Cell Behavior Research Articles

Synergic effects of core-shell nanospheres and magnetic field for sciatic nerve regeneration in decellularized artery conduits with Schwann cells

Numerous conduits have been developed to improve peripheral nerve regeneration. However, challenges remain, including remote control of conduit function, and programmed cell behaviors like orientation. We synthesized Fe3O4-MnO2@Zirconium-based Metal-organic frameworks@Retinoic acid (FMZMR) core-shell and assessed their impact on Schwann cell function and behavior within conduits made from decellularized human umbilical arteries (DHUCA) under magnetic field (MF). FMZMR core-shell, featuring a spherical porous structure and catalytic properties, effectively scavenges radicals and facilitates controlled drug release under MF. The histology of the DHUCA indicates effective decellularization with adequate tensile strength and Young’s modulus for sciatic nerve regeneration. In-vitro results demonstrate that FMZMR core-shell is biocompatible and promotes Schwann cell proliferation through remotely controlled drug release. Furthermore, its synergy with MF enhances cell orientation and increases neurite length by ~ 1.93-fold. Functional and histological evaluations indicate that the FMZMR core-shell combined with MF promotes nerve regeneration, decreases muscle atrophy, and enhances new neuron growth and myelin formation, without negatively affecting vital tissues. This study suggests that the synergistic effect of FMZMR core-shell with MF can alleviate some of the treatment challenges.Graphical

Methyltransferase like 13 promotes malignant behaviors of bladder cancer cells through targeting PI3K/ATK signaling pathway

Abstract Bladder cancer (BC) is the tenth most common tumor worldwide, characterized by high incidence rates and mortality. This study aimed to explore the role of Methyltransferase like 13 (METTL13) in BC cells. J82 and T24 cells were cultured for in vitro experiments. Cell viability, migration, and invasion were assessed using CCK-8 and transwell assays. Senescence-associated beta-galactosidase (SA-β-gal) levels were detected using a β-galactosidase staining kit. METTL13 and cell cycle-related protein levels were quantified using RT-qPCR and Western blotting. The results showed that METTL13 was upregulated in BC cells. Silencing METTL13 decreased cell viability, migration, and invasion in BC cells, whereas METTL13 overexpression increased these parameters. Additionally, METTL13 knockdown inhibited the phosphorylation levels of PI3K, AKT, and mTOR. Inhibition of the PI3K/AKT pathway reversed the effects of METTL13 on cell viability, migration, invasion, and cell cycle-related proteins in BC cells. In vivo experiments showed that METTL13 knockdown inhibited tumor growth and development. In conclusion, this study demonstrated that METTL13 promoted the malignant behaviors of BC cells through activation of the PI3K/AKT signaling pathway. METTL13 may be a promising therapeutic target for BC in the future.

GATA1 activates HSD17B6 to improve efficiency of cisplatin in lung adenocarcinoma via DNA damage

BackgroundLung adenocarcinoma (LUAD) is the most common histological type of non-small cell lung cancer (NSCLC). Platinum-based chemotherapy, such as cisplatin chemotherapy, is the cornerstone of treatment for LUAD patients. Nevertheless, cisplatin resistance remains the key obstacle to LUAD treatment, for its mechanism has not been fully elucidated.MethodsHSD17B6 mRNA expression data were accessed from TCGA-LUAD database and differential expression analysis was performed. Enrichment analysis of HSD17B6 was conducted by GSEA, and its upstream transcription factors were predicted by hTFtarget. mRNA and protein expression levels of HSD17B6 and GATA1 were assayed by qRT-PCR and WB, and the binding relationship between them was verified by chromatin immunoprecipitation assay and dual luciferase reporter assay. Cell viability and IC50 value of cisplatin-treated cells were measured by cell counting kit-8 assay. Cell cycle was assayed by flow cytometry. DNA damage level and DNA damage marker γ-H2AX expression were assayed by comet assay and western blot, respectively.ResultsHSD17B6 was lowly expressed in LUAD tissues and cells and mainly enriched in homologous recombination and mismatch repair pathways. As cell function experiments revealed, overexpression of HSD17B suppressed malignant phenotypes and cisplatin resistance in LUAD cells through DNA damage. Bioinformatics analysis revealed that GATA1 is the upstream regulator of HSD17B6, which was markedly reduced in LUAD tissues and cells. ChIP and dual luciferase reporter assays ascertained the binding of GATA1 to HSD17B6. Knockdown of GATA1 attenuated the effect of overexpression of HSD17B6 on LUAD cell behaviors and cisplatin resistance.ConclusionTranscription factor GATA1 could activate HSD17B6 to inhibit cisplatin resistance in LUAD through DNA damage, suggesting that GATA1/HSD17B6 axis may be a potential therapeutic target for chemotherapy resistance in LUAD patients.

