Epitranscriptic regulation of HRAS by N6-methyladenosine drives tumor progression.

Overexpression of Ras, in addition to the oncogenic mutations, occurs in various human cancers. However, the mechanisms for epitranscriptic regulation of RAS in tumorigenesis remain unclear. Here, we report that the widespread N6-methyladenosine (m6A) modification of HRAS, but not KRAS and NRAS, is higher in cancer tissues compared with the adjacent tissues, which results in the increased expression of H-Ras protein, thus promoting cancer cell proliferation and metastasis. Mechanistically, three m6A modification sites of HRAS 3' UTR, which is regulated by FTO and bound by YTHDF1, but not YTHDF2 nor YTHDF3, promote its protein expression by the enhanced translational elongation. In addition, targeting HRAS m6A modification decreases cancer proliferation and metastasis. Clinically, up-regulated H-Ras expression correlates with down-regulated FTO and up-regulated YTHDF1 expression in various cancers. Collectively, our study reveals a linking between specific m6A modification sites of HRAS and tumor progression, which provides a new strategy to target oncogenic Ras signaling.

LCDR regulates the integrity of lysosomal membrane by hnRNP K-stabilized LAPTM5 transcript and promotes cell survival.

Lysosome plays important roles in cellular homeostasis, and its dysregulation contributes to tumor growth and survival. However, the understanding of regulation and the underlying mechanism of lysosome in cancer survival is incomplete. Here, we reveal a role for a histone acetylation-regulated long noncoding RNA termed lysosome cell death regulator (LCDR) in lung cancer cell survival, in which its knockdown promotes apoptosis. Mechanistically, LCDR binds to heterogenous nuclear ribonucleoprotein K (hnRNP K) to regulate the stability of the lysosomal-associated protein transmembrane 5 (LAPTM5) transcript that maintains the integrity of the lysosomal membrane. Knockdown of LCDR, hnRNP K, or LAPTM5 promotes lysosomal membrane permeabilization and lysosomal cell death, thus consequently resulting in apoptosis. LAPTM5 overexpression or cathepsin B inhibitor partially restores the effects of this axis on lysosomal cell death in vitro and in vivo. Similarly, targeting LCDR significantly decreased tumor growth of patient-derived xenografts of lung adenocarcinoma (LUAD) and had significant cell death using nanoparticles (NPs)-mediated systematic short interfering RNA delivery. Moreover, LCDR/hnRNP K/LAPTM5 are up-regulated in LUAD tissues, and coexpression of this axis shows the increased diagnostic value for LUAD. Collectively, we identified a long noncoding RNA that regulates lysosome function at the posttranscriptional level. These findings shed light on LCDR/hnRNP K/LAPTM5 as potential therapeutic targets, and targeting lysosome is a promising strategy in cancer treatment.

SNX27 suppresses SARS-CoV-2 infection by inhibiting viral lysosome/late endosome entry.

After binding to its cell surface receptor angiotensin converting enzyme 2 (ACE2), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the host cell through directly fusing with plasma membrane (cell surface pathway) or undergoing endocytosis traveling to lysosome/late endosome for membrane fusion (endocytic pathway). However, the endocytic entry regulation by host cell remains elusive. Recent studies show ACE2 possesses a type I PDZ binding motif (PBM) through which it could interact with a PDZ domain-containing protein such as sorting nexin 27 (SNX27). In this study, we determined the ACE2-PBM/SNX27-PDZ complex structure, and, through a series of functional analyses, we found SNX27 plays an important role in regulating the homeostasis of ACE2 receptor. More importantly, we demonstrated SNX27, together with retromer complex (the core component of the endosomal protein sorting machinery), prevents ACE2/virus complex from entering lysosome/late endosome, resulting in decreased viral entry in cells where the endocytic pathway dominates. The ACE2/virus retrieval mediated by SNX27-retromer could be considered as a countermeasure against invasion of ACE2 receptor-using SARS coronaviruses.

Hypoxia inhibits HUNK kinase activity to induce epithelial-mesenchymal transition.

