Ideal Target Research Articles

MiRNA-449c-5p regulates the JAK-STAT pathway in inhibiting cell proliferation and invasion in human breast cancer cells by targeting ERBB2.

Breast cancer is a highly prevalent disease worldwide, and early diagnosis and treatment could reduce the mortality rate of breast cancer patients. microRNAs (miRNA) have been shown to regulate the occurrences and progression of many types of cancers. Thus, it is crucial to find novel biomarkers in breast cancer. miR-449c-5p acted as a biomarker in non-small cell lung cancer, gastric carcinoma, and so forth. ERBB2 is an ideal target for breast cancer therapy. However, the molecular mechanisms between miR-449c-5p and ERBB2 in breast cancer remain poorly understood. Our study focused on the regulatory role of miR-449c-5p in breast cancer and its targeting relationship with ERBB2. The miR-449c-5p expression in breast cancer tissue and normal tissue was searched from the online database (Starbase). The clinical prognosis of miR-449c-5p and ERBB2 was predicted by using the Kaplan-Meier analysis method. The expression of miR-449c-5p mimics and inhibitors was measured by qRT-PCR. T47D cells were transfected with miR-449c-5p mimics and miR-449c-5p inhibitors. After that, CCK-8, colony formation assays and Transwell assays were used to evaluate the cell proliferation ability, migration and invasion. Whether ERBB2 was the target gene of the miR-449c-5p was predicted by Starbase and verified by dual-luciferase activity assay. In addition, protein levels and the relationship between signalling pathways were measured and validated using western blotting analysis. We confirmed that miR-449c-5p was highly expressed in breast cancer tissue, and its downregulation was linked with poor prognosis. Overexpression of miR-449c-5p inhibited the proliferation, migration and invasion of breast cancer cells. ERBB2 was a target of miR-449c-5p. The invasion, migration, and proliferation of breast cancer cells were inhibited by miR-449c-5p/ERBB2 through JAK-STAT. This study demonstrated that miR-449c-5p inhibits breast cancer cell proliferation, migration and invasion by targeting ERBB2 via JAK/STAT, which means miR-449c-5p, is a potential biomarker for breast cancer and provides a novel insight for diagnosis.

A method for fast reconstruction of closed section workpiece surface using point clouds

Parameterized reconstruction of workpiece surfaces is a critical step for advanced manufacturing applications, such as the solving of machining parameters, and the expression of workpiece in situ measurement. Generally, the point clouds model is easy to obtain and can be seen as the source model of the workpiece, and the B-spline surface is always taken as the ideal target parameterized model. Existing research focuses on reconstructing the single-valued surface, which can be projected onto a spatial plane without overlapping. However, the dual-valued surface, which overlaps at least twice, is also commonly encountered in industry, and has few investigations. Therefore, this paper proposes a fast reconstruction method based on reasonable segmentation and parameter domain mapping. In this approach, first, the dual-valued point clouds model is segmented into multiple single-valued point clouds models according to the sought edges in the optimal attitude. Then, the grid point coordinates of the single-valued point clouds model are calculated using the parametric-domain moving least squares method. Finally, the B-spline surface is generated by interpolating these grid points. The proposed algorithm is applied to single-bending closed-section workpieces such as the turbine blade to illustrate the effectiveness of this method. The contrast results with existing method demonstrate that the proposed approach can reconstruct the workpiece with multi-valued surfaces more quickly and more accurately, which is significant in advanced manufacturing.

Chasing the Break: Tracing the Full Evolution of a Black Hole X-Ray Binary Jet with Multiwavelength Spectral Modeling

Black hole (BH) X-ray binaries (XRBs) are ideal targets to study the connection between accretion inflow and jet outflow. Here we present quasi-simultaneous, multiwavelength observations of the Galactic BH system MAXI J1820+070, throughout its 2018–2019 outburst. Our data set includes coverage from the radio through X-ray bands from 17 different instruments/telescopes, and encompasses 19 epochs over a 7 month period, resulting in one of the most well-sampled multiwavelength data sets of a BH XRB outburst to date. With our data, we compile and model the broadband spectra of this source using a phenomenological model that includes emission from the jet, a companion star, and an accretion flow. This modeling allows us to track the evolution of the spectral break in the jet spectrum, a key observable that samples the jet launching region. We find that the spectral break location changes over at least ≈3 orders of magnitude in electromagnetic frequency over this period. Using these spectral break measurements, we link the full cycle of jet behavior, including the rising, quenching, and reignition, to the changing accretion flow properties as the source evolves through its different accretion states. Our analysis shows consistent jet behavior with other sources in similar phases of their outbursts, reinforcing the idea that jet quenching and recovery may be a global feature of BH XRB systems in outburst. Our results also provide valuable evidence supporting a close connection between the geometry of the inner accretion flow and the base of the jet.

