Cell Activity Research Articles

Spatially Quantitative Imaging of Enzyme Activity in a Living Cell.

Enzyme activity plays a key role in cell heterogeneity. Its spatially quantitative imaging in a living cell not only directly displays but also helps people to understand cell heterogeneity. Current methods are hard to achieve due to the short intracellular retention or lack of internal reference of the imaging probes. Herein, we rationally designed a self-referenced Raman probe Val-Cit-Cys(StBu)-Pra-Gly-CBT (Yne-CBT) which takes an intracellular cathepsin B (CTSB)-initiated CBT-Cys click reaction to yield a long-retained cyclic dimer in cell. In the meantime, Raman signal changes of its two chemical bonds (C≡C and C≡N) after the reaction are used for self-referencing and quantitative Raman imaging of CTSB activity. In vitro experiments demonstrated that, with shell-isolated nanoparticle-enhanced Raman spectroscopy technique, 20 μM Yne-CBT was able to quantitatively detect CTSB activity with a limit of detection of 61.4 U L-1. Under a homemade microfluidic channel, Yne-CBT was successfully applied for spatially quantitative imaging CTSB activity in a living cell. Our strategy provides people with a facile method to directly and quantitatively display cell heterogeneity.

Investigating the mechanism of auxin-mediated fulvic acid-regulated root growth in Oryza sativa through physiological and transcriptomic analyses.

As rice is one of the most crucial staple food sources worldwide, enhancing rice yield is paramount for ensuring global food security. Fulvic acid (FA), serving as a plant growth promoter and organic fertilizer, holds significant practical importance in studying its impact on rice root growth for improving rice yield and quality. This study investigated the effects of different concentrations of FA on the growth of rice seedlings. The results indicated that 0.05 g/L FA could promote the growth of rice seedlings, while 0.5 g/L FA inhibited root growth, reduced cell activity and enzyme activity in the root tips, and accumulated reactive oxygen species in root cells. To further elucidate the molecular mechanisms underlying these effects, we performed transcriptomic analysis and found that auxin (Aux) may be involved in the growth process mediated by FA. Furthermore, transcriptome heatmap analysis revealed a significant upregulation of the Aux/indoleacetic acid (Aux/IAA) gene family after FA treatment, suggesting that this gene family plays a crucial role in the impact of FA on root growth. Additionally, by detecting endogenous Aux content and adding exogenous Aux inhibitors, we confirmed the involvement of FA in rice seedling root growth as well as in the synthesis and transduction pathway of Aux.

Acetyltransferase NAT10 promotes an immunosuppressive microenvironment by modulating CD8+ T cell activity in prostate cancer

N-acetyltransferase 10 (NAT10), an enzyme responsible for ac4C acetylation, is implicated in cancer progression, though its specific biological function in prostate cancer remains insufficiently understood. This study clarifies NAT10’s role in prostate cancer and its effects on the tumor immune microenvironment. NAT10 expression and clinical relevance were assessed through bioinformatics, RT-qPCR, and IHC analyses, comparing prostate cancer tissues with normal controls. The impact of NAT10 on tumor cell proliferation, migration, and invasion was investigated via in vitro assays—including CCK-8, EdU, wound healing, and 3D-Transwell—as well as in vivo mouse xenograft models and organoid studies. Further, NAT10’s influence on immune cell infiltration was examined using flow cytometry, IHC, cell co-culture assays, and ELISA to elucidate downstream chemokine effects, specifically targeting CD8+ T cells. Findings indicated significant upregulation of NAT10 in prostate cancer cells, enhancing their proliferative and invasive capacities. Notably, NAT10 suppresses CD8+ T cell recruitment and cytotoxicity through the CCL25/CCR9 axis, fostering an immunosuppressive microenvironment that exacerbates tumor progression. An ac4C modification score was also devised based on NAT10’s downstream targets, providing a novel predictive tool for evaluating immune infiltration and forecasting immunotherapy responses in patients with prostate cancer. This study underscores NAT10’s pivotal role in modulating the prostate cancer immune microenvironment, offering insights into the immune desert phenomenon and identifying NAT10 as a promising therapeutic target for improving immunotherapy efficacy.

