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Cell Dec 2023Cancer cells are regulated by oncogenic mutations and microenvironmental signals, yet these processes are often studied separately. To functionally map how...
Cancer cells are regulated by oncogenic mutations and microenvironmental signals, yet these processes are often studied separately. To functionally map how cell-intrinsic and cell-extrinsic cues co-regulate cell fate, we performed a systematic single-cell analysis of 1,107 colonic organoid cultures regulated by (1) colorectal cancer (CRC) oncogenic mutations, (2) microenvironmental fibroblasts and macrophages, (3) stromal ligands, and (4) signaling inhibitors. Multiplexed single-cell analysis revealed a stepwise epithelial differentiation phenoscape dictated by combinations of oncogenes and stromal ligands, spanning from fibroblast-induced Clusterin (CLU) revival colonic stem cells (revCSCs) to oncogene-driven LRIG1 hyper-proliferative CSCs (proCSCs). The transition from revCSCs to proCSCs is regulated by decreasing WNT3A and TGF-β-driven YAP signaling and increasing KRAS or stromal EGF/Epiregulin-activated MAPK/PI3K flux. We find that APC loss and KRAS collaboratively limit access to revCSCs and disrupt stromal-epithelial communication-trapping epithelia in the proCSC fate. These results reveal that oncogenic mutations dominate homeostatic differentiation by obstructing cell-extrinsic regulation of cell-fate plasticity.
Topics: Cell Differentiation; Oncogenes; Proto-Oncogene Proteins p21(ras); Signal Transduction; Stem Cells; Humans; Animals; Mice; Cell Lineage
PubMed: 38065080
DOI: 10.1016/j.cell.2023.11.004 -
Cancer Cell Jan 2020Despite substantial clinical benefit of targeted and immune checkpoint blockade-based therapies in melanoma, resistance inevitably develops. We show cytoskeletal...
Despite substantial clinical benefit of targeted and immune checkpoint blockade-based therapies in melanoma, resistance inevitably develops. We show cytoskeletal remodeling and changes in expression and activity of ROCK-myosin II pathway during acquisition of resistance to MAPK inhibitors. MAPK regulates myosin II activity, but after initial therapy response, drug-resistant clones restore myosin II activity to increase survival. High ROCK-myosin II activity correlates with aggressiveness, identifying targeted therapy- and immunotherapy-resistant melanomas. Survival of resistant cells is myosin II dependent, regardless of the therapy. ROCK-myosin II ablation specifically kills resistant cells via intrinsic lethal reactive oxygen species and unresolved DNA damage and limits extrinsic myeloid and lymphoid immunosuppression. Efficacy of targeted therapies and immunotherapies can be improved by combination with ROCK inhibitors.
Topics: Animals; B7-H1 Antigen; Cell Cycle; Cell Line, Tumor; Cytoskeleton; DNA Damage; Drug Resistance, Neoplasm; Female; Humans; Immunotherapy; MAP Kinase Signaling System; Male; Melanoma; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Mice, Nude; Mice, SCID; Myosin Type II; Oxidative Stress; Protein Kinase Inhibitors; Proto-Oncogene Proteins B-raf; Reactive Oxygen Species; T-Lymphocytes, Regulatory; Treatment Outcome; Tumor Microenvironment; rho-Associated Kinases
PubMed: 31935375
DOI: 10.1016/j.ccell.2019.12.003 -
Cell Dec 2023Patient-derived organoids (PDOs) can model personalized therapy responses; however, current screening technologies cannot reveal drug response mechanisms or how tumor...
Patient-derived organoids (PDOs) can model personalized therapy responses; however, current screening technologies cannot reveal drug response mechanisms or how tumor microenvironment cells alter therapeutic performance. To address this, we developed a highly multiplexed mass cytometry platform to measure post-translational modification (PTM) signaling, DNA damage, cell-cycle activity, and apoptosis in >2,500 colorectal cancer (CRC) PDOs and cancer-associated fibroblasts (CAFs) in response to clinical therapies at single-cell resolution. To compare patient- and microenvironment-specific drug responses in thousands of single-cell datasets, we developed "Trellis"-a highly scalable, tree-based treatment effect analysis method. Trellis single-cell screening revealed that on-target cell-cycle blockage and DNA-damage drug effects are common, even in chemorefractory PDOs. However, drug-induced apoptosis is rarer, patient-specific, and aligns with cancer cell PTM signaling. We find that CAFs can regulate PDO plasticity-shifting proliferative colonic stem cells (proCSCs) to slow-cycling revival colonic stem cells (revCSCs) to protect cancer cells from chemotherapy.