High-Cycle Fatigue Behaviour and Structural Robustness of Glass Fibre-Reinforced Polymer Tiled Web-Core Sandwich Panel Unit Cells in Load-Bearing Structures

This paper explores the fatigue behaviour and robustness of tiled web-core sandwich panels used in glass fibre-reinforced polymer bridges, which are increasingly favoured for their lightweight and corrosion-resistant properties. Fatigue tests are conducted on unit cell specimens with manually induced crack initiation, simulating accidental damage scenarios in glass fibre-reinforced polymer bridge components. The objective is to assess the integrity of individual unit cells when subjected to a localized force at the top flange after damage initiation. The fatigue tests reveal three phases in the behaviour of a tiled unit cell. Initially, there is a substantial rapid stiffness degradation with crazing crack appearance within the cross-section. Subsequently, a plateau phase occurs, with limited stiffness degradation and stable crazing cracks, the duration of which depends on the applied fatigue load. Lastly, rapid stiffness degradation with substantial crack growth leads to ultimate failure within roughly a thousand cycles. Further analysis using digital image correlation reveals strain concentrations at the location of crazing cracks and crack propagation occurring interlaminarly but not through the plies of the top and bottom flanges, ensuring a robust design. This research enhances the understanding of the tiled sandwich panels, offering prospects for resilient load-bearing structures in glass fibre-reinforced polymer bridges and structural engineering applications.

Myosin Light Chains in the Progression of Cancer

The myosin light chains (MLCs) of non-muscle myosin II are known to regulate cellular architecture and generate cellular forces; they also have an increasingly emerging role in the progression of cancer. The phosphorylation state of the myosin light chains controls the activity of myosins that are implicated in invasion and proliferation. In cancers, when proliferation is greatly increased, cytokinesis relies on phosphorylated light chains to activate the contractile forces used to separate the cells. Likewise, during metastasis, kinase pathways culminate in aligning MLC structures for enhanced cell motility through stress fiber contraction and the accumulation of myosin filaments at the leading edge. This review summarizes the myosin light chain family members known to promote cancer progression and evidence of how their altered activities change the behavior of cells involving the mechanical-based processes of proliferation and cell movements during metastasis. In addition, myosin light chains impact the immune response to cancers and currently serve as biomarkers in staging this disease; a brief summary of these topics is provided at the end of the review.

Fibroblasts modulate epithelial cell behavior within the proliferative niche and differentiated cell zone within a human colonic crypt model

Colonic epithelium is situated above a layer of fibroblasts that provide supportive factors for stem cells at the crypt base and promote differentiation of cells in the upper crypt and luminal surface. To study the fibroblast-epithelial cell interactions, an in vitro crypt model was formed on a shaped collagen scaffold with primary epithelial cells growing above a layer of primary colonic fibroblasts. The crypts possessed a basal stem cell niche populated with proliferative cells and a differentiated, nondividing cell zone at the luminal crypt end. The presence of fibroblasts enhanced cell differentiation and accelerated the rate at which a high resistance epithelial cell layer formed relative to cultures without fibroblasts. The fibroblasts modulated cell proliferation within crypts increasing the number of crypts populated with proliferative cells but decreasing the total number of proliferative cells in each crypt. Bulk-RNA sequencing revealed 41 genes that were significantly upregulated and 190 genes that were significantly downregulated in cocultured epithelium relative to epithelium cultured without fibroblasts. This epithelium-fibroblast crypt model suggests bidirectional communication between the two cell types and has the potential to serve as a model to investigate fibroblast-epithelial cell interactions in health and disease.

