Anatomically and Functionally Distinct Lung Mesenchymal Populations Marked by Lgr5 and Lgr6.

The diversity of mesenchymal cell types in the lung that influence epithelial homeostasis and regeneration is poorly defined. We used genetic lineage tracing, single-cell RNA sequencing, and organoid culture approaches to show that Lgr5 and Lgr6, well-known markers of stem cells in epithelial tissues, are markers of mesenchymal cells in the adult lung. Lgr6+ cells comprise a subpopulation of smooth muscle cells surrounding airway epithelia and promote airway differentiation of epithelial progenitors via Wnt-Fgf10 cooperation. Genetic ablation of Lgr6+ cells impairs airway injury repair in vivo. Distinct Lgr5+ cells are located in alveolar compartments and are sufficient to promote alveolar differentiation of epithelial progenitors through Wnt activation. Modulating Wnt activity altered differentiation outcomes specified by mesenchymal cells. This identification of region- and lineage-specific crosstalk between epithelium and their neighboring mesenchymal partners provides new understanding of how different cell types are maintained in the adult lung.

Lung stem cell differentiation in mice directed by endothelial cells via a BMP4-NFATc1-thrombospondin-1 axis.

Lung stem cells are instructed to produce lineage-specific progeny through unknown factors in their microenvironment. We used clonal 3D cocultures of endothelial cells and distal lung stem cells, bronchioalveolar stem cells (BASCs), to probe the instructive mechanisms. Single BASCs had bronchiolar and alveolar differentiation potential in lung endothelial cell cocultures. Gain- and loss-of-function experiments showed that BMP4-Bmpr1a signaling triggers calcineurin/NFATc1-dependent expression of thrombospondin-1 (Tsp1) in lung endothelial cells to drive alveolar lineage-specific BASC differentiation. Tsp1 null mice exhibited defective alveolar injury repair, confirming a crucial role for the BMP4-NFATc1-TSP1 axis in lung epithelial differentiation and regeneration in vivo. Discovery of this pathway points to methods to direct the derivation of specific lung epithelial lineages from multipotent cells. These findings elucidate a pathway that may be a critical target in lung diseases and provide tools to understand the mechanisms of respiratory diseases at the single-cell level.

Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity.

The SARS-CoV-2 Omicron BA.1 variant emerged in 20211 and has multiple mutations in its spike protein2. Here we show that the spike protein of Omicron has a higher affinity for ACE2 compared with Delta, and a marked change in its antigenicity increases Omicron's evasion of therapeutic monoclonal and vaccine-elicited polyclonal neutralizing antibodies after two doses. mRNA vaccination as a third vaccine dose rescues and broadens neutralization. Importantly, the antiviral drugs remdesivir and molnupiravir retain efficacy against Omicron BA.1. Replication was similar for Omicron and Delta virus isolates in human nasal epithelial cultures. However, in lung cells and gut cells, Omicron demonstrated lower replication. Omicron spike protein was less efficiently cleaved compared with Delta. The differences in replication were mapped to the entry efficiency of the virus on the basis of spike-pseudotyped virus assays. The defect in entry of Omicron pseudotyped virus to specific cell types effectively correlated with higher cellular RNA expression of TMPRSS2, and deletion of TMPRSS2 affected Delta entry to a greater extent than Omicron. Furthermore, drug inhibitors targeting specific entry pathways3 demonstrated that the Omicron spike inefficiently uses the cellular protease TMPRSS2, which promotes cell entry through plasma membrane fusion, with greater dependency on cell entry through the endocytic pathway. Consistent with suboptimal S1/S2 cleavage and inability to use TMPRSS2, syncytium formation by the Omicron spike was substantially impaired compared with the Delta spike. The less efficient spike cleavage of Omicron at S1/S2 is associated with a shift in cellular tropism away from TMPRSS2-expressing cells, with implications for altered pathogenesis.

Tracing oncogene-driven remodelling of the intestinal stem cell niche.

Interactions between tumour cells and the surrounding microenvironment contribute to tumour progression, metastasis and recurrence1-3. Although mosaic analyses in Drosophila have advanced our understanding of such interactions4,5, it has been difficult to engineer parallel approaches in vertebrates. Here we present an oncogene-associated, multicolour reporter mouse model-the Red2Onco system-that allows differential tracing of mutant and wild-type cells in the same tissue. By applying this system to the small intestine, we show that oncogene-expressing mutant crypts alter the cellular organization of neighbouring wild-type crypts, thereby driving accelerated clonal drift. Crypts that express oncogenic KRAS or PI3K secrete BMP ligands that suppress local stem cell activity, while changes in PDGFRloCD81+ stromal cells induced by crypts with oncogenic PI3K alter the WNT signalling environment. Together, these results show how oncogene-driven paracrine remodelling creates a niche environment that is detrimental to the maintenance of wild-type tissue, promoting field transformation dominated by oncogenic clones.

