Weiqiang Chen

Vaccines have revolutionized public health, yet their development remains hampered by the poor predictive power of animal models, leading to high clinical failure rates and variable efficacy across populations. To bridge this translational gap, we developed a human lymph node-on-a-chip model that biomimetically reproduces key physiological features of human lymph node, including compartmentalized immune cell zones and functional stromal networks. This immunocompetent model recapitulates the complete cascade of vaccine-induced adaptive immune processes in human lymph nodes, from antigen presentation, immune cell differentiation to germinal center formation and antibody secretion, providing a human-relevant platform for preclinical vaccine testing. Using donor-matched immune cells from influenza vaccine trials, we established a personalized clinical-trial-on-chip platform that accurately mirrors individual vaccine responses. Benchmarking on-chip immunogenicity readouts against clinical serological and transcriptomic data confirmed the platform’s predictive power for assessing vaccine efficacy across diverse populations. Our study uncovered a key mechanism of vaccine failure in vulnerable populations: age- and comorbidities-related factors impair T follicular helper cell differentiation and disrupt critical T-B cell interactions. Single-cell transcriptomic profiling revealed critical immune signaling networks involving MyD88 in DCs, IL-2/STAT5 balance in T cells, and TACI/BCMA activation in B cells that collectively govern the efficiency of the adaptive immune cascade. These mechanistic insights enabled us to validate clinically actionable strategies, including dose escalation and IL-2 cytokine adjuvant as effective countermeasures to enhance immunogenicity efficiency in immunocompromised individuals. These results demonstrate the potential of this human lymph node-on-a-chip as a transformative precision vaccinology tool for personalized vaccine immunogenicity assessment and optimization.

Survival demands instant prioritization of escape over maintenance. To decode the dynamic logic embedded in the static Drosophila connectome (FlyWire v783), we simulated the conflict between predator-evasion, feeding, and grooming to uncover the logic of this switch. We show that escape is a holistic state defined by distributed robustness: it is reliably triggered by only a fraction of visual inputs (LC4/LPLC2) and induces a system-wide pause on behaviors like feeding and grooming. Within this global suppression, we identify a specialized, fail-safe architecture for arresting feeding. A redundant ensemble of neurons (DNge031/CB0565) targets the premotor center Roundup, effectively disfacilitating motor output. Crucially, this inhibition is modular, anatomically distinct from grooming suppression, and relies on additive logic to ensure suppression even if individual components fail. Our results demonstrate how connectome topology implements robust, survival-critical control through distributed neural architecture.

Given selective breeding has resulted in pronounced differences in muscle development and production traits among broilers, layers, and silkies, this study systematically compared breed-specific variations in growth performance, serum biochemical markers, and duodenal glucose transporter (GLUT) expression under basal and insulin-stimulated conditions. Herein, broilers exhibited the fastest growth and greatest muscle accretion, accompanied by higher basal insulin and uric acid, but lower blood glucose, urea, albumin, triglycerides, and low-density lipoprotein cholesterol (LDL-c) compared to layers and silkies. Layers had higher basal blood glucose and distinctive duodenal upregulation of GLUT3, GLUT4, GLUT5, GLUT8, and GLUT9, fitting their high energy demands for egg production. Silkies displayed unique serum protein profiles (increased total protein and globulin) and an intermediate metabolic phenotype. Notably, broilers showed a marked, sustained hypoglycemic response and elevated insulin after exogenous insulin administration, which coincided with rapid, transient upregulation of duodenal GLUT1, GLUT3, GLUT8, GLUT9, and GLUT12. In contrast, layers and silkies showed milder and breed-specific changes in serum biomarkers and GLUT expression, with silkies exhibiting delayed but persistent GLUT9 induction and partial suppression of certain GLUTs after insulin. These findings demonstrate that breed-specific patterns of serum biochemistry and intestinal GLUT regulation are closely aligned with physiological and production adaptations in each breed. In particular, rapid growth in broilers is likely driven by enhanced insulin sensitivity and dynamic GLUT response for efficient glucose uptake, while layers and silkies employ alternative strategies for nutrient absorption and metabolism.

CAR T cell cytotoxicity requires generating immense mechanical force, but the energetic costs of this process remain poorly defined. While metabolic reprogramming fuels effector function, its mechanistic connection to mechanotransduction remains unclear. By directly measuring the synaptic force and mechanical energy of single CAR T cells and linking them to their metabolic state, we proved that the mechano-energetic efficiency is a fundamental determinant of cytotoxic potency. We discovered that the mechanosensitive ion channel Piezo1 couples cytoskeletal dynamics to metabolic rewiring via Ca2+-Wnt-Rac1 signaling. Disrupting Piezo1 cripples glycolytic and mitochondrial ATP production, causing energetic stress and impaired cytotoxicity. Notably, Piezo1 activity follows a Goldilocks principle: intermediate level maximizes activation and cytotoxicity, whereas either hypoactive or hyperactive Piezo1 states impair mechano-metabolic fitness and drive dysfunction in patient and exhausted CAR T cells. Our work establishes mechano-metabolic coupling as a core regulator of CAR T cell fitness and pinpoints Piezo1 tuning as a new strategy to enhance cancer immunotherapy.

Human dermcidin (DCD) is synthesized as a 110-amino acid precursor (pre-dermcidin, pre-DCD) containing a 19-residue leader signal sequence, which is removed to produce a leader-less pro-domain-containing peptide termed as pro-dermcidin, pro-DCD. Pro-DCD can be secreted by human eccrine sweat glands and then cleaved into antimicrobial peptides, such as dermcidin (DCD). Emerging evidence suggests that pro-DCD has broader physiological roles beyond antimicrobial defense, potentially serving as a therapeutic agent for inflammatory diseases like sepsis. In this review, we summarize recent evidence supporting pro-DCD as a regulator of innate immunity in sepsis.