Gate-Tunable High-Responsivity Photodiode Based on 2D Ambipolar Semiconductor

Abstract Electrically tunable homojunctions based on ambipolar two-dimensional materials have attracted widespread attention in the field of intelligent vision. These devices exhibit inherent switchable positive and negative photovoltaic properties, that effectively mimic the behavior of human retinal cells. However, the photovoltaic responsivity of most electrically tunable homojunctions remain significantly low due to the weak light absorption, making it challenging to meet the application requirements for high-sensitivity target detection in the field of intelligent vision. Here, we propose a gate-tunable photodiode based on two-dimensional ambipolar WSe2 with an asymmetric gate electrode, achieving high photovoltaic responsivity. By adjusting the gate voltage and keeping bias voltage zero, we can dynamically realize reconfigurable n--p and n--n homojunction states, as well as gate-tunable photovoltaic response characteristics that range from positive to negative. The maximum photovoltaic responsivity of the electrically tunable WSe2 homojunction is approximately 0.4 A/W, which is significantly larger than the previously reported value ~0.1 A/W in homojunction devices. In addition, the responsivity can be further enhanced to approximately 1.0 A/W when the n--p photodiode operates in reverse bias mode, enabling high-sensitivity detection of targets. Our work paves the way for developing gate-tunable photodiodes with high photovoltaic responsivity and advancing high-performance intelligent vision technology.

Peptide Designs for Cell-Interfacing Assemblies.

Constructing synthetic materials to interact with cells offers significant promise for understanding natural cellular processes and manipulating cell behaviors beyond nature's capabilities. Peptide assemblies are particular promising in this regard, as they have demonstrated efficacy in promoting cell differentiation, repair, and regeneration as supportive scaffolds. However, distinct gaps persist between natural and synthetic assembly systems in various aspects. This Concept review explores representative studies focusing on developing chemical strategies to design peptide assemblies with precise control over displayed molecules, adjustable molecular dynamics to modulate cell behaviors, and responsiveness to biological stimuli. This paper would inspire more fundamental studies on molecular assemblies and foster inter-disciplinary interests in material applications, advancing the interplay between natural cells and synthetic materials.

Human–machine interfaces and the NEU-CHiP Project: an interview with Dr Eric Hill

After spending 8 years developing his laboratory at Aston University, Dr Eric Hill is now a Reader in Cellular and Molecular Neurobiology at Loughborough University. His main research interests include the development of tissue engineering strategies to model stem cell behaviour in the development of neuronal networks and also during neurodegeneration.

Integrating Physical and Biochemical Cues for Muscle Engineering: Scaffolds and Graft Durability

Muscle stem cells (MuSCs) are essential for skeletal muscle regeneration, influenced by a complex interplay of mechanical, biochemical, and molecular cues. Properties of the extracellular matrix (ECM) such as stiffness and alignment guide stem cell fate through mechanosensitive pathways, where forces like shear stress translate into biochemical signals, affecting cell behavior. Aging introduces senescence which disrupts the MuSC niche, leading to reduced regenerative capacity via epigenetic alterations and metabolic shifts. Transplantation further challenges MuSC viability, often resulting in fibrosis driven by dysregulated fibro-adipogenic progenitors (FAPs). Addressing these issues, scaffold designs integrated with pharmacotherapy emulate ECM environments, providing cues that enhance graft functionality and endurance. These scaffolds facilitate the synergy between mechanotransduction and intracellular signaling, optimizing MuSC proliferation and differentiation. Innovations utilizing human pluripotent stem cell-derived myogenic progenitors and exosome-mediated delivery exploit bioactive properties for targeted repair. Additionally, 3D-printed and electrospun scaffolds with adjustable biomechanical traits tackle scalability in treating volumetric muscle loss. Advanced techniques like single-cell RNA sequencing and high-resolution imaging unravel muscle repair mechanisms, offering precise mapping of cellular interactions. Collectively, this interdisciplinary approach fortifies tissue graft durability and MuSC maintenance, propelling therapeutic strategies for muscle injuries and degenerative diseases.

Framework Nucleic Acids: Innovative Tools for Cellular Sensing and Therapeutics.