Hypoxia is a common hallmark of cancer and plays a crucial role in promoting epithelial-mesenchymal transition (EMT). Hormonally Upregulated Neu-associated Kinase (HUNK) regulates EMT through its kinase activity. However, whether hypoxia is involved in HUNK-mediated EMT is incompletely understood. This study unveils an association between HUNK kinase activity and hypoxia in colorectal cancer (CRC). Importantly, hypoxia does not alter the expression levels of HUNK, but directly affects the phosphorylation levels of downstream proteins with indication of HUNK activity. Functionally, the upregulation of migration, invasion, and expression of EMT markers in CRC cells under hypoxic conditions can be attributed, in part, to the downregulation of HUNK-mediated phosphorylation of downstream proteins. These findings highlight the intricate relationship between HUNK, hypoxia and the molecular mechanisms of cancer EMT. Understanding these mechanisms may provide valuable insights into therapeutic targets for inhibiting cancer metastasis.

Tumor cell-intrinsic PD-1 receptor is a tumor suppressor and mediates resistance to PD-1 blockade therapy.

The programmed cell death 1 (PD-1) receptor on the surface of immune cells is an immune checkpoint molecule that mediates the immune escape of tumor cells. Consequently, antibodies targeting PD-1 have shown efficacy in enhancing the antitumor activity of T cells in some types of cancers. However, the potential effects of PD-1 on tumor cells remain largely unknown. Here, we show that PD-1 is expressed across a broad range of tumor cells. The silencing of PD-1 or its ligand, PD-1 ligand 1 (PD-L1), promotes cell proliferation and colony formation in vitro and tumor growth in vivo. Conversely, overexpression of PD-1 or PD-L1 inhibits tumor cell proliferation and colony formation. Moreover, blocking antibodies targeting PD-1 or PD-L1 promote tumor growth in cell cultures and xenografts. Mechanistically, the coordination of PD-1 and PD-L1 activates its major downstream signaling pathways including the AKT and ERK1/2 pathways, thus enhancing tumor cell growth. This study demonstrates that PD-1/PD-L1 is a potential tumor suppressor and potentially regulates the response to anti-PD-1/PD-L1 treatments, thus representing a potential biomarker for the optimal cancer immunotherapeutic treatment.

Conditioning of raw sludge and thermally hydrolyzed sludge by ferric salt and cationic polyacrylamide: rheological analysis.

In this study, the conditioning effect of cationic polyacrylamide (CPAM) with different charge densities on raw sludge (RS) and thermo-hydrolyzed sludge (HS) pretreated with or without ferric salt is studied through orthogonal experiments. In addition, this paper uses the principles of rheology and morphology to analyze and clarify the conditioning mechanism of RS and HS, and reveals the mechanism of thermal hydrolysis to improve the dewatering performance of sludge. Compared with the RS, the HS has smaller particle size, better filterability, stronger fluidity and more obvious thixotropy. However, due to the influence of filter pressing time, ferric salt should be added before conditioning. The orthogonal experiment shows that the optimal conditioner is CPAM with charge density of 60, and the specific resistance to filtration and capillary suction time of the adjusted thermo-hydrolyzed sludge are reduced to (1.11 ± 0.07) × 1012 m/kg and 16.1 ± 1.8 s; the particle size increased from 61.2 to 253.5 µm. The moisture content of the sludge cake is about 48%. The structural strength and thixotropy of HS are higher than those of the RS, and can be greatly improved by adding ferric salt. Morphological analysis confirms that thermal hydrolysis can lyse microbial cells in sludge, and the sludge treated with ferric salt will have more porous structure and stronger flocculation strength.

Comparative study on the association between types of physical activity, physical activity levels, and the incidence of osteoarthritis in adults: the NHANES 2007-2020.