The MAST family of serine/threonine kinases has been implicated in a spectrum of human neurodevelopmental disorders. However, little is known about their biological function or regulation. Seeking to fill these gaps in our knowledge, we have identified upstream and downstream partners of MAST1. 14-3-3η, a neuronal 14-3-3 paralog, specifically interacts with MAST1 at two regulatory serines, S90 and S161. PAK, a neuronal regulator of the actin cytoskeleton, phosphorylates MAST1 to regulate its interaction with 14-3-3η. Exploiting mouse models of human Mega Corpus Callosum Syndrome (MCC) and whole brain phosphoproteomics, we identify the microtubule-associated protein Tau as a substrate of MAST1. We show that pathogenic MAST1 mutations perturb protein function either through misfolding or attenuation of kinase activity. Our data is consistent with a model in which the MAST kinases couple PAK, a neuronal regulator of the actin cytoskeleton, to microtubule remodeling during the differentiation and specification of cortical neurons.

Chimeric antigen receptor (CAR) T cell immunotherapy is promising for treatment of blood cancers; however, clinical benefits remain unpredictable, necessitating development of optimal CAR T cell products. Unfortunately, current preclinical evaluation platforms are inadequate owing to their limited physiological relevance to humans. Here we engineer an organotypic immunocompetent chip that recapitulates microarchitectural and pathophysiological characteristics of human leukaemia bone marrow stromal and immune niches for CAR T cell therapy modelling. This leukaemia chip empowers real-time spatiotemporal monitoring of CAR T cell functionality, including T cell extravasation, recognition of leukaemia, immune activation, cytotoxicity and killing. We use our chip to model clinically observed heterogeneous responses such as remission, resistance and relapse under CAR T cell therapy and map factors that drive therapeutic success or failure. Finally, we demarcate functional performance of CAR T cells produced from different healthy donors and patients with cancer, with various CAR designs and protocols, systematically and multidimensionally. Together, our chip introduces an enabling '(pre-)clinical-trial-on-chip' tool for CAR T cell development, which may translate to personalized therapies and improved clinical decision-making.

Andrographolide is a prominent labdane diterpenoid extracted from Andrographis paniculata with exceptional anti-inflammatory properties. Commercial production of andrographolide relies exclusively on extraction from plant resources. Although the scaffold of andrographolide ent-copalol has previously been biosynthesized, further oxidative modifications remain elusive. In this study, by taking an integrated analysis of transcriptomes and metabolomes, we were able to identify four cytochrome P450 enzymes constituting the minimal set of andrographolide biosynthetic genes. Specifically, ApCYP71D587 catalyzes the conversion of ent-copalol to 19-hydroxy-ent-copalol. Subsequently, ApCYP71BE50 mediates the formation of the lactone ring, ultimately yielding andrograpanin. Then ApCYP706U5 accomplishes the third step by mediating the C-3 hydroxylation reaction, thereby allowing the formation of 14-deoxyandrographolide. Ultimately, ApCYP72F1 completes the biosynthetic generation of andrographolide with C-14 hydroxylation of the lactone and rearrangement of the olefin bond. In addition, co-expression of the minimal gene set in N. benthamiana engineered to produce ent-copalol feasibly produces andrographolide, thus establishing an innovative metabolic engineering strategy to produce this medicine of historical importance, circumventing the need for plant extraction.

Abstract Purpose: Radiation- and contrast-free magnetic resonance imaging (MRI) may facilitate investigations of early-life and longitudinal airway tree structure, but manual segmentations can be time consuming. Leveraging recent advances in self-supervised learning, without the need of manual labeled MRI, we propose an automated framework to segment the central airway tree from MRI of adolescents. Methods and Materials: We acquired pulmonary MRI with voxel size 1.3x1.4x1.3 mm³ including Zero Echo Time, Ultrashort Echo Time (UTE), UTE with advanced motion correction, and LAVA sequence (voxel size 0.9x2.0x0.9 mm³) for 44 adolescents and young adults (34 for training and 10 for evaluation). We employed unsupervised domain adaptation to transfer segmentation knowledge learned from the source domain (n=280 CTs and masks, voxel size 0.7x0.7x0.5 mm³ from the Airway Tree Modeling Challenge 2022, using airway mask from trachea to segmental bronchi) to the target domain (n=136 lung MRIs only, from 34 training subjects, in a self-supervised manner). Using our Masked Autoencoding and Pseudo-labeling Segmentation (MAPSeg) method, the model first learned and performed whole-lung MRI segmentation. The MRIs were then cropped to the lung mask bounding box to target airway segmentation. Model estimated segmentations were compared to manual segmentations on UTE MRI with advanced motion correction in a test dataset (n=10) using dice similarity coefficient (DSC), average surface distance (ASD), precision, sensitivity, specificity, and lumen diameters averaged across the middle 2/3 of each airway segment. Results: The 44 participants had a mean±SD age of 19.8±1.2 years old, and 52% were female. Manual and model segmentations of airway tree yielded DSC 0.83±0.02, ASD 0.75±0.04 mm, precision 0.89±0.03, sensitivity 0.78±0.03, and specificity 0.99±0.00. Both manual segmentations and model captured all trachea, mainstem and lobar airways. The manual vs. model estimated mean of airway lumen diameters for trachea to lobar airways was 9.0±1.5mm vs. 8.6±1.3mm, with a Pearson correlation r=0.96 (P<0.001), and by anatomical level: trachea: 13.4±2.7mm vs. 12.7±2.8mm (r=0.99, P<0.001); mainstems: 10.4±1.7mm vs. 9.7±1.6mm (r=0.95, P=0.004); and lobar: 7.3±1.4mm vs. 7.1±0.9mm (r=0.84, P=0.03). Manual and model segmentation identified 5.6±3.1 vs. 2.0±2.1 segmental bronchi in 6 participants, with lumen diameter 4.4±0.7mm vs. 5.3±0.6mm (r=0.89, P=0.01). Conclusion: The preliminary model trained via MAPSeg demonstrated excellent detection and lumen diameter accuracy for automated segmentation of trachea to lobar airways on MRI images, with lower airway detection but preserved lumen diameter accuracy at the segmental level. In future studies, we will optimize the model for segmental airways.