As emerging biomaterials, framework nucleic acids (FNAs) have recently demonstrated great potential in the biomedical field due to their high programmability, biocompatibility, unique structural diversity, and precise molecular design capabilities. This review focuses on the applications of FNAs in cellular sensing and disease treatment. First, we systematically introduce the applications of FNAs in cellular sensing, including their precise recognition and response to the extracellular tumor microenvironment, cell membrane proteins, and intracellular biomarkers. Subsequently, we review the potential of FNAs in disease treatment, covering their applications and development in drug delivery, regulation of cell behavior, and immunomodulation. We also discuss the limitations and potential role of FNAs in personalized medicine, precision diagnostics, and advanced therapies. The broad application of FNAs is expected to drive significant breakthroughs in future biomedical technological innovations and clinical translation.

Exploring Microgravity’s Effect on Seed Germination and Growth of Plants

The study of the effects of microgravity on plant cells has gained significant attention, especially in the context of space experiments, which necessitates a comprehensive understanding of plant growth and development in altered gravitational conditions. Understanding the impact of microgravity on seed germination, seedling establishment, and overall plant development is essential for future space agriculture and the development of sustainable food production systems. In microgravity, plants encounter unique physical and physiological challenges, making it essential to explore the physiological and biochemical responses of plant cells to this environment. Various factors contribute to the alterations observed in plant behavior under microgravity, including changes in water and nutrient distribution, disruptions in hormone signaling, and modifications in the expression of secondary metabolites. Physiological and biochemical in plants due to microgravity caused by the influence on the phytochrome system, gas exchange photosynthesis, and the production of secondary metabolites. Microgravity affects mechanocellular responses of plants which elucidates how the absence of gravity can disrupt cytoplasmic streaming, impact cell wall rigidity, and cause damage to cell membranes. These alterations in plant cell behavior under microgravity conditions can have profound implications for plant growth and development. Space farming faces challenges and constraints of cultivating plants in space, such as managing water distribution, temperature fluctuations, and radiation exposure, are addressed. Innovative methods for planting crops in space, including plant pillows, greenhouse cultivation, and soil-less systems, are discussed. In the future, research efforts should focus on selecting or genetically modifying plant varieties with enhanced tolerance to the diverse stress encountered in space environments. Thus, it is concluded that space farming underscores the importance of comprehensive studies on plant responses to microgravity, offering valuable insights for both space exploration and terrestrial agriculture.

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.

In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel.

Chronic lymphocytic leukemia (CLL) is a common adult leukemia characterized by the accumulation of neoplastic mature B cells in blood, bone marrow, lymph nodes, and spleen. The disease biology remains unresolved in many aspects, including the processes underlying the disease progression and relapses. However, studying CLL in vitro poses a considerable challenge due to its complexity and dependency on the microenvironment. Several approaches are utilized to overcome this issue, such as co-culture of CLL cells with other cell types, supplementing culture media with growth factors, or setting up a three-dimensional (3D) culture. Previous studies have shown that 3D cultures, compared to conventional ones, can lead to enhanced cell survival and altered gene expression. 3D cultures can also give valuable information while testing treatment response in vitro since they mimic the cell spatial organization more accurately than conventional culture. In our study, we investigated the behavior of CLL cells in two types of material: (i) solid porous collagen scaffolds and (ii) gel composed of carboxymethyl cellulose and polyethylene glycol (CMC-PEG). We studied CLL cells' distribution, morphology, and viability in these materials by a transmitted-light and confocal microscopy. We also measured the metabolic activity of cultured cells. Additionally, the expression levels of MYC, VCAM1, MCL1, CXCR4, and CCL4 genes in CLL cells were studied by qPCR to observe whether our novel culture approaches lead to increased adhesion, lower apoptotic rates, or activation of cell signaling in relation to the enhanced contact with co-cultured cells. Both materials were biocompatible, translucent, and permeable, as assessed by metabolic assays, cell staining, and microscopy. While collagen scaffolds featured easy manipulation, washability, transferability, and biodegradability, CMC-PEG was advantageous for its easy preparation process and low variability in the number of accommodated cells. Both materials promoted cell-to-cell and cell-to-matrix interactions due to the scaffold structure and generation of cell aggregates. The metabolic activity of CLL cells cultured in CMC-PEG gel was similar to or higher than in conventional culture. Compared to the conventional culture, there was (i) a lower expression of VCAM1 in both materials, (ii) a higher expression of CCL4 in collagen scaffolds, and (iii) a lower expression of CXCR4 and MCL1 (transcript variant 2) in collagen scaffolds, while it was higher in a CMC-PEG gel. Hence, culture in the material can suppress the expression of a pro-apoptotic gene (MCL1 in collagen scaffolds) or replicate certain gene expression patterns attributed to CLL cells in lymphoid organs (low CXCR4, high CCL4 in collagen scaffolds) or blood (high CXCR4 in CMC-PEG).