Eocene monsoon climate expansion in East Asia: Evidence from orbital‐cycle driven terrestrial successions in the Jianghan Basin, Central China

ABSTRACTEast Asia experienced complex climatic changes during the Eocene. Due to the lack of a high‐precision time framework covering the Eocene, the characteristics of sedimentary and climate evolution in this period remain unclear. The Jianghan Basin (Central China) developed an Eocene continental succession, which is composed of lower red beds and upper salt‐lake deposits exhibiting clear sedimentary rhythms, making these strata an ideal target for astronomical cycle analysis. Based on the established floating astronomical timescale, the lower red beds in the basin formed during the early–middle Eocene and were dominated by eccentricity cycles. The upper salt‐lake deposits formed during the middle–late Eocene and recorded obliquity cycles. The change in facies indicates that the climate transitioned from continuously arid to less arid with periodic relative wetness, which was forced by orbital cycles. Significant obliquity signals recorded by salt rhythmites responded to Eocene global temperature changes and Tibetan region uplift, which can be important markers of a monsoon‐like climate. Comparing the sedimentary and climatic characteristics of other Eocene lake basins in East Asia, the extension of the Eocene East Asian monsoon may have initiated during ca 43.6 to 40.0 Ma. This study provides a new perspective for understanding the early evolution of the East Asian monsoon.

Identifying virulence factors using graph transformer autoencoder with ESMFold-predicted structures

With the increasing resistance of bacterial pathogens to conventional antibiotics, antivirulence strategies targeting virulence factors (VFs) have become an effective new therapy for the treatment of pathogenic bacterial infections. Therefore, the identification and prediction of VFs can provide ideal candidate targets for the implementation of antivirulence strategies in treating infections caused by pathogenic bacteria. Currently, the existing computational models predominantly rely on the amino acid sequences of virulence proteins while overlooking structural information. Here, we propose a novel graph transformer autoencoder for VF identification (GTAE-VF), which utilizes ESMFold-predicted 3D structures and converts the VF identification problem into a graph-level prediction task. In an encoder-decoder framework, GTAE-VF adaptively learns both local and global information by integrating a graph convolutional network and a transformer to implement all-pair message passing, which can better capture long-range correlations and potential relationships. Extensive experiments on an independent test dataset demonstrate that GTAE-VF achieves reliable and robust prediction accuracy with an AUC of 0.963, which is consistently better than that of other structure-based and sequence-based approaches. We believe that GTAE-VF has the potential to emerge as a valuable tool for assessing VFs and devising antivirulence strategies.

Structural and functional validation of a highly specific Smurf2 inhibitor.

Smurf1 and Smurf2 are two closely related member of the HECT (homologous to E6AP carboxy terminus) E3 ubiquitin ligase family and play important roles in the regulation of various cellular processes. Both were initially identified to regulate transforming growth factor-β and bone morphogenetic protein signaling pathways through regulating Smad protein stability and are now implicated in various pathological processes. Generally, E3 ligases, of which over 800 exist in humans, are ideal targets for inhibition as they determine substrate specificity; however, there are few inhibitors with the ability to precisely target a particular E3 ligase of interest. In this work, we explored a panel of ubiquitin variants (UbVs) that were previously identified to bind Smurf1 or Smurf2. In vitro binding and ubiquitination assays identified a highly specific Smurf2 inhibitor, UbV S2.4, which was able to inhibit ligase activity with high potency in the low nanomolar range. Orthologous cellular assays further demonstrated high specificity of UbV S2.4 toward Smurf2 and no cross-reactivity toward Smurf1. Structural analysis of UbV S2.4 in complex with Smurf2 revealed its mechanism of inhibition was through targeting the E2 binding site. In summary, we investigated several protein-based inhibitors of Smurf1 and Smurf2 and identified a highly specific Smurf2 inhibitor that disrupts the E2-E3 protein interaction interface.