Chimeric antigen receptor (CAR) T-cell therapy has demonstrated promising outcomes in treating leukemia, yet it confronts significant hurdles due to severe side effects like cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Furthermore, existing preclinical evaluation platforms often fall short in accurately replicating the human tumor microenvironment, leading to extended development durations and elevated costs. To overcome these challenges, we have developed an innovative, integrated microfluidic chip equipped with microarchitectural features that precisely mimic tumor niches, thus enhancing the modeling of CAR T-cell therapy. This chip incorporates an integrated biosensor for real-time cytokine detection, providing essential insights into cytokine dynamics throughout the therapy process. This advanced chip enables spatiotemporal monitoring of CAR T-cell functionality by observing key processes such as T-cell extravasation, target recognition, and leukemia cell destruction. Additionally, it supports the detailed examination of immune cell interactions, including those with monocytes and basophils, which are critical for understanding the side effects of CAR T therapy, particularly its impact on neuronal tolerance. By meticulously tracking these dynamics, our tool not only refines the precision of therapy modeling but also deepens our understanding of the immune response and potential toxicities associated with CAR T-cell therapy. Our investigations using the chip have yielded detailed profiles of cytokine levels across various therapy phases, including remission, resistance, and relapse. Moreover, the chip is capable of evaluating the effects of compounds in inhibiting cytokine release. This cutting-edge tool significantly shortens the development timeline, reduces costs, and enhances the physiological relevance of preclinical evaluations for CAR T-cell therapy. The implementation of this ‘(pre-)clinical-trial-on-chip’ platform is poised to revolutionize CAR T-cell therapy by facilitating the development of personalized therapies and informing clinical decision-making processes. Our findings represent a transformative step forward in the optimization and safety profiling of CAR T-cell therapies. Citation Format: Xuejia Kang, Lang Zhou, Weiqiang Chen, Zongliang Yue, Pengyu Chen. Development of an Integrated Microfluidic Chip for CAR T-Cell Therapy: A Preclinical Trial-on-Chip Tool [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4612.

Despite the transformative impact of immune checkpoint inhibitors (ICIs) targeting the PD-1/PD-L1 pathway in cancer therapy, up to 80% of patients fail to respond, necessitating reliable predictive biomarkers to guide treatment decisions. Recent studies highlight the critical role of tumor-derived exosomal PD-L1 in immune evasion, and its potential as a diagnostic and prognostic biomarker in cancer immunotherapy. However, significant challenges remain in elucidating the functional roles of PD-L1+ exosomes in immune suppression, as current methods lack the ability to precisely and simultaneously characterize and monitor exosome secretion and the corresponding immune modulation on site. To address this, we developed an integrated microfluidic platform that combines a digital nanoplasmonic immunoassay with a cell-on-a-chip system, enabling in situ monitoring of PD-L1+ exosome secretion and exosome-mediated T cell immune responses. This nanoplasmonic immunoassay integrated cell-on-a-chip (NIIC) creates a localized co-cultured microenvironment that facilitates exosome-mediated cellular interactions without direct contact. The NIIC employs machine-learning assisted signal processing for highly sensitive detection of both exosomes and cytokines, providing spatial and quantitative analysis of immune modulation in situ. Using this system, we demonstrated that PD-L1+ exosomes from cancer cells significantly suppressed IFN-γ and IL-2 secretion in neighboring T cells, offering direct insights into exosome-mediated immune suppression. The NIIC platform represents a powerful tool for advancing the understanding of exosome-driven immune modulation and holds potential for predicting clinical responses to anti-PD-1/PD-L1 therapies, paving the way for more personalized cancer immunotherapy strategies.

The fidelity of immune responses depends on timely controlled and selective mRNA degradation that is largely driven by RNA-binding proteins (RBPs). It remains unclear whether stochastic or directed processes govern the selection of an individual mRNA molecule for degradation. Using human and mouse cells, we show that tristetraprolin (TTP, also known as ZFP36), an essential anti-inflammatory RBP, destabilizes target mRNAs via a hierarchical molecular assembly. The assembly formation strictly relies on the interaction of TTP with RNA. The TTP homolog ZFP36L1 exhibits similar requirements, indicating a broader relevance of this regulatory program. Unexpectedly, the assembly of the cytoplasmic mRNA-destabilization complex is licensed in the nucleus by TTP binding to pre-mRNA, which we identify as the principal TTP target rather than mRNA. Hence, the fate of an inflammation-induced mRNA is decided concomitantly with its synthesis. This mechanism prevents the translation of excessive and potentially harmful inflammation mediators, irrespective of transcription.