Abstract PR001: Discovery of Etrumadenant, a first-in-class dual A2a and A2b adenosine receptor antagonist for cancer immunotherapy

Abstract High levels of adenosine are generated in the tumor microenvironment (TME) by sequential hydrolysis of extracellular ATP by the ecto-nucleotidases CD39 (ATP→AMP) and CD73 (AMP→adenosine). Adenosine activates A2a and A2b adenosine receptors on immune cells, resulting in immunosuppression. Since A2aR is expressed by a variety of lymphocytes, the suppressive effects of adenosine on this cell type can be potentially reversed by blocking the A2aR. In contrast, myeloid cells that have comparable expression of A2aR and A2bR require dual A2a/bR blockade for full reversal of the immunosuppressive effects of adenosine. The first generation of adenosine antagonists deployed in oncology were repurposed CNS drug candidates with properties that were deemed less than optimal for anti-tumor efficacy (e.g., A2aR-selective, good BBB permeability, not optimized for minimal plasma protein binding, etc.). We sought to design a potent and selective dual antagonist of A2aR and A2bR to effectively block the high concentrations of adenosine found in tumors, with diminished brain penetration, and minimal non-specific binding to plasma proteins. Using a pharmacophore mapping approach, we discovered potent A2aR antagonists with favorable PK profile in rats, good selectivity, and moderate potency against A2bR. Systematic SAR studies led to improvement in A2bR potency, which ultimately resulted in Etrumadenant (AB928). Etrumadenant inhibits both A2aR and A2bR with similar potencies (KB: 1.4 nM and 2 nM, respectively), and is highly selective against other targets in the adenosine pathway with minimal penetration across the blood brain barrier. Etrumadenant provides superior dendritic cell activity compared to A2aR antagonist alone. Its combination with immunogenic chemotherapy enhances immune activity and suppresses tumor growth and metastasis in vivo. These data support our rational design of a dual A2a and A2b adenosine receptor antagonist to impact adenosine biology on multiple cell types within the TME. Etrumadenant is currently undergoing phase 2 clinical trials in colorectal and lung cancer patients. Citation Format: Ehesan U. Sharif, Dillon H. Miles, Brandon R. Rosen, Joel Beatty, Jenna L. Jeffrey, Laurent P. P. Debien, Rhiannon Thomas-Tran, Debashis Mandal, Dan Direnzo, Sachie Marubayashi, Kristen Zhang, Ferdie Soriano, Elaine Ginn, Divyank Soni, Pei-Yu Chen, Lixia Jin, Steve W. Young, Matt J. Walters, Manmohan R. Leleti, Jay P. Powers. Discovery of Etrumadenant, a first-in-class dual A2a and A2b adenosine receptor antagonist for cancer immunotherapy. [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Optimizing Therapeutic Efficacy and Tolerability through Cancer Chemistry; 2024 Dec 9-11; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(12_Suppl):Abstract nr PR001

Systems Engineering Approach Towards Sensitive Cellular Fluorine-19 MRI.

In vivo fluorine-19 MRI using F-based tracer media has shown utility and versatility for a wide range of biomedical uses, particularly immune and stem cell detection, as well as biosensing. As with many advanced MRI acquisition techniques, the sensitivity and limit of detection (LOD) invivo is a key consideration for a successful study outcome. In this review, we analyze the primary factors that limit cell LOD. The achievable sensitivity is strongly dependent on the specific composition of tracer, cell type of interest, cell activity, data acquisition and reconstruction methods, and MRI hardware design. Recent innovations in molecular 19F tracer design and image acquisition-reconstruction methods have achieved significant leaps in 19F MRI sensitivity, and integration of these new materials and methods into studies can result in > 10-fold improvement in LOD. These developments will help unlock the full potential of clinical 19F MRI for biomedical applications.

