8·8) in the intravenous immunoglobulins group (estimated effect size -0·037 of the log
transformed times, 95% CI -0·13 to 0·065, p=0·42). Fewer patients in the methylprednisolone group (ten 27% of 37) required respiratory support compared with the intravenous immunoglobulin group (21 55% of 38, p=0·025). Need and duration of inotropes, admission to intensive care units, cardiac events after baseline, and major bleeding and thrombotic events were not significantly different between the study groups.
In this RCT, treatment with methylprednisolone in children with PIMS-TS did not significantly affect the length of hospital stay compared with intravenous immunoglobulins. Intravenous methylprednisolone could be an acceptable first-line treatment in children with PIMS-TS.
NOMIS Foundation, Vontobel Foundation, and Gaydoul Foundation.
Larvae of the New World gracillariid moth genus Marmara are primarily stem/bark miners, with some species mining in leaves or fruits. We describe a new species, M. viburnella Eiseman & Davis, which ...feeds on Viburnum, initially mining the leaves but completing development as a stem miner. The type series is from Nantucket Island, Massachusetts, with observations of leaf mines indicating the species is widespread in the eastern USA. Combining previously published data, our own observations, and other sources, we present a list of known Marmara hostplants, many of which represent undescribed species.
The malignant Hodgkin and Reed Sternberg (HRS) cells of classical Hodgkin lymphoma (cHL) are scarce in affected lymph nodes, creating a challenge to detect driver somatic mutations. As an alternative ...to cell purification techniques, we hypothesized that ultra-deep exome sequencing would allow genomic study of HRS cells, thereby streamlining analysis and avoiding technical pitfalls. To test this, 31 cHL tumor/normal pairs were exome sequenced to approximately 1,000× median depth of coverage. An orthogonal error-corrected sequencing approach verified >95% of the discovered mutations. We identified mutations in genes novel to cHL including: CDH5 and PCDH7, novel stop gain mutations in IL4R, and a novel pattern of recurrent mutations in pathways regulating Hippo signaling. As a further application of our exome sequencing, we attempted to identify expressed somatic single-nucleotide variants (SNV) in single-nuclei RNA sequencing (snRNA-seq) data generated from a patient in our cohort. Our snRNA analysis identified a clear cluster of cells containing a somatic SNV identified in our deep exome data. This cluster has differentially expressed genes that are consistent with genes known to be dysregulated in HRS cells (e.g., PIM1 and PIM3). The cluster also contains cells with an expanded B-cell clonotype further supporting a malignant phenotype. This study provides proof-of-principle that ultra-deep exome sequencing can be utilized to identify recurrent mutations in HRS cells and demonstrates the feasibility of snRNA-seq in the context of cHL. These studies provide the foundation for the further analysis of genomic variants in large cohorts of patients with cHL.
Our data demonstrate the utility of ultra-deep exome sequencing in uncovering somatic variants in Hodgkin lymphoma, creating new opportunities to define the genes that are recurrently mutated in this disease. We also show for the first time the successful application of snRNA-seq in Hodgkin lymphoma and describe the expression profile of a putative cluster of HRS cells in a single patient.
Evaluate the cost–effectiveness of ocriplasmin in symptomatic vitreomacular adhesion (VMA) with or without full-thickness macular hole ≤400 μm versus standard of care.
A state-transition model ...simulated a cohort through disease health states; assignment of utilities to health states reflected the distribution of visual acuity. Efficacy of ocriplasmin was derived from logistic regression models using Ocriplasmin for Treatment for Symptomatic Vitreomacular Adhesion Including Macular Hole trial data. Model inputs were extracted from Phase III trials and published literature. The analysis was conducted from a US Medicare perspective.
Lifetime incremental cost–effectiveness ratio was US$4887 per quality-adjusted life year gained in the total population, US$4255 and US$10,167 in VMA subgroups without and with full-thickness macular hole, respectively.