Many cancer therapies fail in clinical trials despite showing potent efficacy in preclinical studies. One of the key reasons is the adopted preclinical models cannot recapitulate the complex tumor microenvironment (TME) and reflect the heterogeneity and patient specificity in human cancer. Cancer-on-a-chip (CoC) microphysiological systems can closely mimic the complex anatomical features and microenvironment interactions in an actual tumor, enabling more accurate disease modeling and therapy testing. This review article concisely summarizes and highlights the state-of-the-art progresses in CoC development for modeling critical TME compartments including the tumor vasculature, stromal and immune niche, as well as its applications in therapying screening. Current dilemma in cancer therapy development demonstrates that future preclinical models should reflect patient specific pathophysiology and heterogeneity with high accuracy and enable high-throughput screening for anticancer drug discovery and development. Therefore, CoC should be evolved as well. We explore future directions and discuss the pathway to develop the next generation of CoC models for precision cancer medicine, such as patient-derived chip, organoids-on-a-chip, and multi-organs-on-a-chip with high fidelity. We also discuss how the integration of sensors and microenvironmental control modules can provide a more comprehensive investigation of disease mechanisms and therapies. Next, we outline the roadmap of future standardization and translation of CoC technology toward real-world applications in pharmaceutical development and clinical settings for precision cancer medicine and the practical challenges and ethical concerns. Finally, we overview how applying advanced artificial intelligence tools and computational models could exploit CoC-derived data and augment the analytical ability of CoC.

The self-assembled peptide RADA16-I (RADARADARADARADA) has been widely used in biomaterials. However, studies on the practical application of self-assembled peptide hydrogels loaded with bioactive peptides are still insufficient. In this study, we successfully prepared the peptide nanofiber gel RGJ by incorporating the bioactive peptides A8SGLP-1 (G) and Jagged-1 (J) into RADA16-I (R) in specific ratios. The mechanical properties, secondary structure, and microstructure of RGJ were thoroughly characterized using a rheometer, circular dichroism (CD), and transmission electron microscopy (TEM). The results showed that R and RGJ adopted stable β-folded structures at room temperature, and RGJ exhibited a nanofiber mesh structure, confirming its excellent physical properties. Cellular experiments demonstrated that RGJ significantly enhanced the proliferation and migration of HaCaT, L929, and HUVEC cells, with the most pronounced effect observed in HUVEC cells. In the 100 μg/mL RGJ-treated group, cell viability (OD value) reached 1.369, which was significantly higher than that of the control group (0.673) and the R-only group (0.848). The strongest pro-migratory effect was observed in HaCaT cells, with a scratch closure rate of 22.83 %. In vivo experiments showed that the deep second-degree burn wounds of mice in the RGJ gel-treated group healed rapidly by day 17, exhibiting 99.5 % wound closure, compared to 84.02 % in the R gel group, and 73.02 % and 70.97 % in the control and burn cream groups, respectively. Immunohistochemistry and ELISA results further confirmed that RGJ significantly reduced wound and systemic inflammatory responses while promoting the secretion of pro-angiogenic factors VEGF and CD31, revealing its potential mechanism for enhancing burn wound healing. Additionally, RGJ significantly reduced wound scar formation and increased skin collagen deposition, demonstrating a favorable biosafety profile compared to the control group, commercial burn ointment, and the R-only treatment group. In conclusion, the development of the peptide nanofiber gel RGJ holds great potential for wound management applications and lays a foundation for future related research.

This web-based cross-sectional study aimed to evaluate the knowledge, attitudes, and practices of physicians and hospital pharmacists towards polypharmacy in older adult patients with chronic diseases in China. This study enrolled 374 physicians and pharmacists (270 females, 92 physicians) in 20 Chinese provinces between December 2022 and March 2023. The knowledge, attitude, and practice scores were 12.65 ± 2.05 (possible range, 0-18), 29.07 ± 2.68 (possible range, 7-35), and 26.16 ± 5.56 (possible range, 7-35), respectively. Working as a hospital pharmacist (vs. physician) was independently associated with adequate knowledge (OR = 2.190; 95% CI = 1.291-2.713; P = 0.004). Working in a tertiary hospital (OR = 4.296; 95% CI = 1.390-13.272; P = 0.011) was independently associated with a positive attitude. Knowledge score (OR = 1.176; 95%CI = 1.038-1.333; P = 0.011), hospital pharmacist (OR = 0.276; 95% CI = 0.137-0.557; P < 0.001), master's degree or higher (OR = 1.754; 95% CI = 1.011-3.045; P = 0.046) and senior professional title (OR = 2.020; 95% CI = 1.032-3.952; P = 0.040) were independently associated with proactive practice toward polypharmacy in older adults. Physicians and hospital pharmacists had favorable knowledge, positive attitudes, and proactive practice toward polypharmacy. In conclusion, enhancing knowledge through continuous education, promoting interprofessional collaboration, educating patients, and conducting regular evaluations for quality improvement are necessary to improve the KAP of healthcare professionals toward polypharmacy in older adults.

T cell-based immunotherapy has recently emerged as a promising strategy to treat cancer, requiring the activation of antigen-directed cytotoxicity to eliminate cancer cells. Mechanical signaling, although often overshadowed by its biochemical counterpart, plays a crucial role in T cell anticancer responses, from activation to cytolytic killing. Rapid advancements in the fields of chemistry, biomaterials, and micro/nanoengineering offer an interdisciplinary approach to incorporating mechano- and immunomodulatory ligands, including but not limited to synthetic peptides, small molecules, cytokines, and artificial antigens, onto the biomaterial-based platforms to modulate mechanotransducive processes in T cells. The surface engineering of these immunomodulatory ligands with optimization of ligand density, geometrical arrangement, and mobility has been proven to better mimic the natural ligation between immunoreceptors and ligands to directly enhance or inhibit mechanotransduction pathways in T cells, through triggering upstream mechanosensitive channels, adhesion molecules, cytoskeletal components, or downstream mechanoimmunological regulators. Despite its tremendous potential, current research on this new biomaterial surface engineering approach for mechanomodulatory T cell activation and effector functions remains in a nascent stage. This review highlights the recent progress in this new direction, focusing on achievements in mechanomodulatory ligand-based surface engineering strategies and underlying principles, and outlooks the further research in the rapidly evolving field of T cell mechanotransduction engineering for efficient immunotherapy.

