Additional Signal Research Articles

RSVP Keyboard with Inquiry Preview: mixed performance and user experience with an adaptive, multimodal typing interface combining EEG and switch input.

The RSVP Keyboard is a non-implantable, event-related potential-based brain-computer interface (BCI) system designed to support communication access for people with severe speech and physical impairments. Here we introduce Inquiry Preview, a new RSVP Keyboard interface incorporating switch input for users with some voluntary motor function, and describe its effects on typing performance and other outcomes. Four individuals with disabilities participated in the collaborative design of possible switch input applications for the RSVP Keyboard, leading to the development of Inquiry Preview and a method of fusing switch input with language model and electroencephalography (EEG) evidence for typing. Twenty-four participants without disabilities and one potential end user with incomplete locked-in syndrome took part in two experiments investigating the effects of Inquiry Preview and two modes of switch input on typing accuracy and speed during a copy-spelling task. For participants without disabilities, Inquiry Preview and switch input tended to worsen typing performance compared to the standard RSVP Keyboard condition, with more consistent effects across participants for speed than for accuracy. However, there was considerable variability, with some participants demonstrating improved typing performance and better user experience with Inquiry Preview and switch input. Typing performance for the potential end user was comparable to that of participants without disabilities. He typed most quickly and accurately with Inquiry Preview and switch input and gave favorable user experience ratings to those conditions, but preferred standard RSVP Keyboard. Inquiry Preview is a novel multimodal interface for the RSVP Keyboard BCI, incorporating switch input as an additional control signal. Typing performance and user experience and preference varied widely across participants, reinforcing the need for flexible, customizable BCI systems that can adapt to individual users. gov Identifier: NCT04468919.

A Comparison of Glucose and Additional Signals for Three Different Exercise Types in Adolescents with Type 1 Diabetes Using a Hybrid Closed-Loop System.

Objective: To compare glycemic outcomes during and following moderate-intensity exercise (MIE), high-intensity interval exercise (HIE), and resistance exercise (RE) in adolescents with type 1 diabetes (T1D) using a hybrid closed-loop (HCL) insulin pump while measuring additional physiological signals associated with activity. Methods: Twenty-eight adolescents (average age 16.3 ± 2.1 years, 50% females, average duration of T1D 9.4 ± 4 years) using HCL (Medtronic MiniMed 670G) undertook 40 min of MIE, HIE, and RE. A temporary glucose target (8.3 mmol/L, 150 mg/dL) was set for 2 h prior and during exercise. Heart rate, accelerometer, venous glucose, lactate, ketones, and counter-regulatory hormones were measured for 280 min postexercise commencement. The primary outcome was glucose percentage time in range (TIR): 3.9-10 mmol/L (70-180 mg/dL) for 14 h from exercise onset. Results: Median (interquartile range) TIR for HIE was 88 (78, 96)%, MIE 79 (63, 88)%, and RE 86 (72, 95)% for 14 h from exercise onset. For MIE compared with HIE, TIR was lower (P = 0.012) and time above range (TAR) was greater (18 [2.4, 28] vs. 6.9 [0.0, 14]%, P = 0.041). Hypoglycemia occurred in 13 (46%), 11 (39%), and 14 (50%) of participants for HIE, MIE, and RE, respectively, the majority following the meal after exercise. There were higher levels of lactate (P = 0.001), growth hormone (P = 0.001), noradrenaline (P = 0.001), and heart rate (P = 0.01) during HIE and RE compared with MIE. Conclusions: HCL use in adolescents with T1D results in excellent TIR during different forms of exercise when a temporary target is set 2 h before. Extending the temporary target after exercise may also be needed to help minimize postexercise hypoglycemia.

PPARα agonist ameliorates cholestatic liver injury by regulating hepatic macrophage homeostasis.