Ocriplasmin was cost effective compared with standard of care in symptomatic VMA.
The purpose of this study was to determine whether walking is associated with a reduced risk of cardiovascular disease hospitalization and death in community-dwelling older men and women.
A ...prospective study, with follow-up time of 4 to 5 years (average 4.2 years).
A western Washington health maintenance organization.
Men and women aged 65 years and older from a random sample of HMO enrollees invited by mail to participate in a health promotion intervention trial (36% accepted the invitation and completed questionnaires). This report is based on 1645 older adults without severe disability and without history of heart disease. Vital status ascertainment was complete (100%), and only 2.6% did not complete the follow-up.
Reported frequency and duration of walking for exercise, work, errands, pleasure, and hiking in the 2 weeks before baseline were used to classify hours of walking per week. The two main outcomes were: (1) cardiovascular disease hospitalizations with a discharge diagnosis of coronary (ICD-9-CM 410-414) or other cardiovascular diseases (ICD-9-CM 390-409, 415-448) documented by computerized hospitalization records and (2) death. Numerous potential confounding factors were considered, including age, sex, treated high blood pressure, current estrogen use and chronic disease score (ascertained by computerized medical and pharmacy records), and ethnicity, education, income, physical function, self-rated health status, smoking, alcohol intake, and body mass index (ascertained by self-report on the mailed questionnaire).
Walking more than 4 hours/week was associated significantly with a reduced risk of cardiovascular disease hospitalization in both sexes combined compared with walking less than 1 hour/week (age and sex-adjusted relative risk = 0.69; 95% confidence interval, 0.52-0.90). This association was not altered by adjustment for baseline cardiovascular risk factors and indicators of general health status. The association was present in all age groups, among those with and without physical limitations, and also among those who did and did not also participate in more vigorous physical activities. Walking more than 4 hours/week was also associated with a reduced risk of death (age and sex-adjusted relative risk = 0.73; 95% confidence interval, 0.48-1.10), however, this association was substantially diminished by adjustment for cardiovascular risk factors and measures of general health status.
Walking more than 4 hours/week may reduce the risk of hospitalization for cardiovascular disease events. The association of walking more than 4 hours/week with reduced risk of death may be mediated by effects of walking on other risk factors. These findings provide much stronger evidence than previously available for advising older men and women to embark on or maintain a sustained program of walking to prevent cardiovascular disease events.
Deep learning (DL) can accelerate the prediction of prognostic biomarkers from routine pathology slides in colorectal cancer (CRC). However, current approaches rely on convolutional neural networks ...(CNNs) and have mostly been validated on small patient cohorts. Here, we develop a new transformer-based pipeline for end-to-end biomarker prediction from pathology slides by combining a pre-trained transformer encoder with a transformer network for patch aggregation. Our transformer-based approach substantially improves the performance, generalizability, data efficiency, and interpretability as compared with current state-of-the-art algorithms. After training and evaluating on a large multicenter cohort of over 13,000 patients from 16 colorectal cancer cohorts, we achieve a sensitivity of 0.99 with a negative predictive value of over 0.99 for prediction of microsatellite instability (MSI) on surgical resection specimens. We demonstrate that resection specimen-only training reaches clinical-grade performance on endoscopic biopsy tissue, solving a long-standing diagnostic problem.
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•AI-based prediction of biomarkers (MSI, BRAF, and KRAS) using transformers•MSI prediction reaches clinical-grade performance on biopsies of colorectal cancer•Transformer-based biomarker prediction generalizes better and is more data efficient•Large-scale multi-cohort evaluation on over 13,000 patients from 16 cohorts
Wagner et al. show that transformer-based prediction of biomarkers from histology substantially improves the performance, generalizability, data efficiency, and interpretability as compared with current state-of-the-art algorithms. The method significantly outperforms existing approaches for microsatellite instability detection in surgical resections and reaches clinical-grade performance on biopsies of colorectal cancer, solving a long-standing diagnostic problem.
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