SARS-CoV-2 B.1.617.2 Delta variant replication and immune evasion.

The B.1.617.2 (Delta) variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in the state of Maharashtra in late 2020 and spread throughout India, outcompeting pre-existing lineages including B.1.617.1 (Kappa) and B.1.1.7 (Alpha)1. In vitro, B.1.617.2 is sixfold less sensitive to serum neutralizing antibodies from recovered individuals, and eightfold less sensitive to vaccine-elicited antibodies, compared with wild-type Wuhan-1 bearing D614G. Serum neutralizing titres against B.1.617.2 were lower in ChAdOx1 vaccinees than in BNT162b2 vaccinees. B.1.617.2 spike pseudotyped viruses exhibited compromised sensitivity to monoclonal antibodies to the receptor-binding domain and the amino-terminal domain. B.1.617.2 demonstrated higher replication efficiency than B.1.1.7 in both airway organoid and human airway epithelial systems, associated with B.1.617.2 spike being in a predominantly cleaved state compared with B.1.1.7 spike. The B.1.617.2 spike protein was able to mediate highly efficient syncytium formation that was less sensitive to inhibition by neutralizing antibody, compared with that of wild-type spike. We also observed that B.1.617.2 had higher replication and spike-mediated entry than B.1.617.1, potentially explaining the B.1.617.2 dominance. In an analysis of more than 130 SARS-CoV-2-infected health care workers across three centres in India during a period of mixed lineage circulation, we observed reduced ChAdOx1 vaccine effectiveness against B.1.617.2 relative to non-B.1.617.2, with the caveat of possible residual confounding. Compromised vaccine efficacy against the highly fit and immune-evasive B.1.617.2 Delta variant warrants continued infection control measures in the post-vaccination era.

Airway secretory cell fate conversion via YAP-mTORC1-dependent essential amino acid metabolism.

Tissue homeostasis requires lineage fidelity of stem cells. Dysregulation of cell fate specification and differentiation leads to various diseases, yet the cellular and molecular mechanisms governing these processes remain elusive. We demonstrate that YAP/TAZ activation reprograms airway secretory cells, which subsequently lose their cellular identity and acquire squamous alveolar type 1 (AT1) fate in the lung. This cell fate conversion is mediated via distinctive transitional cell states of damage-associated transient progenitors (DATPs), recently shown to emerge during injury repair in mouse and human lungs. We further describe a YAP/TAZ signaling cascade to be integral for the fate conversion of secretory cells into AT1 fate, by modulating mTORC1/ATF4-mediated amino acid metabolism in vivo. Importantly, we observed aberrant activation of the YAP/TAZ-mTORC1-ATF4 axis in the altered airway epithelium of bronchiolitis obliterans syndrome, including substantial emergence of DATPs and AT1 cells with severe pulmonary fibrosis. Genetic and pharmacologic inhibition of mTORC1 activity suppresses lineage alteration and subepithelial fibrosis driven by YAP/TAZ activation, proposing a potential therapeutic target for human fibrotic lung diseases.

Author Correction: Metastatic-niche labelling reveals parenchymal cells with stem features.

An Amendment to this paper has been published and can be accessed via a link at the top of the paper.

Metastatic-niche labelling reveals parenchymal cells with stem features.

Direct investigation of the early cellular changes induced by metastatic cells within the surrounding tissue remains a challenge. Here we present a system in which metastatic cancer cells release a cell-penetrating fluorescent protein, which is taken up by neighbouring cells and enables spatial identification of the local metastatic cellular environment. Using this system, tissue cells with low representation in the metastatic niche can be identified and characterized within the bulk tissue. To highlight its potential, we applied this strategy to study the cellular environment of metastatic breast cancer cells in the lung. We report the presence of cancer-associated parenchymal cells, which exhibit stem-cell-like features, expression of lung progenitor markers, multi-lineage differentiation potential and self-renewal activity. In ex vivo assays, lung epithelial cells acquire a cancer-associated parenchymal-cell-like phenotype when co-cultured with cancer cells and support their growth. These results highlight the potential of this method as a platform for new discoveries.

Wearable Fabric Loop Sensor Based on Magnetic-Field-Induced Conductivity for Simultaneous Detection of Cardiac Activity and Respiration Signals.