Topics: Humans; Apoptosis; Cancer-Associated Fibroblasts; Organoids; Signal Transduction; Single-Cell Analysis; Drug Evaluation, Preclinical; Algorithms; Stem Cells
PubMed: 38065081
DOI: 10.1016/j.cell.2023.11.005 -
Dermatitis : Contact, Atopic,... Oct 2021Tape-strips are a minimally invasive approach to characterize skin biomarkers in atopic dermatitis (AD). However, they have not yet been used for tracking gene...
BACKGROUND
Tape-strips are a minimally invasive approach to characterize skin biomarkers in atopic dermatitis (AD). However, they have not yet been used for tracking gene expression changes with systemic treatment.
OBJECTIVE
The aim of the study was to evaluate gene expression changes and therapeutic response biomarkers in AD patients before and after dupilumab (interleukin 4Rα antibody) treatment using tape-strips to obtain epidermal tissue for analysis.
METHODS
Lesional and nonlesional tape-stripped skin was sampled from 18 AD patients before and after dupilumab treatment and from 17 healthy subjects and analyzed by RNA-seq.
RESULTS
At baseline, we detected 6745 and 4859 differentially expressed genes between lesional and nonlesional skin versus normal, respectively, whereas 841 and 977 genes were differentially expressed after treatment, respectively (fold change >1.5 and false discovery rate <0.05). Tape-strips captured significant modulation with dupilumab in key AD immune (eg, C-C motif chemokine ligand 13 [CCL13], CCL17, CCL18) and barrier (eg, periplakin, FA2H) biomarkers. Changes in biomarkers (CCL20, interleukin 34, FABP7) were also significantly correlated with clinical disease improvements (Eczema Area and Severity Index; R > 0.5 or R < -0.4, P < 0.05).
CONCLUSIONS
This real-life study represents the first comprehensive RNA-seq molecular profiling of tape-strips from moderate to severe AD patients after dupilumab therapy. Analysis of tape strip specimens detected significant gene expression changes in key AD biomarkers with dupilumab treatment, suggesting that this approach may be useful to monitor therapeutic responses in inflammatory skin diseases.
Topics: Adult; Antibodies, Monoclonal, Humanized; Case-Control Studies; Dermatitis, Atopic; Female; Gene Expression Profiling; Humans; Male; Middle Aged; Sequence Analysis, RNA; Skin
PubMed: 34405829
DOI: 10.1097/DER.0000000000000764 -
Obesity (Silver Spring, Md.) Jun 2022Three-dimensional (3D) imaging systems are increasingly being used in health care settings for quantifying body size and shape. The potential exists to provide similar...
OBJECTIVE
Three-dimensional (3D) imaging systems are increasingly being used in health care settings for quantifying body size and shape. The potential exists to provide similar phenotyping capabilities outside of professional settings using smartphone applications (apps). The current study aim was to compare waist, hip, upper arm, and midthigh circumference measurements acquired by a free downloadable app (MeThreeSixty; Size Stream, Cary, North Carolina) and a conventional 20-camera 3D system (SS20; Size Stream) with those measured with a flexible tape at the same anatomic sites.
METHODS
Fifty-nine adults were scanned with the app and SS20; the same software was used to generate circumference estimates from device-acquired object files that were then compared with reference tape measurements.
RESULTS
The app and SS20 had similar coefficients of variation that were minimally larger than those by the tape (e.g., waist, 0.93%, 0.87%, and 0.06%). Correlations of the app and of SS20 with tape circumferences were all strong (p < 0.001) and similar in magnitude (R s: 0.72-0.93 and 0.78-0.95, respectively); minimally significant (p < 0.05 to p < 0.01) bias was present between both imaging approaches and some tape measurements.
