The novel prognostic marker SPOCK2 regulates tumour progression in melanoma.

Secreted protein acidic and cysteine rich/osteonectin, cwcv and kazal-like domain proteoglycan 2 (SPOCK2) is a protein that regulates cell differentiation and growth. Recent studies have reported that SPOCK2 plays important roles in the progression of various human cancers; however, the role of SPOCK2 in melanoma remains unknown. Therefore, this study investigated the roles of SPOCK2 and the related mechanisms in melanoma progression. To evaluate the clinical significance of SPOCK2 expression in patients with melanoma, we analysed the association between SPOCK2 expression and its prognostic value for patients with melanoma using systematic multiomic analysis. Subsequently, to investigate the roles of Spock2 in melanoma progression in vitro and in vivo, we knocked down Spock2 in the B16F10 melanoma cell line. High SPOCK2 levels were positively associated with good prognosis and long survival rate of patients with melanoma. Spock2 knockdown promoted melanoma cell proliferation by inducing the cell cycle and inhibiting apoptosis. Moreover, Spock2 downregulation significantly increased cell migration and invasion by upregulating MMP2 and MT1-MMP. The increased cell proliferation and migration were inhibited by MAPK inhibitor, and ERK phosphorylation was considerably enhanced in Spock2 knockdown cells. Therefore, Spock2 could function as a tumour suppressor gene to regulate melanoma progression by regulating the MAPK/ERK signalling pathway. Additionally, Spock2 knockdown cell injection induced considerable tumour growth and lung metastasis in C57BL6 mice compared to that in the control group. Our findings suggest that SPOCK2 plays crucial roles in malignant progression of melanoma and functions as a novel therapeutic target of melanoma.

Sizzled (Frzb3) physically interacts with noncanonical Wnt ligands to inhibit gastrulation cell movement.

The coordinated movement of germ layer progenitor cells reaches its peak at the dorsal side, where the Bmp signaling gradient is low, and minimum at the ventral side, where the Bmp gradient is high. This dynamic cell movement is regulated by the interplay of various signaling pathways. The noncanonical Wnt signaling cascade serves as a pivotal regulator of convergence and extension cell movement, facilitated by the activation of small GTPases such as Rho, Rab, and Rac. However, the underlying cause of limited cell movement at the ventral side remains elusive. To explore the functional role of a key regulator in constraining gastrulation cell movement at the ventral side, we investigated the Bmp4-direct target gene, sizzled (szl), to assess its potential role in inhibiting noncanonical Wnt signaling. In our current study, we demonstrated that ectopic expression of szl led to gastrulation defects in a dose-dependent manner without altering cell fate specification. Overexpression of szl resulted in decreased elongation of Activin-treated animal cap and Keller explants. Furthermore, our immunoprecipitation assay unveiled the physical interaction of Szl with noncanonical Wnt ligand proteins (Wnt5 and Wnt11). Additionally, the activation of small GTPases involved in Wnt signaling mediation (RhoA and Rac1) was diminished upon szl overexpression. In summary, our findings suggest that Bmp4 signaling negatively modulates cell movement from the ventral side of the embryo by inducing szl expression during early Xenopus gastrulation.

Deup1 Expression Interferes with Multiciliated Differentiation.

A recent study revealed that the loss of Deup1 expression does not affect either centriole amplification or multicilia formation. Therefore, the deuterosome per se is not a platform for amplification of centrioles. In this study, we examine whether gain-of-function of Deup1 affects the development of multiciliated ependymal cells. Our time-lapse study reveals that deuterosomes with an average diameter of 300 nm have two different fates during ependymal differentiation. In the first instance, deuterosomes are scattered and gradually disappear as cells become multiciliated. In the second instance, deuterosomes self-organize into a larger aggregate, called a deuterosome cluster (DC). Unlike scattered deuterosomes, DCs possess centriole components primarily within their large structure. A characteristic of DC-containing cells is that they tend to become primary ciliated rather than multiciliated. Our in utero electroporation study shows that DCs in ependymal tissue are mostly observed at early postnatal stages, but are scarce at late postnatal stages, suggesting the presence of DC antagonists within the differentiating cells. Importantly, from our bead flow assay, ectopic expression of Deup1 significantly impairs cerebrospinal fluid flow. Furthermore, we show that expression of mouse Deup1 in Xenopus embryos has an inhibitory effect on differentiation of multiciliated cells in the epidermis. Taken together, we conclude that the DC formation of Deup1 in multiciliated cells inhibits production of multiple centrioles.