In this study, a noncontact fabric loop sensor based on magnetic-field-induced conductivity, which can simultaneously detect cardiac activity and respiration signals, was developed and the effects of the sensor's shape and measurement position on the sensing performance were analyzed. Fifteen male subjects in their twenties wore sleeveless shirts equipped with various types of fabric loop sensors (spiky, extrusion, and spiral), and the cardiac activity and respiratory signals were measured twice at positions P2, P4, and P6. The measurements were verified by comparing them against the reference electrocardiogram (ECG) and respiratory signals measured using BIOPAC® (MP150, ECG100B, RSP100C). The waveforms of the raw signal measured by the fabric loop sensor were filtered with a bandpass filter (1-20 Hz) and qualitatively compared with the ECG signal obtained from the Ag/AgCI electrode. Notwithstanding a slight difference in performance, the three fabric sensors could simultaneously detect cardiac activity and respiration signals at all measurement positions. In addition, it was verified through statistical analysis that the highest-quality signal was obtained at the measurement position of P4 or P6 using the spiral loop sensor.

Machine Embroidered Sensors for Limb Joint Movement-Monitoring Smart Clothing.

In this study, a strain gauge sensor based on a change of contact or network structure between conductive materials was implemented using the handle-machine embroidery technique, and the variables (embroidery shape, embroidery distance, embroidery size, and implementation location) affecting its performance were studied. As a result of Experiment I on the structure of embroidery suitable for joint motion monitoring, the embroidery distance, rather than the embroidery size, was found to have a significant effect on the electric resistance changes caused by elongation. Based on the results of Experiment I, two types of zigzag embroideries, four types of embroideries with few contact points, and two types of embroideries with more contact points (all with short distances (2.0)) were selected for Experiment II (the dummy motion experiment). As a result of the dummy motion experiment, it was found that the locations of the suitable embroidered sensors for joint motion monitoring was the HJP (Hinge Joint Position) in the 'types without a contact point' (zigzag) and the LHJP (Lower Hinge Joint Position) in the 'types with more contact points'. On the other hand, although there was no consistency among the 'types with few contact points', the resistance changes measured by the 2CP and 7CP embroidered sensors showed similar figures and patterns, and the HJP location was most suitable. The resistance changes measured by the 4CP and 6CP embroidered sensors exhibited no consistent patterns, but the LHJP locations were more suitable. These results indicate that the location of the HJP is suitable for measuring joint motion in the 'type without a contact point', and the location of the LHJP is suitable for measuring joint motion when the number of contact points exceeds a certain limit. Among them, the average resistance change of the 9CP sensor located at the LHJP was 40 Ω with the smallest standard deviation of less than 1, and it is thus considered to have the best performance among all the sensors.

Conditions for CNT - Coated Textile Sensors Applied to Wearable Platforms to Monitor Limb Joint Motion.

Despite recent research on joint motion measurement to monitor human body movement, current measurement techniques and tools have significant limitations, including requiring large space for measurement and causing discomfort in test subjects wearing motion sensors. Our study aims, first, to develop carbon nanotube (CNT)-based textile joint motion sensors. Second, ours study aims to identify the most suitable CNT-based sensor structure and attachment method for use on a wearable platform during general exercise speeds. Lastly, we used these sensors on the human body, using sleeves and legs to find the most stable location, and we used the CNT-based sensor condition to monitor joint motions. We utilized our CNT-based sensor, which has proper elasticity as well as conductivity, and applied it to the elbow and knee joints. Based on the strain gauge principle, we monitored the variance of electric resistance that occurred when the CNT-based sensor was stretched due to limb motion. Our study tested 48 types of sensors. These sensors were applied to the CNT using different base knit textiles as well as different attachment methods, layers, sensor lengths, and sensor widths. The four most successful sensor types, which showed superior efficacy over the others in joint motion measurement, were selected for further study. These four sensors were then used to measure the elbow and knee joint motions of human subjects by placing them on different locations on sleeves and legs. The CNT knit textile sensors best suited to measuring joint motions are those with a double-layered CNT knit and 5 cm long × 0.5 cm or 1 cm wide sensors attached to a polyester¬-based knit using a welding method. The best position for the sensor to more stably monitor joint motions was the "below hinge position" from the elbow or knee hinge joint. Our study suggests an alternative strategy for joint-motion measurement that could contribute to the development of more comfortable and human-friendly methods of human limb motion measurement.

Stunning symmetries involved in the self-assembly of the HSV-1 capsid.