Inflammatory response plays an essential role in the pathogenesis of cholestatic liver injury. PPARα agonists have been shown to regulate bile acid homeostasis and hepatic inflammation. However, the immunoregulatory mechanisms through which PPARα agonists ameliorate cholestatic liver injury remain unclear. In this study, surgical bile duct ligation was performed to establish a mouse model of cholestasis. Our study revealed that PPARα agonist alleviated cholestatic liver injury in mice by suppressing inflammatory response, reducing neutrophil infiltration, and promoting M2-like macrophage polarization. CyTOF analysis showed that PPARα agonist increased the proportion of anti-inflammatory F4/80hiCD44+MHCII- M2-like macrophages while decreasing the proportion of pro-inflammatory CD64+CX3CR1+CCR2hiVISTAhiCD172a+CD44hi M1-like MoMFs. Additionally, scRNA-seq indicated that PPARα agonist regulated the developmental trajectory and homeostasis of hepatic macrophages. Mechanistically, PPARα agonist may influence the expression of immune regulators in heterogeneous macrophages to exert protective effects against cholestasis. In addition, the CCL and MIF signaling pathways may participate in the communication among hepatic immune cells, including macrophages, neutrophils, natural killer cells, and dendritic cells, in response to the PPARα agonist. In conclusions, PPARα agonist alleviated cholestatic liver injury by attenuating the inflammatory response and restoring hepatic macrophage homeostasis. This study might enhance the understanding of the immunoregulatory mechanisms of PPARα agonists, providing promising therapeutic targets for cholestatic liver diseases.

Increasing the Level of Knock-in of a Construct Encoding the HIV-1 Fusion Inhibitor, MT-C34 Peptide, into the <i>CXCR4</i> Locus in the CEM/R5 T Cell Line

The low efficiency of knock-in, especially in primary human cells, limits the use of genome editing technology for therapeutic purposes, which makes it important to develop approaches for increasing knock-in levels. In this work, using a knock-in model of the peptide fusion inhibitor of HIV MT-C34 into the human CXCR4 locus in the CEM/R5 T cell line, we analyzed the effectiveness of several approaches to increasing knock-in levels. First, donor DNA modification aimed at improving the efficiency of plasmid transport into the nucleus was evaluated, namely the introduction into the donor plasmid of the SV40 DNA transport sequence (DTS) or the binding sites for the transcription factor NF-κB, whose effects on knock-in levels have not been described. In the MT-C34 knock-in model into the CXCR4 locus, this modification was ineffective. The second approach, modifying the Cas9 nuclease by introducing two additional nuclear localization signals (NLS), increased the knock-in level by 30%. Finally, blocking DNA repair via the nonhomologous end joining pathway using DNA-dependent protein kinase inhibitors caused a 1.8-fold increase in knock-in. The combination of the last two approaches caused an additive effect. Thus, increasing the number of NLSs in the Cas9 protein and inhibiting DNA repair via the nonhomologous end joining pathway significantly increased the level of knock-in of the HIV-1 peptide fusion inhibitor into the clinically relevant locus CXCR4, which can be used to develop effective gene therapy approaches for the treatment of HIV infection.

Allium chromosome evolution and DNA sequence localization.

Molecular cytogenetics, utilizing DNA probes, serves as a critical tool for mapping genes to the physical structures of chromosomes. In this study, we examined three Allium species: A. cepa L., A. sativum L., and A. fistulosum L., using in situ hybridization to localize 45S rDNA and 5S rDNA genes. We observed variation in both the chromosomal localization and signal intensity of the 45S and 5S rDNA probes across the species. Notably, in A. sativum, additional 5S rDNA signals were detected on chromosome 8, in a heterozygous condition. Additionally, we aimed to explore the feasibility of localizing genes associated with pigment biosynthesis in A. cepa, specifically the PAL and FLS genes. For this, we employed TSA-FISH on both meiotic and mitotic chromosomes. Preliminary results suggested that the PAL gene was localized to meiotic metaphase chromosomes, while the single-copy FLS gene was detected on mitotic chromosomes. The TSA-FISH technique proved neither routine nor robust for consistent localization of these specific probes in plant chromosomes. The findings based on rDNA analysis also offer insights into potential evolutionary implications among the different Allium species studied.

Targeting signals required for protein sorting to sub-chloroplast compartments.

Chloroplasts, distinctive subcellular organelles found exclusively in plant species, contain three membranes: the outer, inner, and thylakoid membranes. They also have three soluble compartments: the intermembrane space, stroma, and thylakoid lumen. Accordingly, delicate sorting mechanisms are required to ensure proper protein targeting to these sub-chloroplast compartments. Except for most outer membrane proteins, chloroplast interior proteins possess N-terminal cleavable transit peptides as primary import signals. After the cleavage of transit peptides, which occurs during or after import into chloroplasts, the inner and thylakoid membrane proteins, as well as stromal and thylakoid luminal proteins, are further sorted based on additional targeting signals. In this review, we aim to recapitulate the mechanisms by which proteins are targeted to chloroplasts and subsequently sorted into sub-chloroplast compartments, with a focus on the design principles of sorting signals present in chloroplast proteins.