ANKS1A-Deficiency Aberrantly Increases the Entry of the Protein Transport Machinery into the Ependymal Cilia.

In this study, we examine whether a change in the protein levels for FOP in Ankyrin repeat and SAM domain-containing protein 1A (ANKS1A)-deficient ependymal cells affects the intraflagellar transport (IFT) protein transport system in the multicilia. Three distinct abnormalities are observed in the multicilia of ANKS1A-deficient ependymal cells. First, there were a greater number of IFT88-positive trains along the cilia from ANKS1A deficiency. The results are similar to each isolated cilium as well. Second, each isolated cilium contains a significant increase in the number of extracellular vesicles (ECVs) due to the lack of ANKS1A. Third, Van Gogh-like 2 (Vangl2), a ciliary membrane protein, is abundantly detected along the cilia and in the ECVs attached to them for ANKS1A-deficient cells. We also use primary ependymal culture systems to obtain the ECVs released from the multicilia. Consequently, we find that ECVs from ANKS1A-deficient cells contain more IFT machinery and Vangl2. These results indicate that ANKS1A deficiency increases the entry of the protein transport machinery into the multicilia and as a result of these abnormal protein transports, excessive ECVs form along the cilia. We conclude that ependymal cells make use of the ECV-based disposal system in order to eliminate excessively transported proteins from basal bodies.

ANKS1A regulates LDL receptor-related protein 1 (LRP1)-mediated cerebrovascular clearance in brain endothelial cells.

Brain endothelial LDL receptor-related protein 1 (LRP1) is involved in the clearance of Aß peptides across the blood-brain barrier (BBB). Here we show that endothelial deficiency of ankyrin repeat and SAM domain containing 1 A (ANKS1A) reduces both the cell surface levels of LRP1 and the Aß clearance across the BBB. Association of ANKS1A with the NPXY motifs of LRP1 facilitates the transport of LRP1 from the endoplasmic reticulum toward the cell surface. ANKS1A deficiency in an Alzheimer's disease mouse model results in exacerbated Aß pathology followed by cognitive impairments. These deficits are reversible by gene therapy with brain endothelial-specific ANKS1A. In addition, human induced pluripotent stem cell-derived BBBs (iBBBs) were generated from endothelial cells lacking ANKS1A or carrying the rs6930932 variant. Those iBBBs exhibit both reduced cell surface LRP1 and impaired Aß clearance. Thus, our findings demonstrate that ANKS1A regulates LRP1-mediated Aß clearance across the BBB.

Plant cell-like tip-growing polymer precipitate with structurally embedded multistimuli sensing ability.

Soft systems that respond to external stimuli, such as heat, magnetic field, and light, find applications in a range of fields including soft robotics, energy harvesting, and biomedicine. However, most of the existing systems exhibit nondirectional, nastic movement as they can neither grow nor sense the direction of stimuli. In this regard, artificial systems are outperformed by organisms capable of directional growth in response to the sense of stimuli or tropic growth. Inspired by tropic growth schemes of plant cells and fungal hyphae, here we report an artificial multistimuli-responsive tropic tip-growing system based on nonsolvent-induced phase separation of polymer solution, where polymer precipitates as its solvent dissolves into surrounding nonsolvent. We provide a theoretical framework to predict the size and velocity of growing precipitates and demonstrate its capability of sensing the directions of gravity, mechanical contact, and light and adjusting its growing direction in response. Exploiting the embedded physical intelligence of sensing and responding to external stimuli, our soft material system achieves multiple tasks including printing 3D structures in a confined space, bypassing mechanical obstacles, and shielded transport of liquids within water.

Cerebral Cavernous Malformation (CCM)-like Vessel Lesion in the Aged ANKS1A-deficient Brain.

In this study, we show that ANKS1A is specifically expressed in the brain endothelial cells of adult mice. ANKS1A deficiency in adult mice does not affect the differentiation, growth, or patterning of the cerebrovascular system; however, its absence significantly impacts the cerebrovascular system of the aged brain. In aged ANKS1A knock-out (KO) brains, vessel lesions exhibiting cerebral cavernous malformations (CCMs) are observed. In addition, CCM-like lesions show localized peripheral blood leakage into the brain. The CCM-like lesions reveal immune cells infiltrating the parenchyma. The CCM-like lesions also contain significantly fewer astrocyte endfeets and tight junctions, indicating that the integrity of the BBB has been partially compromised. CCM-like lesions display increased fibronectin expression in blood vessels, which is also confirmed in cultured endothelial cells deficient for ANKS1A. Therefore, we hypothesize that ANKS1A may play a role in maintaining or stabilizing healthy blood vessels in the brain during aging.