Herpes simplex virus-1 (HSV-1) is an enveloped dsDNA virus, infecting ~ 67% of humans. Here, we present the essential components of the HSV-1, focusing on stunning symmetries on the capsid. However, little is known about how the symmetries are involved dynamically in the self-assembly process. We suggest small angle X-ray scattering as a suitable method to capture the dynamics of self-assembly. Furthermore, our understanding of the viruses can be expanded by using an integrative approach that combines heterogeneous types of data, thus promoting new diagnostic tools and a cure for viral infections.

Evaluation of Joint Motion Sensing Efficiency According to the Implementation Method of SWCNT-Coated Fabric Motion Sensor.

The purpose of this study was to investigate the effects of the shape and attachment position of stretchable textile piezoresistive sensors coated with single-walled carbon nanotubes on their performance in measuring the joint movements of children. The requirements for fabric motion sensors suitable for children are also identified. The child subjects were instructed to wear integrated clothing with sensors of different shapes (rectangular and boat-shaped), attachment positions (at the knee and elbow joints or 4 cm below the joints). The change in voltage caused by the elongation and contraction of the fabric sensors was measured for the flexion-extension motions of the arms and legs at 60°/s (three measurements of 10 repetitions each for the 60° and 90° angles, for a total of 60 repetitions). Their reliability was verified by analyzing the agreement between the fabric motion sensors and attached acceleration sensors. The experimental results showed that the fabric motion sensor that can measure children's arm and leg motions most effectively is the rectangular-shaped sensor attached 4 cm below the joint. In this study, we developed a textile piezoresistive sensor suitable for measuring the joint motion of children, and analyzed the shape and attachment position of the sensor on clothing suitable for motion sensing. We showed that it is possible to sense joint motions of the human body by using flexible fabric sensors integrated into clothing.

Hybrid LPG-FBG Based High-Resolution Micro Bending Strain Sensor.

Sensitivity and reliability are essential factors for the practical implementation of a wearable sensor. This study explores the possibility of using a hybrid high-resolution Bragg grating sensor for achieving a fast response to dynamic, continuous motion and Bragg signal pattern monitoring measurement. The wavelength shift pattern for real-time monitoring in picometer units was derived by using femtosecond laser Bragg grating processing on an optical wave path with long-period grating. The possibility of measuring the demodulation system's Bragg signal pattern on the reflection spectrum of the femtosecond laser precision Bragg process and the long-period grating was confirmed. By demonstrating a practical method of wearing the sensor, the application of wearables was also explored. It is possible to present the applicability of sophisticated micro transformation measurement applications in picometer units.

Developmental mechanisms and adult stem cells for therapeutic lung regeneration.

Chronic degenerative lung diseases are essentially untreatable pathological conditions. By contrast, the healthy lung has numerous mechanisms that allow for rapid repair and restoration of function following minor acute injuries. We discuss the normal endogenous processes of lung development, homeostatic maintenance and repair and consider the research strategies required for the development of methods for human therapeutic lung regeneration.

Organogenesis of adult lung in a dish: Differentiation, disease and therapy.

The remarkable regenerative capacity of the lung suggests that stem cells could be of therapeutic importance in diverse lung diseases; however, the successful exploitation of lung stem cell biology has long been hampered by our inability to maintain and expand adult lung stem cells while retaining their multi-lineage potential in vitro. Recently, advances in our understanding of stem cell niches and the role of key signalling modulators in controlling stem cell maintenance and differentiation have fuelled the development of new in vitro three-dimensional (3D) culture technologies that sustain the stem cell-driven formation of near-physiological, self-organizing structures called organoids. Here we review basic approaches to organoid model systems and highlight recent achievements in the generation of organoids from adult stem and progenitor cells of both the murine and human lungs. We evaluate current applications in studying cellular changes in proliferation, differentiation, plasticity, and cell polarity, and cellular and molecular crosstalk of epithelial cells with stroma. Advantages and limitations of organoids for clinical use are also discussed.

Formation and magnetic properties of InFeP:Ag nanorods fabricated with noble metal Ag using an ion milling method.

The formation, including the density and height of the InFeP:Ag nanorods doped with noble metal Ag using an ion milling method, was preponderantly determined from transmission electron microscopy and x-ray diffraction analyses. We investigate, in particular, the enhanced ferromagnetism of the well-aligned InFeP:Ag nanorods. Auger electron spectroscopy and x-ray photoelectron spectroscopy measurements were carried out in order to investigate the incorporation of Ag and to verify the local chemical bonding of the InFeP:Ag nanorods. The variation of FWHM for the double-crystal x-ray rocking curve and triple-axis diffraction peaks demonstrates that noble metal Ag is incorporated into the InFeP:Ag nanorods. The noticeable ferromagnetic signature (M-H curve) of the InFeP:Ag nanorods is observed and T c persists up to almost 350 K (3.9 × 10-4 emu g-1), as determined by temperature-dependence magnetization (M-T curve) measurements. This study suggests that the InFeP:Ag nanorods should be a potential candidate for the application of spintronic devices.