Histomorphometric analysis of external apical root resorption using L-loops and different intrusive forces

ABSTRACT Purpose This study aimed to conduct a histomorphometric analysis on the impact of varying intrusive forces on external apical root resorption (EARR) in rat molars, using an L-loop was employed to apply intrusion forces, artificially inducing EARR in the maxillary left first molar (M1) of rats. Materials and methods The L-loop was bent on a plaster rat model using stainless steel round wires with diameters of 0.014 and 0.016 inches. Using a universal test force measuring device, L-loops were calibrated to 4 N/mm and 6 N/mm for the 0.014-inch and 0.016-inch diameter stainless steel round wires, respectively. Fifteen-week-old Sprague-Dawley (SD) male rats were used and divided into the control (no loop) and experimental (0.014 and 0.016) groups (n = 5 for each group). The L-loop was attached to the left maxillary molar, applying an intrusive force to M1. After 15 days, the rats were euthanized. Maxillary bones were analysed using micro-computed tomography (μCT), evaluating M1 intrusive distance of vertical tooth movement and counting the number of tartrate-resistant acid phosphatase (TRAP)-positive cells on the surface of the mesial root apex and alveolar bone. Results The L-loop groups exhibited significantly increased M1 intrusive distance and TRAP-positive cell counts compared to controls. Specifically, the 0.016 group demonstrated greater M1 intrusive distance and more TRAP-positive cells than the 0.014 group. The M1 intrusive distance and TRAP-positive cells increased when intrusive forces were applied using L-loops of different diameters. Conclusion Greater intrusive forces enhance TRAP-positive cell activity, influencing EARR and vertical tooth movement.

Enhancement of the Degradation of Phytosterol Side Chains in Mycolicibacterium by Eliminating the Redox Sensitivity of Key Thiolase and Augmenting Cell Activity

Androstenedione (AD) is a vital intermediate in the synthesis of steroid drugs, making its efficient production critical in the steroid drug industry. Acetyl-CoA acetyltransferase (FadA5), a thiolase enzyme, plays an important role in the metabolic process of degrading phytosterol side chains in Mycolicibacterium to produce AD. This work is the first systematic analysis of the role of FadA5 in the transformation of phytosterols by Mycolicibacterium to produce AD. The relationship between redox potential and AD production was examined using resting cells, and it was confirmed that FadA5 is a key enzyme for AD production. Mutating the 87th cysteine of FadA5 to alanine reduced its redox effect, enhancing the substrate tolerance and biotransformation capacity of the strain. Co-expressing Vitreoscilla hemoglobin (VHb) and propionyl-CoA metabolized the transcription activator (PrpR), decreased intracellular reactive oxygen species levels, and improved cell viability. The AD yield of MSP-fA5C87A-VP/ΔfA5 was 2.541 g/L, an increase of 16.83% over the control strain. Using a repeated batch fermentation process, the production efficiency of the MSP-fA5C87A-VP/ΔfA5 strain was 0.658 g/L/d, which was 1.82 times higher than that of the control strain. These findings provide a theoretical basis for understanding and regulating steroid side-chain catabolism in Mycolicibacterium and offer support for the rational modification of industrial strains for steroidal drug precursor production.

CAR T Cell Nanosymbionts: Revealing the Boundless Potential of a New Dyad

Cancer treatment has traditionally focused on eliminating tumor cells but faces challenges such as resistance and toxicity. A promising direction involves targeting the tumor microenvironment using CAR T cell immunotherapy, which has shown potential for treating relapsed and refractory cancers but is limited by high costs, resistance, and toxicity, especially in solid tumors. The integration of nanotechnology into ICAM cell therapy, a concept we have named “CAR T nanosymbiosis”, offers new opportunities to overcome these challenges. Nanomaterials can enhance CAR T cell delivery, manufacturing, activity modulation, and targeting of the tumor microenvironment, providing better control and precision. This approach aims to improve the efficacy of CAR T cells against solid tumors, reduce associated toxicities, and ultimately enhance patient outcomes. Several studies have shown promising results, and developing this therapy further is essential for increasing its accessibility and effectiveness. Our “addition by subtraction model” synthesizes these multifaceted elements into a unified strategy to advance cancer treatment paradigms.

Natural killer cells as promising candidates in Cancer therapeutics and related immune resistance in tumor microenvironment

Natural killer (NK) cells, as critical components of the innate immune system, have emerged as promising candidates in cancer therapy due to their ability to recognize and destroy tumor cells without prior sensitization. Their mechanisms of action, including perforin-granzyme release and antibody-dependent cellular cytotoxicity (ADCC), position them as versatile agents in cancer immunotherapy. Recent advancements, such as chimeric antigen receptor (CAR)-NK cells and cytokine-based stimulation, have demonstrated enhanced efficacy and reduced side effects compared to conventional immunotherapies. However, the immunosuppressive tumor microenvironment (TME) remains a significant obstacle, imposing challenges like immune checkpoint activation, cytokine suppression, and metabolic constraints that impair NK cell activity. This paper explores the therapeutic potential of NK cells, the challenges of immune resistance in the TME, and emerging strategies to enhance NK cell efficacy. Addressing these challenges is crucial for optimizing NK cell-based treatments and achieving durable responses in cancer patients.