Systematic Multiomic Analysis of PKHD1L1 Gene Expression and Its Role as a Predicting Biomarker for Immune Cell Infiltration in Skin Cutaneous Melanoma and Lung Adenocarcinoma.

The identification of genetic factors that regulate the cancer immune microenvironment is important for understanding the mechanism of tumor progression and establishing an effective treatment strategy. Polycystic kidney and hepatic disease 1-like protein 1 (PKHD1L1) is a large transmembrane protein that is highly expressed in immune cells; however, its association with tumor progression remains unclear. Here, we systematically analyzed the clinical relevance of PKHD1L1 in the tumor microenvironment in multiple cancer types using various bioinformatic tools. We found that the PKHD1L1 mRNA expression levels were significantly lower in skin cutaneous melanoma (SKCM) and lung adenocarcinoma (LUAD) than in normal tissues. The decreased expression of PKHD1L1 was significantly associated with unfavorable overall survival (OS) in SKCM and LUAD. Additionally, PKHD1L1 expression was positively correlated with the levels of infiltrating B cells, cluster of differentiation (CD)-8+ T cells, and natural killer (NK) cells, suggesting that the infiltration of immune cells could be associated with a good prognosis due to increased PKHD1L1 expression. Gene ontology (GO) analysis also revealed the relationship between PKHD1L1-co-altered genes and the activation of lymphocytes, including B and T cells. Collectively, this study shows that PKHD1L1 expression is positively correlated with a good prognosis via the induction of immune infiltration, suggesting that PKHD1L1 has potential prognostic value in SKCM and LUAD.

Perceived Safety and Pedestrian Performance in Pedestrian Priority Streets (PPSs) in Seoul, Korea: A Virtual Reality Experiment and Trace Mapping.

Pedestrian Priority Street (PPS) project, launched to encourage safer and more convenient walking by improving the inferior pedestrian environment on narrow streets without sidewalks, is based on Monderman's shared space concept. Similar to the shared space approach, PPS aims for mutual consideration between pedestrians and drivers and strives to create a pedestrian-friendly environment, but the project relies on a unique road surface design. Considering the two main goals of the PPS project, this study investigated how subjective safety and pedestrians' movements differed by design types. To analyze safety perception, ordered Logit regression and post-hoc interviews were conducted with visual assessment survey using recorded VR (virtual reality) videos. Next, trace mapping and analysis were performed based on the video recordings to measure the degree of free walking. The results found that pedestrians perceived higher safety level in PPSs than in general back road. Further, the pedestrians moved more freely in the street with an integrated design. In other types, which suggested a pedestrian zone at the roadside, there was not much difference in behavior from the general back roads. Thus, the design principle of PPS, which does not set a boundary between pedestrian and vehicle area, should be observed to lead to behavioral changes in pedestrians.

Hydrogel-Based Iontronics on a Polydimethylsiloxane Microchip.

In response to the extensive utilization of ionic circuits, including in iontronics and wearable devices, a new method for fabricating a hydrogel-based ionic circuit on a polydimethylsiloxane (PDMS) microchip is reported. Prolonged UV/ozone oxidation combined with proper surface functionalizations and a novel microchip bonding method using thiol-epoxy click reaction enable the robust attachment of the photopolymerized hydrogel to the microchannel surface for eventual operation in electrolytes as an ionic circuit. The stretchable ionic diode constructed on the PDMS microchip shows a superior rectification ratio even under tensile stress and long-term storage stability. Furthermore, the combination of the ionic circuit and unique material properties of PDMS allows us to maximize the versatility and diversify the functionalities of the iontronic device, as demonstrated in a pressure-driven ionic switch and chip-integrated ionic regulator. Its iontronic signal transmission mimicking the excitatory and inhibitory synapses also evinces the potential of the hydrogel-based iontronics on the PDMS microchip as developed toward an aqueous neuromimetic information processor while opening up new opportunities for various bioinspired applications.

The molecular dynamics of subdistal appendages in multi-ciliated cells.

