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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 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 -
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 -
Computer Methods and Programs in... Jul 2020Conventional anthropometric studies using Kinect depth sensors have concentrated on estimating the distances between two points such as height. This paper deals with a...
BACKGROUND AND OBJECTIVE
Conventional anthropometric studies using Kinect depth sensors have concentrated on estimating the distances between two points such as height. This paper deals with a novel waist measurement method using SVM regression, further widening spectrum of Kinect's potential applications. Waist circumference is a key index for the diagnosis of abdominal obesity, which has been linked to metabolic syndromes and other related diseases. Yet, the existing measuring method, tape measure, requires a trained personnel and is therefore costly and time-consuming.
METHODS
A dataset was constructed by recording both 30 frames of Kinect depth image and careful tape measurement of 19 volunteers by a clinical investigator. This paper proposes a new SVM regressor-based approach for estimating waist circumference. A waist curve vector is extracted from a raw depth image using joint information provided by Kinect SDK. To avoid overfitting, a data augmentation technique is devised. The 30 frontal vectors and 30 backside vectors, each sampled for 1 s per person, are combined to form 900 waist curve vectors and a total of 17,100 samples were collected from 19 individuals. On an individual basis, we performed leave-one-out validation using the SVM regressor with the tape measurement-gold standard of waist circumference measurement-values labeled as ground-truth. On an individual basis, we performed leave-one-out validation using the SVM regressor with the tape measurement-gold standard of waist circumference measurement-values labeled as ground-truth.
RESULTS
The mean error of the SVM regressor was 4.62 cm, which was smaller than that of the geometric estimation method. Potential uses are discussed.
CONCLUSIONS
A possible method for measuring waist circumference using a depth sensor is demonstrated through experimentation. Methods for improving accuracy in the future are presented. Combined with other potential applications of Kinect in healthcare setting, the proposed method will pave the way for patient-centric approach of delivering care without laying burdens on patients.
Topics: Anthropometry; Body Mass Index; Female; Humans; Japan; Male; Obesity; Support Vector Machine; Waist Circumference
PubMed: 32126448
DOI: 10.1016/j.cmpb.2020.105418 -
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 -
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 -
BMC Sports Science, Medicine &... Jul 2021The fact that kinesio tape may be capable to enhance muscle power would qualify it as practical tool to be considered during passive warm-up (WU) or coupled with active...
BACKGROUND
The fact that kinesio tape may be capable to enhance muscle power would qualify it as practical tool to be considered during passive warm-up (WU) or coupled with active WU processes prior to power-based performance. Therefore, the aim of this study was to investigate the single and combined effect of kinesio tape (KT) and WU on sprint cycling performance.
METHODS
In a repeated measure design, fifteen participants underwent six sessions to assess sprint cycling performance involving a combination of three taping conditions (without KT: NoKT; with KT positioned vertically over the thigh muscles KT; with KT positioned horizontally over the thigh muscles: Sham) with two pre-exercise routines (with WU: WU; without WU: NoWU) in a randomized order. Allometric scaling of peak power (PP) and average power (AP) values were considered for each sprint.
RESULTS
KT-WU demonstrated the highest PP and AP with respect to the other conditions (p < 0.05), except for AP that was similar to Sham-WU (p > 0.05). Moreover, NoKT-NoWU showed the lowest PP and AP with respect to the other conditions (p < 0.05).
CONCLUSIONS
Overall, our findings suggest that kinesio tape might be a possible tool to be combined with an active WU routine, inducing benefit on sprint performance. Moreover, KT may be considered a potential strategy to include within a passive WU, perhaps where an active WU is not feasible. However, as the influence of KT on muscle function is still unclear, our results should not be overinterpreted.
PubMed: 34311773
DOI: 10.1186/s13102-021-00310-3 -
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 -
Biosensors Dec 2021Conventional sample preparation techniques require bulky and expensive instruments and are not compatible with next-generation point-of-care diagnostic testing. Here, we...
Conventional sample preparation techniques require bulky and expensive instruments and are not compatible with next-generation point-of-care diagnostic testing. Here, we report a manually operated syringe-tip inertial microfluidic centrifuge (named i-centrifuge) for high-flow-rate (up to 16 mL/min) cell concentration and experimentally demonstrate its working mechanism and performance. Low-cost polymer films and double-sided tape were used through a rapid nonclean-room process of laser cutting and lamination bonding to construct the key components of the i-centrifuge, which consists of a syringe-tip flow stabilizer and a four-channel paralleled inertial microfluidic concentrator. The unstable liquid flow generated by the manual syringe was regulated and stabilized with the flow stabilizer to power inertial focusing in a four-channel paralleled concentrator. Finally, we successfully used our i-centrifuge for manually operated cell concentration. This i-centrifuge offers the advantages of low device cost, simple hand-powered operation, high-flow-rate processing, and portable device volume. Therefore, it holds potential as a low-cost, portable sample preparation tool for point-of-care diagnostic testing.
Topics: Microfluidic Analytical Techniques; Microfluidics; Polymers; Syringes
PubMed: 35049644
DOI: 10.3390/bios12010014 -
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