KDM1A epigenetically enhances RAD51 expression to suppress the STING-associated anti-tumor immunity in esophageal squamous cell carcinoma

Histone lysine demethylase LSD1, also known as KDM1A, has been found to regulate multiple cancer hallmarks since it was first identified in 2004. Recently, it has emerged as a promising target for stimulating anti-tumor immunity, specifically boosting T cell activity. However, it remains unclear whether and how it remodels the tumor microenvironment to drive oncogenic processes in esophageal squamous cell carcinoma (ESCC). In this study, protein levels in ESCC tissues were evaluated by immunostaining of tissue microarrays. Cell growth was assessed by colony formation assays in vitro and subcutaneous xenograft models in vivo. High-throughput transcriptomics and spatial immune proteomics were performed using bulk RNA sequencing and digital spatial profiling techniques, respectively. Epigenetic regulation of RAD51 by methylated histone proteins was analyzed using chromatin immunoprecipitated quantitative PCR assays. Finally, our clinical data indicate that KDM1A precisely predicts the overall survival of patients with early-stage ESCC. Inhibition of KDM1A blocked the growth of ESCC cells in vitro and in vivo. Mechanistically, our transcriptomics and spatial immune proteomics data, together with rescue assays, demonstrated that KDM1A specifically removes methyl residues from the histone protein H3K9me2, a transcription repressive marker, thus reducing its enrichment at the promoter of RAD51 to epigenetically reactivate its transcription. Additionally, it significantly inhibits the expression of NF-κB signaling-dependent proinflammatory genes IL-6 and IL-1B through RAD51, thus blocking the STING-associated anti-tumor immunity in stromal tumor-infiltrating lymphocytes (sTIL). Overall, our findings not only indicate that KDM1A is a promising target for ESCC patients at early stages but also provide novel mechanistic insights into its spatial regulation of STING-associated anti-tumor immunity in sTILs to drive the oncogenic processes in ESCC. The translation of these findings will ultimately guide more appropriate combinations of spatial immunotherapies with KDM1A inhibitors to improve the overall survival of specific subgroups in ESCC.

On Multicell-Interaction Chip: In Situ Observing the Interactions between the Astrocytes with Lysosomal Dysfunction and BBB Cells.

Lysosomes in astrocytes play vital roles in toxic protein degradation in the brain. Lysosomal dysfunction can lead to abnormal protein deposits, which further induce damage to neurons and the blood-brain barrier (BBB), and thereby affect the interaction between the nervous and vascular systems. Therefore, investigating the interactions between astrocytes with lysosomal dysfunction and BBB cells is of significant importance. However, the lack of effective in vitro models hinders the study of this complex system. Herein, an 8-well arrayed microfence multicell interculture chip (AMMIC) with a hydrophilically optimized surface is introduced for investigating the interactions between astrocytes and BBB cells. Then, a novel lysosome-targeted photosensitizer, IVQ-2Br, is synthesized for inducing controllable oxidative stress damage in the lysosomes of astrocytes. By the combination of the 8-well AMMIC and IVQ-2Br, a model for studying the interactions between astrocytes with lysosomal dysfunction and BBB cells has been constructed. Particularly, severe secondary injuries to BBB cells brought about by oxidative stress, including alterations in cell morphology and activity as well as notable DNA damage, are in situ observed on the 8-well AMMIC. The mediators involved in this oxidative stress injury-mediated intercellular communication are validated to be reactive oxygen species (ROS) and exosomes. This work not only presents an in vitro modeling method for studying cell-cell interactions but also demonstrates the potential of in vitro models constructed through the integration of complex microfluidic chip techniques and photosensitizers for advancing biomedical research.

Human colonic organoids for understanding early events of familial adenomatous polyposis pathogenesis.