The motile cilia of ependymal cells coordinate their beats to facilitate a forceful and directed flow of cerebrospinal fluid (CSF). Each cilium originates from a basal body with a basal foot protruding from one side. A uniform alignment of these basal feet is crucial for the coordination of ciliary beating. The process by which the basal foot originates from subdistal appendages of the basal body, however, is unresolved. Here, we show FGFR1 Oncogene Partner (FOP) is a useful marker for delineating the transformation of a circular, unpolarized subdistal appendage into a polarized structure with a basal foot. Ankyrin repeat and SAM domain-containing protein 1A (ANKS1A) interacts with FOP to assemble region I of the basal foot. Importantly, disruption of ANKS1A reduces the size of region I. This produces an unstable basal foot, which disrupts rotational polarity and the coordinated beating of cilia in young adult mice. ANKS1A deficiency also leads to severe degeneration of the basal foot in aged mice and the detachment of cilia from their basal bodies. This role of ANKS1A in the polarization of the basal foot is evolutionarily conserved in vertebrates. Thus, ANKS1A regulates FOP to build and maintain the polarity of subdistal appendages.

Chemically Robust Indium Tin Oxide/Graphene Anode for Efficient Perovskite Light-Emitting Diodes.

Graphene is an optimal material to be employed as an ionic diffusion barrier because of its outstanding impermeability and chemical robustness. Indium tin oxide (ITO) is often used in perovskite light-emitting diodes (PeLEDs), and it can release indium easily upon exposure to the acidic hole-injection layer so that luminescence can be quenched significantly. Here, we exploit the outstanding impermeability of graphene and use it as a chemical barrier to block the etching that can occur in ITO exposed to an acidic hole-injection layer in PeLEDs. This barrier reduced the luminescence quenching that these metallic species can cause, so the photoluminescence lifetime of perovskite film was substantially higher in devices with ITO and graphene layer (87.9 ns) than in devices that had only an ITO anode (22.1 ns). Luminous current efficiency was also higher in PeLEDs with a graphene barrier (16.4 cd/A) than in those without graphene (9.02 cd/A). Our work demonstrates that graphene can be used as a barrier to reduce the degradation of transparent electrodes by chemical etching in optoelectronic devices.

Reversible Crosslinking of Polymer/Metal-Ion Complexes for a Microfluidic Switch.

The importance of chitosan has been strongly emphasized in literature because this natural polymer could not only remove heavy metal ions in water but also have the potential for recyclability. However, reversible phase transition and its dynamics, which are highlighting areas of a recycle process, have not been studied sufficiently. Here, we present dynamic studies of the dissolution as well as the gelation of a physically crosslinked chitosan hydrogel. Specifically, a one-dimensional gel growth system and an acetate buffer solution were prepared for the precise analysis of the dominant factors affecting a phase transition. The dissolution rate was found to be regulated by three major factors of the pH level, Cu2+, and NO2 -, while the gelation rate was strongly governed by the concentration of OH-. Apart from the gelation rate, the use of Cu2+ led to the rapid realization of gel characteristics. The results here provide strategies for process engineering, ultimately to determine the phase-transition rates. In addition, a microfluidic switch was successfully operated based on a better understanding of the reversible crosslinking of the chitosan hydrogel. Rapid gelation was required to close the channel, and a quick switchover was achieved by a dissolution enhancement strategy. As a result, factors that regulated the rates of gelation or dissolution were found to be useful to operate the fluidic switch.

Ion-to-ion amplification through an open-junction ionic diode.

As biological signals are mainly based on ion transport, the differences in signal carriers have become a major issue for the intimate communication between electrical devices and biological areas. In this respect, an ionic device which can directly interpret ionic signals from biological systems needs to be designed. Particularly, it is also required to amplify the ionic signals for effective signal processing, since the amount of ions acquired from biological systems is very small. Here, we report the signal amplification in ionic systems as well as sensing through the modified design of polyelectrolyte hydrogel-based ionic diodes. By designing an open-junction structure, ionic signals from the external environment can be directly transmitted to an ionic diode. Moreover, the minute ionic signals injected into the devices can also be amplified to a large amount of ions. The signal transduction mechanism of the ion-to-ion amplification is suggested and clearly verified by revealing the generation of breakdown ionic currents during an ion injection. Subsequently, various methods for enhancing the amplification are suggested.

Ependymal Cells Require Anks1a for Their Proper Development.