3D printed β-TCP scaffolds loaded with SVVYGLR peptide for promoting revascularization and osteoinduction

It is crucial for the successful transplantation of large segmental bone defects to achieve rapid vascularization within bone scaffolds. However, there are certain limitations including uncontrolled angiogenesis and inadequate vascular function. Therefore, there is an urgent need to develop bone scaffolds with functional vascular networks. In our study, porousβ-tricalcium phosphate (β-TCP) scaffolds with varying pore sizes were prepared by 3D printing technology, loaded with osteopontin derived peptide Ser-Val-Val-Tyr-Gly-Leu-Arg (SVVYGLR) to induce osteoinduction and angiogenesis.In vitro, the proliferation and migration behaviors of human umbilical vein endothelial cell on scaffolds were assessed by Cell Counting Kit-8, confocal laser scanning microscopy and scanning electron microscopy. And the osteogenic ability of bone marrow mesenchymal stem cells was assessed using alkaline phosphatase staining and Alizarin Red S staining. The messenger ribonucleic acid (mRNA) expression levels of cell adhesion molecule (CD31), vascular endothelial growth factor and hypoxia inducible factor-1αin each group were detected by quantitative real-time fluorescence polymerase chain reaction (PCR) analysis.In vivo, cube scaffolds were subcutaneously implanted on the right hips of Sprague-Dawley (SD) rats for 6 weeks. Hematoxylin and Eosin staining, Masson's trichrome staining, and immunohistochemical analysis of osteocalcin and CD31 were performed on slices for every sample with three sections to explore the effect of SVVYGLR-loaded scaffolds on angiogenesis and osteogenic induction for bone reconstruction. The results indicate that 3D printedβ-TCP scaffolds loaded with the SVVYGLR peptide offer superior revascularization and osteoinduction to the scaffolds without the SVVYGLRin situ. Moreover, scaffolds with a pore size of 400 µm demonstrate higher effectiveness compared to those with a 150 µm pore size. The distinct hollow channel scaffolds and the specific SVVYGLR peptide substantially improve cell adhesion, spreading, and proliferation, as well as promote angiogenesis and bone formation. Furthermore, scaffolds with a pore size of 400 µm may exhibit greater efficacy compared to those with a pore size of 150 µm. The results of this study provide an idea for the development of practical applications for tissue-engineered bone scaffolds.

LINC01614 Accelerates CRC Progression via STAT1/LINC01614/miR-4443/PFKFB3-Mediated Aerobic Glycolysis.

Colorectal cancer (CRC) is an aggressive malignancy among malignant tumours, with a high incidence globally. LINC01614, a long non-coding RNA, has been identified as an essential regulator in multiple cancer types. However, its biological functions and underlying molecular mechanisms in CRC remain largely unknown. In this study, we employed RT-qPCR to assess the expression levels of LINC01614 in CRC samples. In vitro, glucose metabolism experiments were conducted to evaluate glucose metabolism in cells. The binding relationship between miR-4443, PFKFB3, and LINC01614 was confirmed through fluorescence reporter gene detection. The subcellular localization of LINC01614 in CRC cells was determined using FISH and subcellular fractionation experiments. Additionally, a mouse subcutaneous tumor model was established for in vivo experiments. Our findings reveal that LINC01614 is upregulated in CRC tissues. Silencing of LINC01614 suppresses the malignant behaviors of CRC cells, including cell proliferation, invasion, migration, and aerobic glycolysis. Furthermore, we discovered that LINC01614 promotes the expression of PFKFB3. Additional experiments demonstrated that LINC01614 binds to miR-4443, leading to the upregulation of PFKFB3 expression. Further experiments confirmed that the LINC01614/miR-4443/PFKFB3 axis promotes CRC cell malignancy by enhancing aerobic glycolysis. Additionally, we found that STAT1 promotes the transcription of LINC01614. These findings uncover a novel regulatory pathway wherein STAT1-induced LINC01614 enhances PFKFB3 expression by sponging miR-4443, thereby accelerating CRC development. This understanding may lead to novel therapeutic strategies for CRC treatment.