Characteristics and cellular and molecular biomechanical influencing factors of resilience: A cross-sectional study of nurses experiencing workplace violence in Jiangsu China

Objective: To investigate the resilience status of nurses after workplace violence and its influencing factors from the perspective of cellular and molecular biomechanics. Methods: From April to July 2024, a cross-sectional questionnaire study was conducted utilizing the General Information Questionnaire, the Medical Staff Resilience Scale (MSRS), the Workplace Violence Scale (WVS), the General Self-Efficacy Scale (GSE), and the Perceived Social Support Scale (PSSS) on a sample of 375 nurses who had been victims of workplace violence at six tertiary-level A general hospitals and three secondary-level hospitals in Jiangsu Province, China. The data were subsequently analyzed. Results: The nurses’ resilience score was (72.37 ± 10.19) with a mean score of (4.02 ± 0.57). Multiple regression analysis showed that age, work experience, title, monthly income, self-efficacy, and social support independently influenced their resilience. (P < 0.05). Pearson’s correlation analysis revealed a positive correlation between carers’ levels of resilience with generic self-efficacy scale and social support (P < 0.01), and a negative correlation of resilience with workplace violence (P < 0.01). Conclusion: The resilience scores of nurses who had suffered workplace violence were found to be at a moderate level. Stressors from workplace violence might trigger complex intracellular signaling pathways and molecular changes in nerve cells and endocrine cells of nurses. Hormonal imbalances could further affect neurotransmitter systems and molecular cascades related to mood regulation and stress adaptation, thereby influencing the nurses’ resilience. Higher self-efficacy could potentially enhance the activation of positive molecular pathways and the expression of certain genes related to stress resistance. Social support might buffer the negative impacts of workplace violence on cellular and molecular mechanisms by providing additional resources and positive molecular signals. It is recommended that managers consider the effects of age, years of work experience, job title, and monthly income when developing strategies to enhance resilience. It is also the responsibility of managers to facilitate the mobilization of resources, both internal and external, related to self-efficacy and social support. Furthermore, they should construct targeted training courses based on an analysis of the mechanisms involved in the occurrence of workplace violence, to improve the resilience of nurses.

EOMES establishes mesoderm and endoderm differentiation potential through SWI/SNF-mediated global enhancer remodeling.

Mammalian pluripotent cells first segregate into neuroectoderm (NE), or mesoderm and endoderm (ME), characterized by lineage-specific transcriptional programs and chromatin states. To date, the relationship between transcription factor activities and dynamic chromatin changes that guide cell specification remains ill-defined. In this study, we employ mouse embryonic stem cell differentiation toward ME lineages to reveal crucial roles of the Tbx factor Eomes to globally establish ME enhancer accessibility as the prerequisite for ME lineage competence and ME-specific gene expression. EOMES cooperates with the SWItch/sucrose non-fermentable (SWI/SNF) complex to drive chromatin rewiring that is essential to overcome default NE differentiation, which is favored by asymmetries in chromatin accessibility at pluripotent state. Following global ME enhancer remodeling, ME-specific gene transcription is controlled by additional signals such as Wnt and transforming growth factor β (TGF-β)/NODAL, as a second layer of gene expression regulation, which can be mechanistically separated from initial chromatin remodeling activities.

Single photon detection based on photon driven mode for improving the depth reconstruction quality

Abstract In the single photon detection, the precise time of flight (TOF) between the target and the detector is very important to retrieve the depth information with high quality. It is hard to obtain in the practical experiment due to the existence of the system jitter, the background noise and the dead time, respectively. In this letter, a single photon detection method based on photon driven mode is proposed to improve the depth reconstruction quality. An additional reference signal is added to suppress the delay of the system. The photon driven model is built to explore effects of the dead time on the incident photon sequence. Finally, the incident photon sequence retrieved by maximum likelihood estimate (MLE) is filtered by a butterworth filter to improve the depth information.