Ependymal cells constitute the multi-ciliated epithelium, which lines the brain ventricular lumen. Although ependymal cells originate from radial glial cells in the perinatal rodent brain, the exact mechanisms underlying the full differentiation of ependymal cells are poorly understood. In this report, we present evidence that the Anks1a phosphotyrosine binding domain (PTB) adaptor is required for the proper development of ependymal cells in the rodent postnatal brain. Anks1a gene trap targeted LacZ reporter analysis revealed that Anks1a is expressed prominently in the ventricular region of the early postnatal brain and that its expression is restricted to mature ependymal cells during postnatal brain development. In addition, Anks1a-deficient ependymal cells were shown to possess type B cell characteristics, suggesting that ependymal cells require Anks1a in order to be fully differentiated. Finally, Anks1a overexpression in the lateral wall of the neonatal brain resulted in an increase in the number of ependymal cells during postnatal brain development. Altogether, our results suggest that ependymal cells require Anks1a PTB adaptor for their proper development.

Stretchable Ionics - A Promising Candidate for Upcoming Wearable Devices.

As many devices for human utility aim for fast and convenient communication with users, superb electronic devices are demonstrated to serve as hardware for human-machine interfaces in wearable forms. Wearable devices for daily healthcare and self-diagnosis offer more human-like properties unconstrained by deformation. In this sense, stretchable ionics based on flexible and stretchable hydrogels are on the rise as another means to develop wearable devices for bioapplications for two main reasons: i) ionic currents and choosing the same signal carriers for biological areas, and ii) the adoption of hydrogel ionic conductors, which are intrinsically stretchable materials with biocompatibility. Here, the current status of stretchable ionics and future applications are introduced, whose positive effects can be magnified by stretchable ionics.

Anks1a regulates COPII-mediated anterograde transport of receptor tyrosine kinases critical for tumorigenesis.

ErbB2 signalling, which is amplified by EphA2 binding, is an important therapeutic target for breast cancer. Despite the importance of the EphA2/ErbB2 complex in promoting breast tumorigenesis, the mechanism by which these receptor tyrosine kinases (RTKs) are exported from the endoplasmic reticulum (ER) remains poorly understood. Here we report that the PTB adaptor Anks1a is specifically localized to the ER on its own serine phosphorylation. Once there, Anks1a acts as an important regulator of COPII-mediated EphA2 ER export. The Anks1a ankyrin repeat domain binds EphA2 and causes it to accumulate at sites of ER exit. Simultaneously, the Anks1a PTB domain binds Sec23. This induces internalization of EphA2 via COPII vesicles, while Anks1a remains behind on the ER membrane. EphA2 also binds ErbB2 in the ER and seems to load ErbB2 into growing COPII carriers. Together, our study reveals a novel mechanism that regulates the loading of RTKs into COPII vesicles.

Proper closure of the optic fissure requires ephrin A5-EphB2-JNK signaling.

The development of complex organs such as the eye requires a delicate and coordinated balance of cell division and cell death. Although apoptosis is prevalent in the proximoventral optic cup, the precise role it plays in eye development needs to be investigated further. In this study, we show that reduced apoptosis in the proximoventral optic cup prevents closure of the optic fissure. We also show that expression of ephrin A5 (Efna5) partially overlaps with Eph receptor B2 (Ephb2) expression in the proximoventral optic cup and that binding of EphB2 to ephrin A5 induces a sustained activation of JNK. This prolonged JNK signal promotes apoptosis and prevents cell proliferation. Thus, we propose that the unique cross-subclass interaction of EphB2 with ephrin A5 has evolved to function upstream of JNK signaling for the purpose of maintaining an adequate pool of progenitor cells to ensure proper closure of the optic fissure.

Columnar grown copper films on polyimides strained beyond 100.

Many flexible electronic devices contain metal films on polymer substrates to satisfy requirements for both electrical conductivity and mechanical durability. Despite numerous trials to date, the stretchability of metal interconnects remains an issue. In this paper, we have demonstrated a stretchable metal interconnect through control of the texture of a copper film with columnar grown grains on a polyimide (PI) substrate. The columnar grown copper films (CGC films) were deposited by regulating radio frequency (RF) sputtering powers. CGC films were able to sustain their electrical conductivity at strains above 100%. Instead of ultimate electrical discontinuity by channel crack propagation, CGC films maintained their conductivity by forming ligament structures, or a 'conductive net,' through trapped micro-cracks. XRD, AFM and in situ SEM analysis were used to investigate these stretchable conductors.