An antisense oligonucleotide efficiently suppresses splicing of an alternative exon in vascular smooth muscle in vivo.

Targeting alternative exons for therapeutic gain has been achieved in a few instances and potentially could be applied more broadly. The myosin phosphatase (MP) enzyme is a critical hub upon which signals converge to regulate vessel tone. Alternative exon 24 of myosin phosphatase regulatory subunit (Mypt1 E24) is an ideal target as toggling between the two isoforms sets smooth muscle sensitivity to vasodilators such as nitric oxide (NO). This study aimed to develop a gene-based therapy to suppress splicing of Mypt1 E24 thereby switching MP enzyme to the NO-responsive isoform. CRISPR/Cas9 constructs were effective at editing of Mypt1 E24 in vitro; however, targeting of vascular smooth muscle in vivo with AAV9 was inefficient. In contrast, an octo-guanidine conjugated antisense oligonucleotide targeting the 5' splice site of Mypt1 E24 was highly efficient in vivo. It reduced the percent splicing inclusion of Mypt1 E24 from 80% to 10% in mesenteric arteries. The maximal and half-maximal effects occurred at 12.5 and 6.25 mg/kg, respectively. The effect persisted for at least 1 mo without toxicity. This highly effective splice-blocking antisense oligonucleotide could be developed as a novel therapy to reverse vascular dysfunction common to diseases such as hypertension and heart failure.NEW & NOTEWORTHY Alternative exon usage is a major driver of phenotypic diversity in all cell types including smooth muscle. However, the functional significance of most of the hundreds of thousands of alternative exons has not been defined, nor in most cases even tested. If their importance to vascular function were known these alternative exons could represent novel therapeutic targets. Here, we present injection of Vivo-morpholino splice-blocking antisense oligonucleotides as a simple, efficient, and cost-effective method for suppression of alternative exon usage in vascular smooth muscle in vivo.

Optimization of the Spot Spacings for Reducing Roughness in Laser-Induced Optical Breakdown Processes for Corneal Laser Vision Correction

The aim of this work is to implement an algorithm that simulates a simplified cutting surface based on laser-induced optical breakdown (LIOB). The algorithm includes the definition of a possible positioning of the laser pulses and calculation of the roughness for different parameter settings (including LIOB threshold, pulse energies, and spot spacings) as the difference between simulated and ideal target cut (local differences within the cut, i.e., the waviness of the simulated cut vs. the homogeneity of the ideal target cut). Furthermore, optimizations of specific variables, such as spot distance (along the pathway), track distance (between lines/tracks), and pulse energy, are performed. The simulations suggest that lower pulse energies (well above the LIOB threshold) combined with asymmetric spacings (spot-to-track distance ratio >> 1) may be effective to lower the roughness of laser cuts generated by LIOB processes. The importance of lowering pulse energies (well above the threshold) emphasizes the need for the LIOB threshold to remain low (as low as possible). Reducing roughness by decreasing spacings (thus, increasing dose for same pulse energies) may have negative implications in visual recovery (risk for overdose). In all cases, the roughness is multiple times larger (rougher) than equivalent simulations for ablative procedures.

EVALUATING THE PRE-CLINICAL EFFICACY OF VT1021 (A CLINICAL STAGE DRUG CANDIDATE) IN MULTIPLE EXPERIMENTAL MODELS OF INFLAMMATORY BOWEL DISEASE