Following improved accessibility to medical services, the phenomenon of polypharmacy in elderly patients with comorbidity has been increasing globally. Polypharmacy patients are prone to drug interactions, adverse drug reactions, and even the risk of death etc. Therefore, there is an urgent need to fully understand the current status and characteristics of drug use in elderly patients with chronic diseases, focusing on polypharmacy factors to ensure that medications for elderly patients are effective and safe. To collect and analyze the characteristics of the current drug use situation in elderly patients with chronic diseases in Chongqing and further explore the influencing factors for polypharmacy, providing references for formulating more effective and safe medication regimens for elderly patients. Most elderly patients affected with chronic diseases in Chongqing were willing to go to hospitals or pharmacies to buy medicines. However, they were not familiar with their disease conditions and drug-related adverse reactions and could not be regularly followed up or monitored. The number of diseases, medications, and adverse drug reactions increased with the increasing age of elderly patients. The problem of irrational use of drugs in elderly patients with chronic diseases was relatively prominent, especially the use of traditional Chinese medicines. The medication situation in elderly patients with chronic diseases was not optimistic, and the problem of polypharmacy was relatively prominent. Further large-scale studies are needed to provide a certain reference for improving the current status of drug use in elderly patients.

Coronavirus disease 2019 (COVID-19) may predispose patients to cardiac injuries but whether COVID-19 infection affects the morphological features of coronary plaques to potentially influence the outcome of patients with coronary artery disease (CAD) remains unknown. By using optical coherence tomography (OCT), this study compared the characteristics of coronary plaque in CAD patients with/without COVID-19 infection. The 206 patients were divided into two groups. The COVID-19 group had 113 patients between December 7, 2022 and March 31, 2023 who received optical coherence tomography (OCT) assessment after China decided to lift the restrict on COVID-19 and had a history of COVID-19 infection. The non-COVID-19 group had 93 patients without COVID-19 infection who underwent OCT before December 7, 2022. The COVID-19 group demonstrated a higher incidence of plaque ruptures (53.1% vs. 38.7%, p=0.039), erosions (28.3% vs. 11.8%, p=0.004), fibrous (96.5% vs. 89.2%, p=0.041) and diffuse lesions (73.5% vs. 50.5%, p<0.001) compared to the non-COVID-19 group, whereas non-COVID-19 group exhibited a higher frequency of cholesterol crystals (83.9% vs. 70.8%, p=0.027), deep calcifications (65.6% vs. 51.3%, p=0.039) and solitary lesions (57.0% vs. 34.5%, p=0.001). Kaplan-Meier survival analysis revealed a significantly lower major adverse cardiac events (MACE)-free probability in COVID-19 group (91.6% vs. 95.5%, P=0.006) than non-COVID-19 group. In conclusion, OCT demonstrated that COVID-19 infection is associated with coronary pathological changes such as more plaque ruptures, erosions, and fibrosis as well as diffuse lesions. Further, COVID-19 infection is associated with the higher propensity for acute coronary events and the higher risk of MACE in CAD patients.

Background: A high proportion of coronary microvascular dysfunction (CMD) has been observed in patients with acute myocardial infarction (AMI) who have received primary percutaneous coronary intervention (PCI), which may affect their prognosis. This study used cadmium zinc telluride (CZT) single photon emission computed tomography (SPECT) to evaluate the prevalence and characteristics of CMD and myocardial area at risk (AAR) in AMI patients who had undergone primary PCI. Methods: We conducted a single-center cross-sectional retrospective study at TEDA International Cardiovascular Hospital from September 2021 to June 2022. A total of 83 patients received primary PCI for AMI. Subsequently, a rest/stress dynamic and routine gated myocardial perfusion imaging (MPI) were performed 1 week after PCI. The CMD group was defined as having a residual stenosis of infarct-related artery (IRA) <50% and myocardial flow reserve (MFR) <2.0 in this corresponding territory, whereas MFR ≥2.0 of IRA pertained to the normal control group. Rest-AAR of infarction (%) and stress-AAR (%) were expressed by the percentage of measured rest-defect-size and stress-defect-size in the left ventricular area, respectively. Logistic regression analyses were performed to identify significant predictors of CMD. Results: A total of 53 patients with a mean age of 57.06±11.99 years were recruited, of whom 81.1% were ST-segment elevation myocardial infarction (STEMI). The proportion of patients with CMD was 79.2% (42/53). The time of pain to SPECT imaging was 7.50±1.27 days in the CMD group and 7.45±1.86 days among controls. CMD patients had a higher body mass index (BMI) than controls (26.48±3.26 vs. 24.36±2.73 kg/m2, P=0.053), and a higher proportion of STEMI, thrombolysis in myocardial infarction (TIMI) 0 grade of IRA prior PCI than controls (88.1% vs. 54.5%, P=0.011; 61.9% vs. 18.2%, P=0.004, respectively). No significant difference was identified in the rest-myocardial blood flow (MBF) of IRA between the 2 groups, whereas the stress-MBF and MFR of IRA, rest-AAR, and stress-AAR in the CMD group were remarkably lowered. Higher BMI [odds ratio (OR): 1.332, 95% confidence interval (CI): 1.008–1.760, P=0.044] and stress-AAR (OR: 1.994, 95% CI: 1.122–3.543, P=0.019) were used as independent predictors of CMD occurrence. Conclusions: The prevalence of CMD is high in AMI patients who received primary PCI. Each 1 kg/m2 increase in BMI was associated with a 1.3-fold increase in CMD risk. A 5% increase in stress-AAR was associated with a nearly 2-fold increase in CMD risk. Increased BMI and stress-AAR predicts decreased coronary reserve function.