CONCLUSION
These proof-of-concept observations combined with ubiquitous smartphone availability create the possibility of phenotyping adult body size and shape, with important clinical and research implications, on a global scale.
Topics: Anthropometry; Body Size; Mobile Applications; Smartphone
PubMed: 35491718
DOI: 10.1002/oby.23434 -
Nano Letters Oct 2021Photothermal bimorph actuators have attracted considerable attention in intelligent devices because of their cordless control and lightweight and easy preparation....
Photothermal bimorph actuators have attracted considerable attention in intelligent devices because of their cordless control and lightweight and easy preparation. However, current photothermal bimorph actuators are mostly based on films or papers driven by near-infrared sources, which are deficient in flexibility and adaptability, restricting their potential in wearable applications. Herein, a bimorph textile actuator that can be scalably fabricated with a traditional textile route and autonomously triggered by sunlight is reported. The active layer and passive layer of the bimorph are constructed by polypropylene tape and a MXene-modified polyamide filament. Because of the opposite thermal expansion and MXene-enhanced photothermal efficiency (>260%) of the bimorph, the textile actuator presents effective deformation (1.38 cm) under low sunlight power (100 mW/cm). This work provides a new pathway for wearable sunlight-triggered actuators and finds attractive applications for smart textiles.
Topics: Nylons; Polypropylenes; Sunlight; Textiles; Wearable Electronic Devices
PubMed: 34570519
DOI: 10.1021/acs.nanolett.1c02578 -
An Investigation Into Different Measurement Techniques to Assess Equine Proximal Hoof Circumference.Journal of Equine Veterinary Science Aug 2022Equine hoof conformation is integral to equine performance and soundness. Consequently, it is a major area of interest within the field of equine health. Researchers...
Equine hoof conformation is integral to equine performance and soundness. Consequently, it is a major area of interest within the field of equine health. Researchers have measured several hoof shape parameters to study the hoof conformation. Proximal hoof circumference (PHC) is a primary hoof shape parameter, and its assessment may help to recognize the early stages of the development of changes in hoof morphology or poor hoof shape. Previous studies have mainly used a measuring tape to measure PHC. However, some doubts still exist regarding the reliability, repeatability and accuracy of measuring tape in this context. The current study conducted a technical comparison between the measuring tape and two alternative methods of 3D scanning and photogrammetry to measure PHC. Five equine limbs were collected from five adult horses, and the PHC of the limbs was measured using these three methods. The 3D scanner method was considered to be the highest accuracy and the reference for method comparisons. Pairwise correlations between the 3D scanner and the other two methods were conducted using a linear mixed model. The measuring tape and photogrammetry tended to overestimate the mean PHC compared to the 3D scanner by 0.96 mm (P > .05) and 2.2 mm (P < .05), respectively. In addition, an excellent interrater and intrarater correlation coefficient index was reported for the reliability of the tape measurements. The variation of the tape measurements was ±2 mm, which justified the use of measuring tape for PHC measurements in various clinical and horse management applications.
Topics: Animals; Extremities; Hoof and Claw; Horses; Photogrammetry; Reproducibility of Results
PubMed: 35661772
DOI: 10.1016/j.jevs.2022.104028 -
Sensors (Basel, Switzerland) Oct 2022The role of humidity sensors in different industries and field applications, such as agriculture, food monitoring, biomedical equipment, heating, and ventilation, is...
The role of humidity sensors in different industries and field applications, such as agriculture, food monitoring, biomedical equipment, heating, and ventilation, is well known. However, most commercially available humidity sensors are based on polymers or electronic materials that are not degradable and thus contribute to electronic waste. Here, we report a low-cost, flexible, easy-to-fabricate, and eco-friendly parallel-plate capacitive humidity sensor for field applications. The sensor is fabricated from copper tape and tissue paper, where copper tape is used to create the plates of the capacitor, and tissue paper is used as a dielectric sensing layer. Along with the low cost, the high sensitivity, better response and recovery times, stability, and repeatability make this sensor unique. The sensor was tested for relative humidity (RH), ranging from 40% to 99%, and the capacitance varied linearly with RH from 240 pF to 720 pF, as measured by an Arduino. The response time of the sensor is ~1.5 s, and the recovery time is ~2.2 s. The experiment was performed 4-5 times on the same sensor, and repeatable results were achieved with an accuracy of ±0.1%. Furthermore, the sensor exhibits a stable response when tested at different temperatures. Due to the above advantages, the presented sensor can find ready applications in different areas.