Abstract Inflammatory bowel disease (IBD) is characterized symptomatically by chronic inflammation in the colonic tract. Despite the increased understanding of the physiological symptoms of Crohn’s Disease, and IBD in general, there have been no significant breakthroughs in understanding its etiology or improving treatment. As such, there is an urgent need for efficacious therapies that are aimed specifically at preventing or curing IBD. Bolstering the need for better therapies is the fact that patients with IBD have increased risk of developing intestinal cancers. One of the pathological traits of IBD that heightens the risk of cancer is the increased level of angiogenesis that accompanies the inflammation. As the biological processes of angiogenesis and inflammation are intricately linked, we believe increasing the levels of thrombospondin-1 (TSP-1), an anti-angiogenic and anti-inflammatory protein, can reprogram immune and inflammatory cells to inhibit angiogenesis and induce the resolution of inflammation. Vigeo has developed VT1021 which stimulates the expression of TSP-1 in inflammatory myeloid derived suppressor cells (MDSCs) systemically. VT1021 achieved proof of concept in a Phase 1/2 clinical trial with multiple cohorts and is currently in Phase 2/3 trials for oncology indications. IBD patients have increased MDSCs in both lesions and peripheral blood. MDSCs are genomically stable and not mutated in IBD patients and thus represent ideal therapeutic targets. Two dextran sulfate sodium (DSS)-induced colitis models were used to evaluate the pre-clinical efficacy of VT1021. In the preventative model, mice were treated with 4% DSS from Day 0 to Day 6 and either saline or VT1021 from Day 0 to Day 8, then sacrificed on Day 8 to collect colon tissue and plasma samples. In the rescue model, mice were treated with 4% DSS from Day 0 to Day 4, then treated with either saline, VT1021 (25, 50, 100 mg/kg) or anti-TNFa from Day 5 to Day 10. Mice were sacrificed on Day 11; colon tissue and plasma samples were collected. In both models, VT1021 treatment significantly prevented the mouse weight loss and bloody stool compared to the saline treatment. VT1021 treatment also decreased the colon inflammation by the measurement of both plasma myeloperoxidase (MPO) level and colon tissue histopathology score. Immunohistochemistry staining of colon tissue samples confirmed that VT1021 treatment stimulated TSP-1 expression in the MDSCs. Altogether, these findings show that VT1021 treatment has equivalent efficacy to anti-TNFa treatment, while significantly reducing damage to colon tissue. We believe VT1021 will show significantly greater efficacy in patients with longer term use, while avoiding the systemic deleterious side effects common to current approved therapies.

Redshifted Iron Emission and Absorption Lines in the Chandra X-Ray Spectrum of Centaurus A

Cen A hosts the closest active galactic nucleus to the Milky Way, which makes it an ideal target for investigating the dynamical processes in the vicinity of accreting supermassive black holes. In this paper, we present 14 Chandra HETGS spectra of the nucleus of Cen A that were observed throughout 2022. We compared them with each other, and contrasted them against the two previous Chandra HETGS spectra from 2001. This enabled an investigation into the spectral changes occurring on timescales of months and 21 years. All Chandra spectra could be well fitted by an absorbed power law with a strong and narrow Fe Kα line, a leaked power-law feature at low energies, and Si and S Kα lines that could not be associated with the central engine. The flux of the continuum varied by a factor of 2.74 ± 0.05 over the course of the observations, whereas the Fe line only varied by 18.8% ± 8.8%. The photon index increased over 21 years, and the hydrogen column density varied significantly within a few months as well. The Fe Kα line was found at a lower energy than expected from the Cen A redshift, amounting to an excess velocity of 326−94+84kms−1 relative to Cen A. We investigated warped accretion disks, bulk motion, and outflows as possible explanations of this shift. The spectra also featured ionized absorption lines from Fe xxv and Fe xxvi, describing a variable inflow.

Visual and whole-body quantitative analyses of 68Ga-DOTATATE PET/CT for prognosis of outcome after PRRT with 177Lu-DOTATATE.