Lightning and Radar Measures of Mixed-Phase Updraft Variability in Tracked Storms during the TRACER Field Campaign in Houston, Texas

Abstract Properties of 7488 thunderstorms are summarized for June–September 2022 during the Tracking Aerosol Convection Interactions Experiment (TRACER) field campaign Houston, Texas, using polarimetric weather radar and VHF 3D Lightning Mapping Array data. Automated tracking of storms linked each instrument’s measurements to a data-defined, time-evolving storm footprint. Within each storm, the depth and magnitude of episodic columns of radar differential reflectivity and specific differential phase quantified the prevalence of updrafts that activated mixed-phase precipitation pathways. Lightning measurements further distinguished the degree of rimed precipitation formation: the fraction of tracks with lightning varied from day to day and cells with lightning had stronger polarimetric columns. Track-level correlation of the lightning flash rate with radar polarimetric measures had substantial spread, showing that lightning provides an additional signal of mixed-phase precipitation processes that can complement future studies of thermodynamic and aerosol controls on cloud microphysics in the Houston region.

Mogamulizumab-Associated Autoimmune Diseases: Insights From FAERS Database Analysis.

Mogamulizumab is a monoclonal antibody targeting the C-C chemokine receptor 4, used to treat T-cell malignancies such as cutaneous T-cell lymphoma, adult T-cell leukemia/lymphoma, and peripheral T-cell lymphoma. However, real-world studies on mogamulizumab-associated adverse events (AEs) are limited. Disproportionality analyses were performed to assess the safety profile of mogamulizumab based on data from the US Food and Drug Administration Adverse Event Reporting System (FAERS) database for the period spanning from October 2018 to December 2023. The research investigated demographic characteristics, the onset timing of AEs, and the safety implications associated with mogamulizumab use. A total of 1182 significant preferred terms were identified among the 3661 mogamulizumab-associated AE reports collected from the FAERS database. The frequently reported AEs including rash, infusion-related reaction, and pyrexia were in line with drug instruction. Notably, several unexpectedly significant AEs were also found, including pemphigoid (ROR = 5.69 [95% CI 1.83-17.66]), unstable angina (ROR = 20.56 [95% CI 8.54-49.5]), bulbar palsy (ROR = 238.36 [95% CI 75.22-755.31]), myositis (ROR = 12.65 [95% CI 5.67-28.19]), and various autoimmune diseases such as autoimmune hepatitis (ROR = 21.33 [95% CI 11.08-41.07]), myocarditis (ROR = 15.29 [95% CI 8.67-26.97]), glomerulonephritis (ROR = 22.49 [95% CI 7.24-69.9]), nephrotic syndrome (ROR = 7.63 [95% CI 2.46-23.67]), myasthenia gravis (ROR = 8.54 [95% CI 3.2-22.77]), and autoimmune thyroiditis (ROR = 11.81 [95% CI 3.8-36.68]). This study replicated previously identified AEs associated with mogamulizumab and uncovered additional signals of AEs, particularly emphasizing the risks associated with autoimmune diseases. It is essential to exercise vigilance in monitoring the occurrence of these AEs during the use of mogamulizumab in clinical practice.

Enhancement of cyclic spectral coherence map by statistical testing approach—application to bearing faults diagnosis in electric motors

Abstract Efficiency of fault detection in rolling element bearings is heavily influenced by the quality of data. In controlled environments, such as test rigs designed for bearing diagnostics, data quality is relatively good. Similarly, diagnosing bearings that support shafts in industrial machinery is relatively straightforward. However, diagnosing bearings in electric motors presents greater complexity due to the influence of additional cyclic components on vibration signals. These extra components, originating from mechanical or electrical sources, complicate frequency-based analysis. This paper proposes a novel approach for diagnosing bearings in electric motors, utilizing statistical analysis within the bi-frequency domain through a cyclostationary framework. The method involves applying a statistical testing procedure to individual pixels on the cyclic spectral coherence (CSC) map. The statistical significance of these pixels is assessed based on quantiles of CSC maps obtained from a dataset representing a healthy bearing. This process results in an enhanced or cleaned CSC map, facilitating the identification of fault-related components. Consequently, this approach enables the detection of defects in electric motor bearings, even when additional signal components unrelated to the defect, but characteristic of a healthy bearing, are present.