It is known that physical activity is beneficial for the prevention of osteoarthritis (OA), but specific discussions on which types and levels of physical activity are more effective in reducing the incidence of OA are restricted. This study is aimed at exploring the correlation concerning the types of physical activity, levels of physical activity, and the incidence of OA by assessing the participation in five typical forms of physical activity (vigorous work activity, vigorous recreational activity, moderate work activity, moderate recreational activity, and walking or bicycling). Cross-sectional study was conducted. Self-reported data on specific types of physical activity were obtained from individuals in the National Health and Nutrition Examination Survey (NHANES) from 2007 to 2020 with the use of the Physical Activity Questionnaire (PAQ). The incidence of OA was assessed through the "Health Conditions" questionnaire section of NHANES. Weighted logistic regression analysis was employed to study the correlation between physical activity types and levels, and the incidence of OA. Different kinds of physical activity and physical activity levels have varying impacts on the incidence of OA. Among the types of physical activity, vigorous recreational activity and moderate recreational activity are found to have a preventive effect on OA. In terms of physical activity levels, low physical activity levels of moderate work activity are associated with an increased risk of OA, while moderate physical activity levels are confirmed to have a protective effect against OA in the age groups of 20-44 and 45-64. However, gender-stratified analyses reveal that both low and moderate physical activity levels provide protection against OA in males, with moderate physical activity levels showing a more significant protective effect.

N6-methyladenosine enhances the expression of TGF-ß-SMAD signaling family to inhibit cell growth and promote cell metastasis.

TGF-ß-SMAD signaling pathway plays an important role in the progression of various cancers. However, posttranscriptional regulation such as N6-methyladenosine (m6A) of TGF-ß-SMAD signaling axis remains incompletely understood. Here, we reveal that insulin like growth factor 2 mRNA binding protein 2 (IGF2BP2) is low expression as well as associated with poor prognosis in clear cell renal cell carcinoma (ccRCC) patients and inhibits proliferation as well as promotes metastasis of ccRCC cells. Mechanistically, IGF2BP2 systematically regulates TGF-ß-SMAD signaling family, including TGF-ß1/2, TGF-ßR1/2 and SMAD2/3/4, through mediating their mRNA stability in an m6A-dependent manner. Furthermore, the functional effects of IGF2BP2 on ccRCC cells is mediated by TGF-ß-SMAD signaling downstream effector SMAD4, which is identified three m6A sites in 5'UTR and CDS. Our study establishes IGF2BP2-TGF-ß-SMAD axis as a new regulatory effector in ccRCC, providing new insights for developing novel therapeutic strategies.

A splicing isoform of PD-1 promotes tumor progression as a potential immune checkpoint.

The immune checkpoint receptor, programmed cell death 1 (PD-1, encoded by PDCD1), mediates the immune escape of cancer, but whether PD-1 splicing isoforms contribute to this process is still unclear. Here, we identify an alternative splicing isoform of human PD-1, which carries a 28-base pairs extension retained from 5' region of intron 2 (PD-1^28), is expressed in peripheral T cells and tumor infiltrating lymphocytes. PD-1^28 expression is induced on T cells upon activation and is regulated by an RNA binding protein, TAF15. Functionally, PD-1^28 inhibits T cell proliferation, cytokine production, and tumor cell killing in vitro. In vivo, T cell-specific exogenous expression of PD-1^28 promotes tumor growth in both a syngeneic mouse tumor model and humanized NOG mice inoculated with human lung cancer cells. Our study thus demonstrates that PD-1^28 functions as an immune checkpoint, and may contribute to resistance to immune checkpoint blockade therapy.

DRAIC mediates hnRNPA2B1 stability and m6A-modified IGF1R instability to inhibit tumor progression.

Type 1 insulin-like growth factor receptor (IGF1R) plays an important role in cancer, however, posttranscriptional regulation such as N6-methyladenosine (m6A) of IGF1R remains unclear. Here, we reveal a role for a lncRNA Downregulated RNA in Cancer (DRAIC) suppress tumor growth and metastasis in clear cell Renal Carcinoma (ccRCC). Mechanistically, DRAIC physically interacts with heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1) and enhances its protein stability by blocking E3 ligase F-box protein 11 (FBXO11)-mediated ubiquitination and proteasome-dependent degradation. Subsequently, hnRNPA2B1 destabilizes m6A modified-IGF1R, leading to inhibition of ccRCC progression. Moreover, four m6A modification sites are identified to be responsible for the mRNA degradation of IGF1R. Collectively, our findings reveal that DRAIC/hnRNPA2B1 axis regulates IGF1R mRNA stability in an m6A-dependent manner and highlights an important mechanism of IGF1R fate. These findings shed light on DRAIC/hnRNPA2B1/FBXO11/IGF1R axis as potential therapeutic targets in ccRCC and build a link of molecular fate between m6A-modified RNA and ubiquitin-modified protein.