Patients with familial adenomatous polyposis (FAP) harbor mutations in the APC gene and will develop adenoma and early colorectal cancer. There is no validated treatment, and animal models are not sufficient to study FAP. Our aim was to investigate the early events associated with FAP using the intestinal organoid model in a single-center study using biopsies from nonadenomatous and adenomatous colonic mucosa of FAP patients and from healthy controls (HCs). We analyzed intestinal stem cell (ISC) activity and regulation through organoid development and expression of mRNA and proteins, as well as within colonic crypts. We used several compounds to regulate the signaling pathways controlling ISCs, such as WNT, EGFR, PI3K-AKT, TGF-β, yes-associated protein (YAP), and protease-activated receptors. In addition to their high proliferative capacity, nonadenomatous and adenomatous organoids were characterized by cysts and cysts with buds, respectively, suggesting abnormal maturation. Adenomatous organoids were enriched in the stem cell marker LGR5 and dependent on EGF and TGF-β for their growth. Downstream of EGFR, AKT, β-catenin, and YAP were found to be activated in the adenomatous organoids. While the p110β isoform of PI3K was predominant in adenomatous organoids and essential for their growth, p110α was associated with the immature state of nonadenomatous organoids. We conclude that organoids offer a relevant model for studying FAP, and this work highlights abnormal behaviors of immature cells in both nonadenomatous and adenomatous mucosa of FAP patients, which could be targeted therapeutically. © 2024 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

The Causal Impact of Immune Cell Dynamics on liver failure: An Investigation via Mendelian Randomization (MR) Analysis

Background: Liver failure is a critical clinical condition with complex pathophysiology, where the role of immune cells has been increasingly recognized. However, establishing causality between immune cell activity and liver failure has been challenging. We aimed to investigate the causal role of specific immune cell populations in the development of liver failure using Mendelian randomization (MR) techniques. Methods: Genetic data from the Genome-wide association study (GWAS) database, focusing on 731 immunophenotypes and a liver failure dataset was utilised. Single nucleotide polymorphisms (SNPs) associated with immune cell exposures were identified and used as instrumental variables. MR analyses were performed using the TwoSampleMR package in R, employing various MR techniques, including weighted median, MR Egger, inverse-variance weighted (IVW), and others. The robustness of the MR findings was assessed through heterogeneity, pleiotropy, and leave-one-out sensitivity analyses. Results: Our MR study revealed significant causal associations between several immune cell populations and liver failure. Notably, genetic variants associated with CD28 on CD39+ CD4+ T cells and CD3 on CD39+ secreting T Reg cells were found to be causally linked to an increased risk of liver failure. The IVW MR method, which has been previously validated for its performance, provided the most consistent results across immune cell exposures. Conclusion: Our findings suggest a direct causal role for specific immune cells in liver failure, offering potential targets for future therapeutic strategies.

Modular (universal) CAR-T platforms in vivo: a comprehensive systematic review

BackgroundModular (universal) CAR T-platforms were developed to combat the limitations of traditional CAR-T therapy, allowing for multiple targeting of tumor-associated antigens and the ability to control CAR-T cell activity. The modular CAR-T platform consists of a universal receptor (signaling module) that recognizes an adapter molecule on the soluble module, which is responsible for antigen recognition. Multiple platforms have been developed over the last 12 years, and some of them have entered the clinical trial phase. This systematic review seeks to evaluate the different parameters of modular CAR-T platforms performance in animal models.MethodsA systematic search of literature in the PubMed database and in Google Scholar and BASE (Bielefeld Academic Search Engine) search engines was performed according to predefined eligibility criteria. All studies conducted on xenograft mouse models with any variant of modular CAR-T platforms were included. Forest plots were generated for visual presentation of the extracted quantitative findings (standardized mean difference (SMD) and median survival rate (MSR)).ResultsA total of 33 studies employing 15 different modular CAR-T platforms were included. The platforms varied in terms of CAR-T cells, soluble module doses, and their frequency of administration. The studies showed a reduction in tumor burden and in tumor volume compared to the combined negative group. In comparison with the positive control group, there was no significant change in tumor burden or volume. In all the included studies the experimental group had a higher survival probability compared to the combined negative group at the study endpoint, with no significant difference in survival rate compared to the positive control group.ConclusionThe modular CAR-T platforms are generally effective and are a valuable addition to the arsenal of CAR therapy.Systematic Review Registrationhttps://www.crd.york.ac.uk/prospero/ PROSPERO, identifier CRD42023443984.