Decellularized Cell‐Secreted Extracellular Matrices as Biomaterials for Tissue Engineering

The extracellular matrix (ECM) is the naturally secreted biomaterial scaffold that provides support and regulates key aspects of cell behavior. This dynamic and complex network of structural proteins, proteoglycans, and soluble cues defines the cell microenvironment and is essential for tissue homeostasis. Because tissue engineering approaches aim to recapitulate aspects of the microenvironment to instruct tissue regeneration, ECM‐inspired or ‐derived scaffolds are some of the earliest tissue‐engineered constructs reported. However, conventional single‐protein constructs fail to provide the biochemical and structural complexity of the native ECM. Decellularized ECM is under investigation to improve cell adhesion, cell remodeling, migration, proliferation, and differentiation within tissue‐engineered constructs. However, challenges associated with poor mechanical properties and inherent chemical instability compared to synthetic or other natural polymers require additional considerations. This review describes the bioactive properties of ECM, current strategies to efficiently decellularize cell‐secreted and tissue‐derived ECM, standard fabrication techniques for ECM constructs, and current developments in the field of ECM‐based musculoskeletal platforms.

Modeling Thermal Runaway Mechanisms and Pressure Dynamics in Prismatic Lithium-Ion Batteries

Lithium-ion batteries play a vital role in modern energy storage systems, being widely utilized in devices such as mobile phones, electric vehicles, and stationary energy units. One of the critical challenges with their use is the thermal runaway (TR), typically characterized by a sharp increase in internal pressure. A thorough understanding and accurate prediction of this behavior are crucial for improving the safety and reliability of these batteries. To achieve this, two new combined models were developed: one to simulate the thermal runaway and another to simulate the internal cell pressure. The thermal model tracks a chain of decomposition reactions that eventually lead to TR. At the same time, the pressure model simulates the proportional increase in pressure due to the evaporation of the electrolyte and the gases produced from the decomposition reactions. What sets this work apart is the validation of the pressure model through experimental data, specifically for prismatic lithium-ion cells using NMC chemistries with varying stoichiometries—NMC111 and NMC811. While the majority of the literature focuses on the simulation of temperature and pressure for cylindrical cells, studies addressing these aspects in prismatic cells are much less common. This article addresses this gap by conducting pressure validation experiments, which are hardly documented in the existing studies. Furthermore, the model’s accuracy and flexibility are tested through two experiments, conducted under diverse conditions to ensure robust and adaptive predictions of cell behavior during failure scenarios.

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.

Exploring Acoustic Detection of α-Synuclein Fibrils.

Over the past decades, the incidence of Parkinson's disease (PD) cases has doubled in industrialized countries. While patients over 70years old still represent more than half of the cases, the disease is increasingly affecting younger individuals. Environmental factors have been implicated, such as the effects of certain pesticides or chemicals on neurons, such as rotenone or 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Researchers have also demonstrated the influence of genetic mutations in younger patients. A-synuclein is a protein encoded by the SNCA gene, known to undergo various mutations in hereditary cases of PD. These mutations alter the composition and spatial arrangements of α-synuclein. The proteins, originally of linear shape, aggregate during the progression of PD, forming fibrillary structures that propagate through brain tissues. Among the physical therapies investigated for treating α-synuclein aggregation, ultrasonic waves, capable of altering protein and cell behaviors, have recently been used to disrupt α-synuclein fibrils within tissues in cellular and animal models, with the hope of developing treatments based on ultrasound properties. However, detecting fibrils typically requires invasive and non-biocompatible chemical compounds or cumbersome machinery. In this study, our acoustic experimental setup allowed us to investigate the response of α-synuclein to ultrasound perturbations. By capturing the transmitted wave across proteins over a frequency range 10kHz to 10MHz, no ultrasound signature indicating the presence of proteins was observed.Significance Statement: The results report there is no ultrasound signature of the presence of α-synuclein fibrils, from 10kHz to 10MHz.