HUNK inhibits epithelial-mesenchymal transition of CRC via direct phosphorylation of GEF-H1 and activating RhoA/LIMK-1/CFL-1.

Epithelial-mesenchymal transition (EMT) is associated with the invasive and metastatic phenotypes in colorectal cancer (CRC). However, the mechanisms underlying EMT in CRC are not completely understood. In this study, we find that HUNK inhibits EMT and metastasis of CRC cells via its substrate GEF-H1 in a kinase-dependent manner. Mechanistically, HUNK directly phosphorylates GEF-H1 at serine 645 (S645) site, which activates RhoA and consequently leads to a cascade of phosphorylation of LIMK-1/CFL-1, thereby stabilizing F-actin and inhibiting EMT. Clinically, the levels of both HUNK expression and phosphorylation S645 of GEH-H1 are not only downregulated in CRC tissues with metastasis compared with that without metastasis, but also positively correlated among these tissues. Our findings highlight the importance of HUNK kinase direct phosphorylation of GEF-H1 in regulation of EMT and metastasis of CRC.

Evaluation of Biological Effects and Transcriptome Changes Induced by LED-Based Narrow-Band UVB Phototherapy.

Ultraviolet (UV), particularly UVB, is widely used in the treatment of skin diseases including psoriasis, atopic dermatitis, vitiligo, mycosis fungoides and pruritus. Recently, there has been a trend of replacing broad-band UVB (BB-UVB) units with narrow-band UVB (NB-UVB), as studies have demonstrated that NB-UVB is more efficacious in the treatment of psoriasis. The purpose of this study is to evaluate the biological effects and transcriptome changes induced by light-emitting diode-based NB-UVB (NB-UVB LED) phototherapy. Cell viability and the cell migration ability were significantly decreased posttreatment, as well as apoptosis and ROS levels were remarkably increased. NB-UVB-induced S phase arrest was observed 12 h postirradiation. Bioinformatics analysis of transcriptome sequencing data revealed that NB-UVB LED irradiation induced dose-depended changes in multiple key signaling pathways, such as PI3K and cytoskeletal-related pathways. The depolymerization of cytoskeleton induced by NB-UVB was observed 24 h posttreatment. In addition, the expression levels of cytoskeleton-related proteins FN1, ITGB4, ITGA1, RAC2 and DOCK1 decreased significantly 12 h after irradiation. Our results indicated that NB-UVB LED may serve as a novel option for the development of NB-UVB phototherapy devices.

Tumor Cell-Intrinsic BTLA Receptor Inhibits the Proliferation of Tumor Cells via ERK1/2.

B and T lymphocyte attenuator (BTLA) is an immune checkpoint molecule that mediates the escape of tumor cells from immunosurveillance. Consequently, BTLA and its ligand herpesvirus entry mediator (HVEM) are potentially immunotherapeutic targets. However, the potential effects of BTLA on tumor cells remain incompletely unknown. Here, we show that BTLA is expressed across a broad range of tumor cells. The depletion of BTLA or HVEM promotes cell proliferation and colony formation, which is reversed by the overexpression of BTLA in BTLA knockout cells. In contrast, overexpression of BTLA or HVEM inhibits tumor cell proliferation and colony formation. Furthermore, the proliferation of a subpopulation with high BTLA was also significantly slower than that of the low BTLA subpopulation. Mechanistically, the coordination of BTLA and HVEM inhibits its major downstream extracellular regulated protein kinase (ERK1/2) signaling pathway, thus preventing tumor cell growth. This study demonstrates that tumor cell-intrinsic BTLA/HVEM is a potential tumor suppressor and is likely to have a potential antagonist for immunotherapy, thus representing a potential biomarker for the optimal cancer immunotherapeutic treatment.

DMDRMR-Mediated Regulation of m6A-Modified CDK4 by m6A Reader IGF2BP3 Drives ccRCC Progression.