More than stridulation: signal interaction and constraint in the complex vibroacoustic courtship of a cricket

Field crickets (Gryllidae) produce sounds by tegminal stridulation, well-studied for its role in female attraction and choice. However, understanding female preferences for their courtship song remains elusive, despite considering additional chemical, visual, and thermal signals. Beyond stridulation, crickets also display vibrational courtship behaviours that remain largely unexplored. Using Acheta domesticus as a model, we conduct the first comprehensive analysis of the entirety of vibroacoustic courtship signals in crickets, including their interaction. Employing audio recording, laser vibrometry, and videorecording, we unveil a complex signal involving simultaneous wing stridulation, body tremulation, and leg drumming against the substrate in a prolonged display, unique among insects. We identify robust correlations, coupling, and coordination between these signal components. We show the tightest coupling between the two types of stridulation pulses, and between tremulation and drumming signals, while drumming-stridulation coupling is less consistent, revealing a constraint on drumming performance. This constraint in the expression of one signal component, without a trade-off, represents a specific case within complex dynamic signalling. In addition, we find no correlation between drumming rate and its accuracy relative to stridulation, challenging common expectations. Our findings indicate that the information conveyed by the complex courtship display in A. domesticus is not simply proportional to that in the song, shedding light on previous ambiguities surrounding its function. Spectral-intensity analysis indicates the closest perceptual connection between stridulation and drumming signals, likely commonly influencing female choice, while proposing another function for tremulation. Further research should delve deeper into the function of this intricate signal.

A memristor‐sourced key for encrypted power and data synchronous transmission in switched mode power supplies

AbstractIn power and data synchronous transmission (PDST), it is crucial to modulate additional signals onto the voltage bus of the converters. This reduces the cost of monitoring and control of the power grid. There is thus the need to ensure that the data transmitted is well secured. This study, designs an electronic system that uses the common coupling point for PDST. Then Chua's memristive circuit is introduced as an entropy source for generating a random bit sequence for encrypted PDST in a switched mode power supply. The encrypted data signal is modulated using phase shift key modulation. Thereafter, the carrier signal is pulse width modulation modulated to control the switching operation of the power switch. The use of Chua's memristive circuit for generating a random number is not new; however, there is little study directly adopting its use in PDST. This allows for resource sharing since multiple signal sources can share the same bus without interference or security risk. The generated random bits are tested for randomness using National Institute of Standards and Technology test suite and autocorrelation test. MATLAB/Simulink and experimental results both confirm the effectiveness of the proposed data encryption method.

Sodium-dependent glucose co-transport proteins (SGLTs) are not involved in human glucose taste detection

The sweet taste of saccharides, such as sucrose and glucose, and other sweeteners is known to result from activation of the TAS1R2/R3 receptor expressed in taste receptor cells (TRCs) of the taste bud. Recent reports have suggested the existence of an additional sweet taste signaling pathway for metabolizable saccharides that is dependent on the activity of glucose transporters, especially SGLT1, also expressed in TRCs. We have investigated the potential contribution of SGLT1 to glucose taste signaling in humans. Concentration-response analysis of glucose mediated changes in membrane potential measured in Chinese hamster ovary (CHO) cells transiently expressing the human SGLT1 (hSGLT1) yielded an EC50 value of 452 μM. The SGLT inhibitor phlorizin inhibited the membrane potential response to 10 mM glucose with an IC50 of 3.5 μM. In contrast, EC50 values of 127 and 132 mM were obtained from concentration-response analysis of glucose taste in vehicles of water or 20 mM NaCl, respectively, by rapid throughput taste discrimination with human subjects. Lactisole, an antagonist of TAS1R2/R3, at a concentration of 1 mM completely inhibited taste responses to glucose concentrations of 250 mM and below. Phlorizin (0.2 mM) and the high potency SGLT1-selective inhibitor mizagliflozin (10 μM) failed to inhibit glucose taste detection measured at peri-threshold concentrations in the rapid throughput taste discrimination assay. A Yes/No experiment using the taste discrimination assay revealed that 0.2 mM phlorizin was discriminable from water for some subjects. Taken together the results indicate that agonist activation of TAS1R2/R3 is sufficient to account for all glucose taste without contribution by an alternative SGLT-mediated signaling pathway. Furthermore, the taste of phlorizin could be a confounding variable for studies evaluating a role for SGLTs in taste.