IL-13 Augments Compressive Stress-Induced Tissue Factor Expression in Human Airway Epithelial Cells.

Tissue factor (TF) is best known as a cellular initiator of coagulation, but it is also a multifunctional protein that has been implicated in multiple pathophysiologic conditions, including asthma. In the lung, airway epithelial cells express TF, but it is unknown how TF expression is regulated by asthma-associated mediators. We investigated the role of IL-13, a type 2 cytokine, alone and in combination with compressive stress, which mimics asthmatic bronchoconstriction, on TF expression and release of TF-positive extracellular vesicles from primary normal human bronchial epithelial cells. Well-differentiated normal human bronchial epithelial cells were treated with IL-13 and compressive stress, alone and in combination. TF mRNA, protein and activity were measured in the cells and conditioned media. TF was also measured in the bronchoalveolar lavage (BAL) fluid of allergen-challenged mice and patients with asthma. IL-13 and compressive stress increased TF expression, but only compressive stress induced TF-positive extracellular vesicle release. Pretreatment with IL-13 augmented compressive stress-induced TF expression and release. TF protein and activity in BAL fluid were increased in allergen-sensitized and -challenged mice. TF was elevated in the BAL fluid of patients with mild asthma after an allergen challenge. Our in vitro and in vivo data indicate close cooperation between mechanical and inflammatory stimuli on TF expression and release of TF-positive extracellular vesicles in the lungs, which may contribute to pathophysiology of asthma.

A Study on the Optimal Positions of ECG Electrodes in a Garment for the Design of ECG-Monitoring Clothing for Male.

Smart clothing is a sort of wearable device used for ubiquitous health monitoring. It provides comfort and efficiency in vital sign measurements and has been studied and developed in various types of monitoring platforms such as T-shirt and sports bra. However, despite these previous approaches, smart clothing for electrocardiography (ECG) monitoring has encountered a serious shortcoming relevant to motion artifacts caused by wearer movement. In effect, motion artifacts are one of the major problems in practical implementation of most wearable health-monitoring devices. In the ECG measurements collected by a garment, motion artifacts are usually caused by improper location of the electrode, leading to lack of contact between the electrode and skin with body motion. The aim of this study was to suggest a design for ECG-monitoring clothing contributing to reduction of motion artifacts. Based on the clothing science theory, it was assumed in this study that the stability of the electrode in a dynamic state differed depending on the electrode location in an ECG-monitoring garment. Founded on this assumption, effects of 56 electrode positions were determined by sectioning the surface of the garment into grids with 6 cm intervals in the front and back of the bodice. In order to determine the optimal locations of the ECG electrodes from the 56 positions, ECG measurements were collected from 10 participants at every electrode position in the garment while the wearer was in motion. The electrode locations indicating both an ECG measurement rate higher than 80.0 % and a large amplitude during motion were selected as the optimal electrode locations. The results of this analysis show four electrode locations with consistently higher ECG measurement rates and larger amplitudes amongst the 56 locations. These four locations were abstracted to be least affected by wearer movement in this research. Based on this result, a design of the garment-formed ECG monitoring platform reflecting the optimal positions of the electrode was suggested.

The Effect of Electrode Designs Based on the Anatomical Heart Location for the Non-Contact Heart Activity Measurement.

This research is an extension of a previous research [1] on the different effects of sensor location that is relatively suitable for heart rate sensing. This research aimed to elucidate the causes of wide variations in heart rate measurements from the same sensor position among subjects, as observed in previous research [1], and to enhance designs of the inductive textile electrode to overcome these variations. To achieve this, this study comprised two parts: In part 1, X-ray examinations were performed to determine the cause of the wide variations noted in the findings from previous research [1], and we found that at the same sensor position, the heart activity signal differed with slight differences in the positions of the heart of each subject owing to individual differences in the anatomical heart location. In part 2, three types of dual-loop-type textile electrodes were devised to overcome variations in heart location that were confirmed in part 1 of the study. The variations with three types of sensor designs were compared with that with a single-round type of electrode design, by using computer simulation and by performing a t-test on the data obtained from the experiments. We found that the oval-oval shaped, dual-loop-type textile electrode was more suitable than the single round type for determining morphological characteristics as well as for measuring appropriate heart activity signals. Based on these results, the oval-oval, dual-loop-type was a better inductive textile electrode that more effectively overcomes individual differences in heart location during heart activity sensing based on the magnetic-induced conductivity principle.