Aberrant N 6-methyladenosine (m6A) modification has emerged as a driver of tumor initiation and progression, yet how long noncoding RNAs (lncRNA) are involved in the regulation of m6A remains unknown. Here we utilize data from 12 cancer types from The Cancer Genome Atlas to comprehensively map lncRNAs that are potentially deregulated by DNA methylation. A novel DNA methylation-deregulated and RNA m6A reader-cooperating lncRNA (DMDRMR) facilitated tumor growth and metastasis in clear cell renal cell carcinoma (ccRCC). Mechanistically, DMDRMR bound insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) to stabilize target genes, including the cell-cycle kinase CDK4 and three extracellular matrix components (COL6A1, LAMA5, and FN1), by specifically enhancing IGF2BP3 activity on them in an m6A-dependent manner. Consequently, DMDRMR and IGF2BP3 enhanced the G1-S transition, thus promoting cell proliferation in ccRCC. In patients with ccRCC, high coexpression of DMDRMR and IGF2BP3 was associated with poor outcomes. Our findings reveal that DMDRMR cooperates with IGF2BP3 to regulate target genes in an m6A-dependent manner and may represent a potential diagnostic, prognostic, and therapeutic target in ccRCC. SIGNIFICANCE: This study demonstrates that the lncRNA DMDRMR acts as a cofactor for IGF2BP3 to stabilize target genes in an m6A-dependent manner, thus exerting essential oncogenic roles in ccRCC.

Dependencies of Surface Condensation on the Wettability and Nanostructure Size Differences.

When changing surface wettability and nanostructure size, condensation behavior displays distinct features. In this work, we investigated evaporation on a flat hydrophilic surface and condensation on both hydrophilic and hydrophobic nanostructured surfaces at the nanoscale using molecular dynamics simulations. The simulation results on hydrophilic surfaces indicated that larger groove widths and heights produced more liquid argon atoms, a quicker temperature response, and slower potential energy decline. These three characteristics closely relate to condensation areas or rates, which are determined by groove width and height. For condensation heat transfer, when the groove width was small, the change of groove height had little effect, while change of groove height caused a significant variation in the heat flux with a large groove width. When the cold wall was hydrophobic, the groove height became a significant impact factor, which caused no vapor atoms to condense in the groove with a larger height. The potential energy decreased with the increase of the groove height, which demonstrates a completely opposing trend when compared with hydrophilic surfaces.

Nitrogen-doped polymer nanofibers decorated with Co nanoparticles for uniform lithium nucleation/growth in lithium metal batteries.

Lithium (Li) metal is deemed to be the most promising anode for new-generation lithium batteries due to its high specific capacity (3860 mA h g-1) and low redox potential (-3.04 V vs. SHE). However, Li dendritic formation during battery cycling results not only in poor performance, such as low coulombic efficiency and short cycling, but also serious safety risks such as fire and explosion. In this paper, we propose a novel interlayer with a 3D network which is rich in N-containing functional groups and Co nanoparticles to guide uniform Li nucleation/growth and thus relieve the Li dendritic formation. As a result, the as-designed composite delivers an ultra-long lifespan of 1500 h with a small and stable voltage profile of 38.1 mV for symmetric cells. The composite electrode also exhibits a prominent electrochemical performance in full cells with LiFePO4 as the cathode for lithium ion batteries (LIBs). Our findings provide strong support for inducing the uniform nucleation of Li by introducing lithiophilic sites, which is important for inhibiting Li dendritic formation.

Downregulation of miR-218-5p promotes invasion of oral squamous cell carcinoma cells via activation of CD44-ROCK signaling.

The invasion front of oral squamous cell carcinoma (OSCC) harbors the most aggressive cells of the tumor and is critical for cancer invasion and metastasis. MicroRNAs (miRNAs) play important roles in OSCC progression. In this study, we modelled the OSCC invasion front on a microfluidic chip, and investigated differences in miRNA profiles between cells in the invasion front and those in the tumor mass by small RNA sequencing. We found that miR-218-5p was downregulated in invasion front cells and negatively regulates OSCC invasiveness by targeting the CD44-ROCK pathway. Thus, miR-218-5p may serve as a useful therapeutic target for OSCC. Moreover, invasion front cell isolation based-on microfluidic technology provided a useful strategy for cancer invasion study.