Somatostatin receptors (SSTR) represent an ideal target for nuclear theranostics applications in neuroendocrine tumors (NET). Studies suggest that high uptake on SSTR-PET is associated with response to SSTR peptide receptor radionuclide therapy (PRRT). The purpose of this study was to evaluate the role of baseline whole-body (WB) 68Ga-DOTATATE PET/CT (SSTR-PET) quantitative parameters, and the presence of NET lesions without uptake on SSTR-PET, as outcome prognosticator in patients with NET treated with PRRT. Patients with NET who underwent at least 4 177Lu-DOTATATE PRRT cycles between 07/2016 and 03/2021 were included in this retrospective analysis if they fulfilled the following inclusion criteria: SSTR-PET within 6months of 1st PRRT cycle, follow-up CT and/or MRI performed > 6months after the 4th cycle of PRRT. The SSTR-PET analysis consisted of a visual and a quantitative analysis done independently by two board-certified physicians. The visual analysis assessed the presence of NET lesions visible on the SSTR-PET co-registered CT. The quantitative analysis consisted in contouring all SSTR-avid lesions on SSTR-PET and extracting WB quantitative parameters: SUVmean (WB-SUVmean), SUVmax of the lesion with highest uptake (H-SUVmax), and tumor volume (WB-TV). WB-SSTR-PET parameters and the presence of SSTR-PET-negative lesions were correlated to radiologic response (assessed by RECIST 1.1 criteria) and progression-free survival (PFS). Fisher's exact test, Mann-Whitney's U test and Kaplan-Meier curves with Cox-regression analysis were used for the statistical analysis. Forty patients (F/M: 21/19; 34/40 with gastro-entero-pancreatic (GEP) NET, 6/40 with non-GEP NET) were included in the analysis. The median follow-up period after the 4th PRRT cycle was 25.7months (range 15.2-59.1). Fourteen/40 (35%) patients showed radiologic response (RECIST PR). PFS event was observed in 17/40 (42.5%) patients. Thirteen/40 (32.5%) patients had SSTR-PET-negative lesions at baseline. Higher WB-SUVmean and H-SUVmax were associated with better response (p = 0.015 and 0.005, respectively). The presence of SSTR-PET-negative lesions and lower WB-SUVmean were associated with shorter PFS (p = 0.026 and 0.008, respectively). Visual and quantitative analyses of baseline SSTR-PET can yield valuable information to prognosticate outcomes after 177Lu-DOTATATE PRRT.

Lysine methyltransferase SMYD2 enhances androgen receptor signaling to modulate CRPC cell resistance to enzalutamide.

Androgen receptors (ARs) play key roles in prostate cancer (PCa) progression and castration-resistant prostate cancer (CRPC) resistance to drug therapy. SET and MYND domain containing protein 2 (SMYD2), a lysine methyltransferase, has been reported to promote tumors by transcriptionally methylating important oncogenes or tumor repressor genes. However, the role of SMYD2 in CRPC drug resistance remains unclear. In this study, we found that SMYD2 expression was significantly upregulated in PCa tissues and cell lines. High SMYD2 expression indicated poor CRPC-free survival and overall survival in patients. SMYD2 knockdown dramatically inhibited the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) potential of 22Rv1 and C4-2 cells. Conversely, ectopic overexpression of SMYD2 promoted these effects in 22Rv1 and C4-2 cells. Mechanistically, SMYD2 methylated and phosphorylated ARs to affect AR ubiquitination and proteasome degradation, which further alters the AR transcriptome in CRPC cells. Importantly, the SMYD2 inhibitor AZ505 had a synergistic therapeutic effect with enzalutamide in CRPC cells and mouse models; however, it could also re-sensitize resistant CRPC cells to enzalutamide. Our findings demonstrated that SMYD2 enhances the methylation and phosphorylation of ARs and affects AR ubiquitination and proteasome degradation to modulate CRPC cell resistance to enzalutamide, indicating that SMYD2 serves as a crucial oncogene in PCa and is an ideal therapeutic target for CRPC.

Design, Synthesis, and Biological Evaluation of 2-Substituted Aniline Pyrimidine Derivatives as Potent Dual Mer/c-Met Inhibitors.

Mer and c-Met kinases, which are commonly overexpressed in various tumors, are ideal targets for the development of antitumor drugs. This study focuses on the design, synthesis, and evaluation of several 2-substituted aniline pyrimidine derivatives as highly potent dual inhibitors of Mer and c-Met kinases for effective tumor treatment. Compound 18c emerged as a standout candidate, demonstrating robust inhibitory activity against Mer and c-Met kinases, with IC50 values of 18.5 ± 2.3 nM and 33.6 ± 4.3 nM, respectively. Additionally, compound 18c displayed good antiproliferative activities on HepG2, MDA-MB-231, and HCT116 cancer cells, along with favorable safety profiles in hERG testing. Notably, it exhibited exceptional liver microsomal stability in vitro, with a half-life of 53.1 min in human liver microsome. Compound 18c also exhibited dose-dependent cytotoxicity and hindered migration of HCT116 cancer cells, as demonstrated in apoptosis and migration assays. These findings collectively suggest that compound 18c holds promise as a dual Mer/c-Met agent for cancer treatment.