From nature to clinic: Quercetin’s role in breast cancer immunomodulation

Immunotherapy has brought hope to many breast cancer patients, but not all patients benefit from it. Quercetin (Qu), a natural product found in various sources, has anti-inflammatory and anti-tumor properties. We conducted a review of the pharmacological research of Qu in regulating anti-tumor immunity in vivo and in vitro. Qu can directly regulate the local tumor microenvironment (TME) by enhancing the activity of immune cells which includes promoting the infiltration of T cells and natural killer (NK) cells, inhibiting the recruitment of myeloid-derived suppressor cells and tumor-associated macrophages. Additionally, Qu inhibits anaerobic glycolysis in tumor cells, thereby reducing the production and transport of lactic acid. It also suppresses tumor angiogenesis by targeting the vascular endothelial growth factor (VEGF) pathway and the vitamin D pathway. Furthermore, Qu can enhance the efficacy of immunotherapy for breast cancer by modulating the systemic microenvironment. This includes inhibiting obesity-related chronic inflammation to decrease the production of inflammatory factors, regulating the composition of intestinal microbiota, and intervening in the metabolism of intestinal flora. At the same time, we also address challenges in the clinical application of Qu, such as low absorption rates and unknown effective doses. In conclusion, we highlight Qu as a natural immunomodulator that enhances immune cell activity and has the potential to be developed as an adjunct for breast cancer.

Inhibition of CD226 co-stimulation suppresses diabetes development in the NOD mouse by augmenting regulatory T cells and diminishing effector T cell function.

Immunotherapeutics targeting T cells are crucial for inhibiting autoimmune disease progression proximal to disease onset in type 1 diabetes. There is an outstanding need to augment the durability and effectiveness of T cell targeting therapies by directly restraining proinflammatory T cell subsets, while simultaneously augmenting regulatory T cell (Treg) activity. Here, we present a novel strategy for preventing diabetes incidence in the NOD mouse model using a blocking monoclonal antibody targeting the type 1 diabetes risk-associated T cell co-stimulatory receptor, CD226. Female NOD mice were treated with anti-CD226 at 7-8 weeks of age and then monitored for diabetes incidence and therapeutic mechanism of action. Compared with isotype-treated controls, anti-CD226-treated NOD mice showed reduced insulitis severity (0.84-fold, p=0.0002) at 12 weeks and decreased disease incidence (HR 0.41, p=0.015) at 30 weeks. Flow cytometric analysis performed 5 weeks post treatment demonstrated reduced proliferation of conventional CD4+ T cells (0.87-fold, p=0.030) and CD8+ (0.78-fold, p=0.0018) effector memory T cells in spleens of anti-CD226-treated mice. Phenotyping of pancreatic Tregs revealed increased CD25 expression (2.05-fold, p=0.0073) and signal transducer and activator of transcription 5 (STAT5) phosphorylation (1.39-fold, p=0.0007) following anti-CD226, with splenic Tregs displaying augmented suppression of CD4+ responder T cells (Tresps) (1.49-fold, p=0.0008, 1:2 Treg:Tresp) in vitro. Anti-CD226-treated mice exhibited reduced frequencies of islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)-reactive CD8+ T cells in the pancreas, using both ex vivo tetramer staining (0.50-fold, p=0.0317) and single-cell T cell receptor sequencing (0.61-fold, p=0.022) approaches. 51Cr-release assays demonstrated reduced cell-mediated lysis of beta cells (0.61-fold, p<0.0001, 1:1 effector:target) by anti-CD226-treated autoreactive cytotoxic T lymphocytes. CD226 blockade reduces T cell cytotoxicity and improves Treg function, representing a targeted and rational approach for restoring immune regulation in type 1 diabetes.

Precision Control of Cell Type-Specific Behavior via RNA Sensing and Editing.