Topics: Humidity; Copper; Electric Capacitance; Electronics; Polymers
PubMed: 36298240
DOI: 10.3390/s22207885 -
Biomaterials Science Dec 2021We describe the synthesis of poly(glycidyl acetate--glycidyl butyrate carbonate)s the terpolymerization of glycidyl acetate (GA), glycidyl butyrate (GB), and CO by a...
We describe the synthesis of poly(glycidyl acetate--glycidyl butyrate carbonate)s the terpolymerization of glycidyl acetate (GA), glycidyl butyrate (GB), and CO by a cobalt salen complex in high atom economy. These new non-cytotoxic polycarbonates are pressure-sensitive adhesives, and peel testing shows the adhesive strength ranges from Scotch-Tape® to hot-melt glues based on glycidyl butyrate content. The tunable adherence, benign degradation products, and facile application and removal suggest their utility as temporary adhesives, such as those used in biomedical applications or medical devices. One polymer, (GA--GB)-87, exhibits the proper adhesive strength to sufficiently adhere a collagen buttress to the jaws of a steel surgical stapler and easily release the buttress after firing to successfully cut, close, and implant the buttress into lung tissue in an sheep model.
Topics: Adhesives; Animals; Cobalt; Glycerol; Materials Testing; Polymers; Sheep; Tissue Adhesives
PubMed: 34787119
DOI: 10.1039/d1bm00995h -
Current Protocols Mar 2021Staphylococcus aureus is a Gram-positive bacterium that colonizes almost every organ in humans and mice and is a leading cause of diseases worldwide. S. aureus...
Staphylococcus aureus is a Gram-positive bacterium that colonizes almost every organ in humans and mice and is a leading cause of diseases worldwide. S. aureus infections can be challenging to treat due to widespread antibiotic resistance and their ability to cause tissue damage. The primary modes of transmission of S. aureus are via direct contact with a colonized or infected individual or invasive spread from a colonization niche in the same individual. S. aureus can cause a myriad of diseases, including skin and soft tissue infections (SSTIs), osteomyelitis, pneumonia, endocarditis, and sepsis. S. aureus infection is characterized by the formation of purulent lesions known as abscesses, which are rich in live and dead neutrophils, macrophages, and surrounded by a capsule containing fibrin and collagen. Different strains of S. aureus produce varying amounts of toxins that evade and/or elicit immune responses. Therefore, animal models of S. aureus infection provide a unique opportunity to understand the dynamics of organ-specific immune responses and modifications in the pathogen that could favor the establishment of the pathogen. With advances in in vivo imaging of fluorescent transgenic mice, combined with fluorescent/bioluminescent bacteria, we can use mouse models to better understand the immune response to these types of infections. By understanding the host and bacterial dynamics within various organ systems, we can develop therapeutics to eliminate these pathogens. This module describes in vivo mouse models of both local and systemic S. aureus infection. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Murine model of Staphylococcus aureus subcutaneous infection Alternate Protocol: Murine tape stripping skin infection model Basic Protocol 2: Sample collection to determine skin structure, production of inflammatory mediators, and bacterial load Basic Protocol 3: Murine model of post-traumatic Staphylococcus aureus osteomyelitis Basic Protocol 4: Intravenous infection of the retro-orbital sinus Support Protocol: Preparation of the bacterial inoculum.
Topics: Animals; Disease Models, Animal; Mice; Mice, Transgenic; Soft Tissue Infections; Staphylococcal Infections; Staphylococcus aureus
PubMed: 33656290
DOI: 10.1002/cpz1.52