Dynamics of convolutional recurrent neural networks near their critical point

We examine the dynamical properties of a single-layer convolutional recurrent network with a smooth sigmoidal activation function, for small values of the inputs and when the convolution kernel is unitary, so all eigenvalues lie exactly at the unit circle. Such networks have a variety of hallmark properties: the outputs depend on the inputs via compressive nonlinearities such as cubic roots and both the and the depend sensitively on the inputs as power laws, both diverging as the input →0. The basic dynamical mechanism is that inputs to the network generate ongoing activity, which in turn controls how additional inputs or signals propagate spatially or attenuate in time. We present analytical solutions for the steady states when the network is forced with a single oscillation and when a background value creates a steady state of “ongoing activity” and derive the relationships shaping the value of the temporal decay and spatial propagation length as a function of this background value. Published by the American Physical Society 2024

CAUSALdb2: an updated database for causal variants of complex traits.

Unraveling the causal variants from genome wide association studies (GWASs) is pivotal for understanding genetic underpinnings of complex traits and diseases. Despite continuous efforts, tools to refine and prioritize GWAS signals need enhancement to address the direct causal implications of genetic variations. To overcome challenges related to statistical fine-mapping in identifying causal variants, CAUSALdb has been updated with novel features and comprehensive datasets, morphing into CAUSALdb2. This expanded repository integrates 15057 updated GWAS summary statistics across 10839 unique traits and implements both LD-based and LD-free fine-mapping approaches, including innovative applications of approximate Bayes Factor and SuSiE. Additionally, by incorporating larger LD reference panels such as TOPMED and UK Biobank, and integrating functional annotations via PolyFun, CAUSALdb2 enhances the accuracy and context of fine-mapping results. The database now supports interrogation of additional causal signals and offers sophisticated visualizations to aid researchers in deciphering complex genetic architectures. By facilitating a deeper and more precise characterisation of causal variants, CAUSALdb2 serves as a crucial tool for advancing the genetic analysis of complex diseases. Available freely, CAUSALdb2 continues to set benchmarks in the post-GWAS era, fostering the development of targeted diagnostics and therapeutics derived from responsible genetic research. Explore these advancements at http://mulinlab.org/causaldb.

Abstract PR017: Anti-metastatic immunotherapy discovery using ex vivo lung tissue cultures and high-throughput single-cell chemical transcriptomics

Abstract Introduction: Tumors systemically remodel the immune system during metastasis. Developing anti-metastatic immunotherapies that target these tumor-immune interactions to make the metastatic niche hostile colonization would represent a paradigm shift in cancer treatment. However, traditional high-throughput screening (HTS) platforms that use simple read-outs and contrived in vitro systems are ill-suited to identify such therapies. In contrast, HTS platforms that accurately model the metastatic niche and use high-dimensional read-outs such as multiplexed single-cell RNA-sequencing (scRNA-seq) can accurately profile nuanced perturbation responses in individual immune cell types and therefore have the potential to discover anti-metastatic immunotherapies. In this study, we describe the development of an ex vivo lung tissue culture HTS platform that preserves immune gene expression signatures observed in the in vivo metastatic niche and is amenable to single-cell chemical transcriptomic analysis using MULTI- seq (McGinnis et al., Nature Methods, 2019). Inspired by our recent description of myeloid cell TLR-NFκB inflammation during breast cancer lung metastasis (McGinnis et al., Cancer Cell, 2024), we then performed the first anti-metastatic immunotherapy drug screen and identified TLR-NFκB inhibitors that effectively operate on all desired myeloid cell types in the lung metastatic niche. Methods and Results: Building upon existing techniques for culturing precision-cut lung slices (Wu et al., Journal of Experimental Medicine, 2024) and patient- derived tumor fragments (Voabil et al., Nature Medicine, 2021), we established and optimized an ex vivo lung tissue culture system that enables 48-hour culturing of lung slices isolated from 4T1 tumor-bearing mice. scRNA-seq profiling revealed successful capture of all relevant immune cell types and retention of metastasis-associated gene expression programs (e.g., myeloid TLR-NFκB inflammation and neutrophil degranulation) following culture. We then scaled our platform to address how myeloid cells in the lung metastatic niche respond to perturbations targeting every component of the canonical TLR-NFκB signaling cascade. Cell-type-specific perturbation modeling and quantification of TLR-NFκB signaling inhibition identified compounds that optimally perturb all intended myeloid cell types (e.g., tissue-resident macrophages, neutrophils, and monocytes) and represent prioritized candidates for future in vivo validation studies to assess impact on metastatic disease progression. Conclusions: In this study, we describe the development and application of a novel HTS platform that couples ex vivo lung tissue cultures with single-cell chemical transcriptomics to identify anti-metastatic immunotherapy candidates. We demonstrate how our platform can identify TLR-NFκB signaling inhibitors that optimally operate in the lung metastatic niche, paving the way for future interrogation of additional metastasis-associated immune cell signaling pathways in different disease and tissue backgrounds. Citation Format: Chris McGinnis, Winnie Yao, Ansuman Satpathy. Anti-metastatic immunotherapy discovery using ex vivo lung tissue cultures and high-throughput single-cell chemical transcriptomics [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr PR017.