In the realms of bioengineering and biopharmaceuticals, there exists a critical demand for advanced genetic tools that can interact with specific cell signaling pathways to accurately identify and target various cell types. This research introduces the innovative CRISPR-ADAReader system, which enables precise manipulation of cell activity through sensing target RNA. Featuring both positive and negative feedback loops, the system allows for tailored regulation across different cell types in response to various internal signals, showcasing exceptional programmability, specificity, and sensitivity. By choosing distinct RNAs as activation signals, the CRISPR-ADAReader efficiently monitors and alters targeted cell behaviors. In a case study focusing on retinoblastoma treatment, the system distinctively initiates positive feedback and self-silencing actions by detecting MCYN and Rb transcripts, thus safeguarding normal retinal pigment epithelial cells while promoting apoptosis in cancer cells. Moreover, the CRISPR-ADAReader demonstrates significant anti-tumor effectiveness in live models, markedly reducing retinoblastoma cell proliferation through the activation of several cancer-suppression pathways, outperforming the capabilities of the ADAR-sensor. Notably, the system also shows an excellent in vivo safety profile. In conclusion, the CRISPR-ADAReader system represents a groundbreaking method for the detection and editing of RNA, offering a potent instrument for the customized and precise governance of cell behavior.

The impact of BTK knockdown on lung adenocarcinoma growth and immune response.

This study explores the molecular role of the BTK gene in lung adenocarcinoma (LUAD) progression and patient prognosis. Using a radiomics model based on BTK expression and PET-CT data analyzed through DeeplabV3, alongside transcriptomic and clinical data from TCGA, we established a strong predictive relationship between BTK levels and LUAD outcomes. Our findings demonstrate that low BTK expression is linked to poorer prognoses. Experimental models, including cell lines and invivo mouse studies, revealed that BTK deficiency leads to increased LUAD cell proliferation, invasion, and metastasis. Furthermore, invivo models indicated that BTK knockdown results in enhanced tumor growth and diminished CD8+ T cell activity. These results suggest that BTK plays a crucial role in modulating LUAD progression and the tumor immune environment, highlighting its potential as a therapeutic target.

Epstein-Barr virus infection upregulates extracellular OLFM4 to activate YAP signaling during gastric cancer progression

Extracellular vesicles (EVs) are known to mediate cell communications and shape tumor microenvironment. Compared to the well-studied small EVs, the function of large microvesicles (MVs) during tumorigenesis is poorly understood. Here we show the proteome of MVs in Epstein-Barr virus (EBV)-associated gastric cancer (EBVaGC), and identify olfactomedin 4 (OLFM4) is induced by EBV infection and secreted via MVs to promote tumor progression through Hippo signaling. Specifically, OLFM4 is a target gene of the cGAS-STING pathway, and EBV infection activates cGAS-STING pathway and increases OLFM4 expression. Moreover, MV-carried OLFM4 binds with the extracellular cadherin domain of FAT1, thereby impairing its intracellular interaction with MST1 and leading to YAP activation in recipient cells. Together, our study not only reveals a regulatory mechanism though which viral infection is coupled via MVs with intercellular control of the Hippo signaling, but also highlights the OLFM4-Hippo axis as a therapeutic target for EBV-associated cancers.

Promoting stem cells activity using Chinese medicine herbs in treatment of neurological disorder: A review on novel therapeutic approaches

Stem cell therapy represents a burgeoning and swiftly advancing modality for the treatment of a diverse array of neurological disorders. However, despite continued clinical trials, the underlying mechanisms of action often remain elusive. Traditional Chinese medicine (TCM), with its holistic approach, provides a valuable resource for the identification and evaluation of potential neuroprotective agents. Research has shown that TCM, including herbs, herbal extracts, and specific Chinese herbal constituents, can modulate the proliferation and differentiation of neural stem cells (NSCs) to some extent. This review examines the potential of TCM as a treatment for neurodegenerative diseases. Given the limitations of current therapies due to a lack of understanding of disease pathogenesis, a holistic approach to TCM offers a promising alternative. This paper also summarizes the role of stem cells in the management of neurological disorders and evaluates prior studies concerning stem cell transplantation. In addition, it explores the capacity of TCM to influence the proliferation and differentiation of NSCs. The ultimate aim of this review is to enhance our understanding of how TCM can be utilized to influence stem cell behavior and potentially treat neurodegenerative diseases.