Cover legend: The cover image is based on the Research Article Aging-associated decline in vascular smooth muscle cell mechanosensation is mediated by Piezo1 channel by Ngoc Luu et al., https://doi.org/10.1111/acel.14036

Abstract The rise in antibiotic-resistant pathogens, highly infectious viruses, and chronic diseases has prompted the search for rapid and versatile medical tests that can be performed by the patient. An electronic biosensing platform based on field-effect transistors (FETs) is particularly attractive due to sensitivity, fast turn-around, and compatibility with semiconductor manufacturing. However, the lack of methods for pathogen-specific functionalization of individual FETs prevents parallel detection of multiple pathogens. Indeed, so far functionalization of FET based biosensors is achieved by drop casting without any spatial selectivity. Here, we propose a paradigm shift in FET’s biofunctionalization. Specifically, we use thermal scanning probe lithography (tSPL) with a thermochemically sensitive polymer that can be spin-coated on any FET material. We demonstrate that this scalable, CMOS compatible methodology can be used to functionalize individual FETs with different bioreceptors on the same chip, at sub-20 nm resolution, paving the way for massively parallel FET detection of multiple pathogens. Antibody- and aptamer-modified FET sensors are then realized, achieving an ultra-sensitive detection of 5 aM of SARS-CoV-2 spike proteins and 10 human SARS-CoV-2 infectious live virus particles/ml, and selectivity against human influenza A (H1N1) live virus.

Abstract Abstract Body: Characteristics of the PDAC tumor microenvironment (TME) such as desmoplasia and an immunosuppressive landscape contribute to its resistance to chemotherapy and immunotherapy approaches. To recapitulate and dissect the in vivo PDAC TME, we bioengineered a PDAC organoid-on-a-chip model by integrating patient-derived PDAC organoids on a microfluidic-based organ-on-a-chip. While the patient-derived organoids (PDOs) maintain the characteristics of the in vivo counterparts with high fidelity, the organ-on-a-chip provides a controllable and reproducible environment with stromal and immune niche cells, creating an in vitro model that may providing insights into PDAC drug resistance and allow testing for new therapies. The multicellular organoid-on-a-chip model is composed of 1) a central PDAC niche including patient-derived PDAC organoids, patient-derived cancer-associated macrophages (CAFs), tumor-associated macrophages (TAMs) and blood vessels, and 2) surrounding vascular networks for nutrient transport and drug delivery. We reproduced the hypovasculariy of PDAC stromal niche on the chip and found a lower level of CD31 expression in the PDAC organoid niche than in the normal pancreatic organoid niche (p&amp;lt;0.0001). Comparing with the niche without PDOs, the PDAC niche on chip demonstrated a higher expression level of α-smooth muscle actin (α-SMA) on CAFs (fold-change=2.32, p&amp;lt;0.0001) and a more extensive deposition of ECM components (e.g., collagen I, collagen III, collagen IV and hyaluronic acid, fold-change&amp;gt;=2.25, p⇐0.0002), indicating the existence of desmoplasia on chip. Profiling of cytokine levels in the media revealed that, compared to models used with normal pancreatic epithelial cells with the other cellular components, the PDAC organoid TME induced the upregulation of a series of pro-tumor cytokines including MIP-3α, IL-8, CXCL5, IL-13 (fold-change&amp;gt;=5.02), the inflammation-related cytokines MIF, IL-5, MCP-1 (fold-change&amp;gt;=1.82), and CAF-derived cytokines like CXCL10, HGF, CXCL1, CXCL12 (fold-change&amp;gt;=1.82). The immunosuppressive cytokine TGF-β was increased after PDOs were loaded on the chip (762.3 pg/mL vs. 222.6 pg/mL in conditioned medium), as was the proinflammatory cytokine IL-6 (1089.5 pg/mL vs. 81.3 pg/mL), was maintained at a higher level than that in the niche with normal organoids (380.5 pg/mL vs. 60.4 pg/mL at day 13). We also found that PDOs induced TAMs inclined to be M2-like (CD68+CD163+) phenotype on chip. In turn niche cells (TAMs and CAFs) contributed to the immunosuppression in PDAC TME: the secretion of cytokines like IL-6, IL-13, MIP-1β and HGF were downregulated without niche cells (fold-change⇐0.646). In conclusion, we have developed a PDAC organoid-on-a-chip recapitulates the PDAC TME and is promising to serving as a platform to study mechanisms of drug resistance in PDAC as well as test novel therapies. Citation Format: Lunan Liu, Diane M. Simeone, Weiqiang Chen. An organoids-on-a-chip model to recapitulate and dissect the tumor microenvironment of PDAC [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B057.