Intra- and inter-patient diversity in hepatocellular carcinoma based on phosphorylation profiles—A pilot study in a single institution

BackgroundRecent studies have addressed the efficacy of targeted drugs against hepatocellular carcinoma. However, most tumors escape a single kinase inhibition; co-inhibition of additional signaling pathways re-sensitizes resistant cancer cells to targeted drugs, thus reinforcing the importance of combination therapy for drug-resistant tumors. This study aimed to clarify the phosphorylation profiles of representative cancer-related tyrosine kinases in hepatocellular carcinoma to focus on potential therapeutic targets and to investigate the possibility of expanding combination therapy options using targeted drugs. Materials and MethodsPatients’ whole blood, hepatocellular carcinoma tissue, and adjacent hepatic tissues were obtained during surgeries from 10 patients. All patients showed negative results for hepatitis B and hepatitis C RNA and none had a history of heavy drinking. The activation of receptor tyrosine kinases (RTKs) was analyzed by using a human RTK phosphorylation antibody array. ResultsAmong 62 different phospho-RTKs, 26 were activated in tumor tissues, of which ACK1, Dtk, Fyn, and Lyn were positive in 9 out of 10 cases. The median concordance rates of activated tumor and serum RTKs in each patient was 50 %. There was an inter- and intra-patient diversity of phosphorylation profiles in the serum, tumor of resected specimens, and non-tumor tissue of resected specimens in the same patients. ConclusionThere was an intra- and inter- patient diversity in the activation of important and representative cancer-related RTKs. Expanding on this approach will allow us to learn how to predict the best combination of targets for each patient and to prioritize those combinations for clinical testing.

A PSMA-targeted Tri-specific Killer Engager enhances NK cell cytotoxicity against prostate cancer.

Natural killer (NK) cell tumor infiltration is associated with good prognosis in patients with metastatic castration-resistant prostate cancer (mCRPC). NK cells recognize and kill targets by a process called natural cytotoxicity. We hypothesized that promoting an antigen-specific synapse with co-activation may enhance NK cell function in mCRPC. We describe a Tri-specific Killer Engager (TriKE) construct that engages with the activating receptor CD16 on NK cells, prostate-specific membrane antigen (PSMA) on mCRPC cells, and has an interleukin (IL)-15 moiety that is essential for NK cell survival, proliferation, and priming. We show that the PSMA TriKE specifically binds to PSMA-expressing cells and significantly enhances expansion, degranulation and cytokine production of NK cells derived from healthy donors or prostate cancer patients. Bystander killing of PSMA-negative was also achieved with PSMA TriKE treatment when co-cultured with PSMA-positive cells, suggesting potential PSMA TriKE benefit in controlling tumor antigen escape. When tested under physiologic conditions recapitulating the mCRPC tumor microenvironment (TME), NK cells treated with PSMA TriKE and prolonged exposure to hypoxia or MDSCs maintained their potent function while IL-15 treated NK cells showed greatly impaired cytotoxicity. Finally, in vivo testing of PSMA TriKE showed improved tumor control and survival of mice as compared to IL-15 and untreated control groups. In conclusion, PSMA TriKE demonstrates potential as a new therapy for advanced prostate cancer by providing additional signals to NK cells to maximize their anti-tumor potential in prostate cancer, especially in the setting of a hostile TME.