The rise in antibiotic-resistant pathogens, highly infectious viruses, and chronic diseases has prompted the search for rapid and versatile medical tests that can be performed by the patient. Field-effect transistor (FET)-based electronic biosensing platforms are particularly attractive due to their sensitivity, fast turn-around time, potential for parallel detection of multiple pathogens, and compatibility with semiconductor manufacturing. However, an unmet critical need is a scalable, site-selective multiplexed biofunctionalization method with nanoscale precision for immobilizing different types of pathogen-specific bioreceptors on individual FETs, preventing parallel detection of multiple targets. Here, we propose a paradigm shift in FET biofunctionalization using thermal scanning probe lithography (tSPL) with a thermochemically sensitive polymer. This polymer can be spin-coated on fully-fabricated FET chips, making this approach applicable to any FET sensor material and technology. Crucially, we demonstrate the spatially selective multiplexed functionalization capability of this method by immobilizing different types of bioreceptors at prescribed locations on a chip with sub-20 nm resolution, paving the way for massively parallel FET detection of multiple pathogens. Antibody- and aptamer-modified graphene FET sensors are then realized, achieving ultra-sensitive detection of a minimum measured concentrations of 3 aM of SARS-CoV-2 spike proteins and 10 human SARS-CoV-2 infectious live virus particles per ml, and selectivity against human influenza A (H1N1) live virus.

ABSTRACT The fidelity of immune responses is dependent on a timely controlled and selective mRNA degradation that is largely driven by RNA-binding proteins (RBPs). It remains unclear whether the selection of an individual mRNA molecule for degradation is governed by stochastic or directed processes. Here, we show that tristetraprolin (TTP, also known as ZFP36), an essential anti-inflammatory RBP, destabilized the target mRNA via a hierarchical molecular assembly. The assembly formation is strictly reliant on TTP interaction with RNA. The TTP homolog ZFP36L1 exhibits similar requirements indicating a broader relevance of this regulatory program. Unexpectedly, the assembly of the cytoplasmic mRNA-destabilization complex is licensed in the nucleus by TTP binding to pre-mRNA while mature cytoplasmic mRNA does not constitute a de novo TTP target. Hence, the fate of an inflammation-induced mRNA is decided concomitantly with its synthesis. This fate decision mechanism prevents the translation of superfluous and potentially harmful inflammation mediators, and ensures an efficient cessation of the immune response irrespective of transcriptional activity.

Aging of the vasculature is associated with detrimental changes in vascular smooth muscle cell (VSMC) mechanosensitivity to extrinsic forces in their surrounding microenvironment. However, how chronological aging alters VSMCs' ability to sense and adapt to mechanical perturbations remains unexplored. Here, we show defective VSMC mechanosensation in aging measured with ultrasound tweezers-based micromechanical system, force instantaneous frequency spectrum, and transcriptome analyses. The study reveals that aged VSMCs adapt to a relatively inert mechanobiological state with altered actin cytoskeletal integrity, resulting in an impairment in their mechanosensitivity and dynamic mechanoresponse to mechanical perturbations. The aging-associated decline in mechanosensation behaviors is mediated by hyperactivity of Piezo1-dependent calcium signaling. Inhibition of Piezo1 alleviates vascular aging and partially restores the loss in dynamic contractile properties in aged cells. Altogether, our study reveals the signaling pathway underlying aging-associated aberrant mechanosensation in VSMC and identifies Piezo1 as a potential therapeutic mechanobiological target to alleviate vascular aging.

The mechanical constraints in the overcrowding glioblastoma (GBM) microenvironment have been implicated in the regulation of tumor heterogeneity and disease progression. Especially, such mechanical cues can alter cellular DNA transcription and give rise to a subpopulation of tumor cells called cancer stem cells (CSCs). These CSCs with stem-like properties are critical drivers of tumorigenesis, metastasis, and treatment resistance. Yet, the biophysical and molecular machinery underlying the emergence of CSCs in tumor remained unexplored. This work employed a two-dimensional micropatterned multicellular model to examine the impact of mechanical constraints arisen from geometric confinement on the emergence and spatial patterning of CSCs in GBM tumor. Our study identified distinct spatial distributions of GBM CSCs in different geometric patterns, where CSCs mostly emerged in the peripheral regions. The spatial pattern of CSCs was found to correspond to the gradients of mechanical stresses resulted from the interplay between the cell-ECM and cell-cell interactions within the confined environment. Further mechanistic study highlighted a Piezo1-RhoA-focal adhesion signaling axis in regulating GBM cell mechanosensing and the subsequent CSC phenotypic transformation. These findings provide new insights into the biophysical origin of the unique spatial pattern of CSCs in GBM tumor and offer potential avenues for targeted therapeutic interventions.

Immunomodulation therapies have attracted immense interest recently for the treatment of immune-related diseases, such as cancer and viral infections. This new wave of enthusiasm for immunomodulators, predominantly revolving around cytokines, has spurred emerging needs and opportunities for novel immune monitoring and diagnostic tools. Considering the highly dynamic immune status and limited window for therapeutic intervention, precise real-time detection of cytokines is critical to effectively monitor and manage the immune system and optimize the therapeutic outcome. The clinical success of such a rapid, sensitive, multiplex immunoanalytical platform further requires the system to have ease of integration and fabrication for sample sparing and large-scale production toward massive parallel analysis. In this article, we developed a nanoplasmonic bioink-based, label-free, multiplex immunosensor that can be readily "written" onto a glass substrate via one-step calligraphy patterning. This facile nanolithography technique allows programmable patterning of a minimum of 3 μL of nanoplasmonic bioink in 1 min and thus enables fabrication of a nanoplasmonic microarray immunosensor with 2 h simple incubation. The developed immunosensor was successfully applied for real-time, parallel detection of multiple cytokines (e.g., interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and transforming growth factor-beta (TGF-β)) in immunomodulated macrophage samples. This integrated platform synergistically incorporates the concepts of nanosynthesis, nanofabrication, and nanobiosensing, showing great potential in the scalable production of label-free multiplex immunosensing devices with superior analytical performance for clinical applications in immunodiagnostics and immunotherapy.

Metasurfaces, composed of micro-nano-structured planar materials, offer highly tunable control over incident light and find significant applications in imaging, navigation, and sensing. However, highly efficient polarization devices are scarce for the extended shortwave infrared (ESWIR) range (1.7~2.5 μm). This paper proposes and demonstrates a highly efficient all-dielectric diatomic metasurface composed of single-crystalline Si nanocylinders and nanocubes on SiO2. This metasurface can serve as a nanoscale linear polarizer for generating polarization-angle-controllable linearly polarized light. At the wavelength of 2172 nm, the maximum transmission efficiency, extinction ratio, and linear polarization degree can reach 93.43%, 45.06 dB, and 0.9973, respectively. Moreover, a nonpolarizing beam splitter (NPBS) was designed and deduced theoretically based on this polarizer, which can achieve a splitting angle of ±13.18° and a phase difference of π. This beam splitter can be equivalently represented as an integration of a linear polarizer with controllable polarization angles and an NPBS with one-bit phase modulation. It is envisaged that through further design optimization, the phase tuning range of the metasurface can be expanded, allowing for the extension of the operational wavelength into the mid-wave infrared range, and the splitting angle is adjustable. Moreover, it can be utilized for integrated polarization detectors and be a potential application for optical digital encoding metasurfaces.

Microgravity accelerates the aging of various physiological systems, and it is well acknowledged that aged individuals and astronauts both have increased susceptibility to infections and poor response to vaccination. Immunologically, dendritic cells (DCs) are the key players in linking innate and adaptive immune responses. Their distinct and optimized differentiation and maturation phases play a critical role in presenting antigens and mounting effective lymphocyte responses for long-term immunity. Despite their importance, no studies to date have effectively investigated the effects of microgravity on DCs in their native microenvironment, which is primarily located within tissues. Here, we address a significantly outstanding research gap by examining the effects of simulated microgravity via a random positioning machine on both immature and mature DCs cultured in biomimetic collagen hydrogels, a surrogate for tissue matrices. Furthermore, we explored the effects of loose and dense tissues via differences in collagen concentration. Under these various environmental conditions, the DC phenotype was characterized using surface markers, cytokines, function, and transcriptomic profiles. Our data indicate that aged or loose tissue and exposure to RPM-induced simulated microgravity both independently alter the immunogenicity of immature and mature DCs. Interestingly, cells cultured in denser matrices experience fewer effects of simulated microgravity at the transcriptome level. Our findings are a step forward to better facilitate healthier future space travel and enhance our understanding of the aging immune system on Earth.

Plant ATP-binding cassette (ABC) transporters contribute the transport of diverse secondary metabolites. However, their roles in cannabinoid trafficking are still unsolved in Cannabis sativa. In this study, 113 ABC transporters were identified and characterized in C. sativa from their physicochemical properties, gene structure, and phylogenic relationship, as well as spatial gene expression patterns. Eventually, seven core transporters were proposed including one member in ABC subfamily B (CsABCB8) and six ABCG members (CsABCG4, CsABCG10, CsABCG11, CsABCG32, CsABCG37, and CsABCG41), harboring potential in participating cannabinoid transport, by combining phylogenetic and co-expression analysis from the gene and metabolite level. The candidate genes exhibited a high correlation with cannabinoid biosynthetic pathway genes and the cannabinoid content, and they were highly expressed where cannabinoids appropriately biosynthesized and accumulated. The findings underpin further research on the function of ABC transporters in C. sativa, especially in unveiling the mechanisms of cannabinoid transport to boost systematic and targeted metabolic engineering.

Aging of the vasculature is associated with detrimental changes in vascular smooth muscle cell (VSMC) mechanosensitivity to extrinsic forces in their surrounding microenvironment. However, how chronological aging alters VSMCs' ability to sense and adapt to mechanical perturbations remains unexplored. Here, we show defective VSMC mechanosensation in aging measured with ultrasound tweezers-based micromechanical system, force instantaneous frequency spectrum and transcriptome analyses. The mechanobiological study reveals that aged VSMCs adapt a relatively inert solid-like state with altered actin cytoskeletal integrity, resulting in an impairment in their mechanosensitivity and dynamic mechanoresponse to mechanical perturbations. The aging-associated decline in mechanosensation behaviors is mediated by hyperactivity of Piezo1-dependent calcium signaling. Inhibition of Piezo1 alleviates vascular aging and partially restores the loss in dynamic contractile properties in aged cells. Altogether, our study reveals the novel signaling pathway underlying aging-associated aberrant mechanosensation in VSMC and identifies Piezo1 as a potential therapeutic mechanobiological target to alleviate vascular aging.

Chimeric antigen receptor (CAR) T cell immunotherapy is promising for treatment of blood cancers; however, clinical benefits remain unpredictable, necessitating development of optimal CAR T cell products. Unfortunately, current preclinical evaluation platforms are inadequate due to their limited physiological relevance to humans. We herein engineered an organotypic immunocompetent chip that recapitulates microarchitectural and pathophysiological characteristics of human leukemia bone marrow stromal and immune niches for CAR T cell therapy modeling. This leukemia chip empowered real-time spatiotemporal monitoring of CAR T cell functionality, including T cell extravasation, recognition of leukemia, immune activation, cytotoxicity, and killing. We next on-chip modelled and mapped different responses post CAR T cell therapy, i.e., remission, resistance, and relapse as observed clinically and identify factors that potentially drive therapeutic failure. Finally, we developed a matrix-based analytical and integrative index to demarcate functional performance of CAR T cells with different CAR designs and generations produced from healthy donors and patients. Together, our chip introduces an enabling ‘(pre-)clinical-trial-on-chip’ tool for CAR T cell development, which may translate to personalized therapies and improved clinical decision-making.