Versatile CYP98A enzymes catalyse meta-hydroxylation reveals diversity of salvianolic acids biosynthesis.

Salvianolic acids (SA), such as rosmarinic acid (RA), danshensu (DSS), and their derivative salvianolic acid B (SAB), etc. widely existed in Lamiaceae and Boraginaceae families, are of interest due to medicinal properties in the pharmaceutical industries. Hundreds of studies in past decades described that 4-coumaroyl-CoA and 4-hydroxyphenyllactic acid (4-HPL) are common substrates to biosynthesize SA with participation of rosmarinic acid synthase (RAS) and cytochrome P450 98A (CYP98A) subfamily enzymes in different plants. However, in our recent study, several acyl donors and acceptors included DSS as well as their ester-forming products all were determined in SA-rich plants, which indicated that previous recognition to SA biosynthesis is insufficient. Here, we used Salvia miltiorrhiza, a representative important medicinal plant rich in SA, to elucidate the diversity of SA biosynthesis. Various acyl donors as well as acceptors are catalysed by SmRAS to form precursors of RA and two SmCYP98A family members, SmCYP98A14 and SmCYP98A75, are responsible for different positions' meta-hydroxylation of these precursors. SmCYP98A75 preferentially catalyses C-3' hydroxylation, and SmCYP98A14 preferentially catalyses C-3 hydroxylation in RA generation. In addition, relative to C-3' hydroxylation of the acyl acceptor moiety in RA biosynthesis, SmCYP98A75 has been verified as the first enzyme that participates in DSS formation. Furthermore, SmCYP98A enzymes knockout resulted in the decrease and overexpression leaded to dramatic increase of SA accumlation. Our study provides new insights into SA biosynthesis diversity in SA-abundant species and versatility of CYP98A enzymes catalytic preference in meta-hydroxylation reactions. Moreover, CYP98A enzymes are ideal metabolic engineering targets to elevate SA content.

Incongruity between T cell receptor recognition of breast cancer hotspot mutations ESR1 Y537S and D538G following exogenous peptide loading versus endogenous antigen processing

T cell receptor engineered T cell (TCR T) therapies have shown recent efficacy against certain types of solid metastatic cancers. However, to extend TCR T therapies to treat more patients across additional cancer types, new TCRs recognizing cancer-specific antigen targets are needed. Driver mutations in AKT1, ESR1, PIK3CA, and TP53 are common in patients with metastatic breast cancer (MBC) and if immunogenic could serve as ideal tumor-specific targets for TCR T therapy to treat this disease. Through IFN-γ ELISpot screening of in vitro expanded neopeptide-stimulated T cell lines from healthy donors and MBC patients, we identified reactivity towards 11 of 13 of the mutations. To identify neopeptide-specific TCRs, we then performed single-cell RNA sequencing of one of the T cell lines following neopeptide stimulation. Here, we identified an ESR1 Y537S specific T cell clone, clonotype 16, and an ESR1 Y537S/D538G dual-specific T cell clone, clonotype 21, which were HLA-B*40:02 and HLA-C*01:02 restricted, respectively. TCR Ts expressing these TCRs recognized and killed target cells pulsed with ESR1 neopeptides with minimal activity against ESR1 WT peptide. However, these TCRs failed to recognize target cells expressing endogenous mutant ESR1. To investigate the basis of this lack of recognition we performed immunopeptidomics analysis of a mutant-overexpressing lymphoblastoid cell line and found that the ESR1 Y537S neopeptide was not endogenously processed, despite binding to HLA-B*40:02 when exogenously pulsed onto the target cell. These results indicate that stimulation of T cells that likely derive from the naïve repertoire with pulsed minimal peptides may lead to the expansion of clones that recognize non-processed peptides, and highlights the importance of using methods that selectively expand T cells with specificity for antigens that are efficiently processed and presented.

A transcriptional regulation of ERF15 contributes to ABA-mediated cold tolerance in tomato.

Cold stress is a major meteorological threat to crop growth and yield. Abscisic acid (ABA) plays important roles in plant cold tolerance by activating the expression of cold-responsive genes; however, the underlying transcriptional regulatory module remains unknown. Here, we demonstrated that the cold- and ABA-responsive transcription factor ETHYLENE RESPONSE FACTOR 15 (ERF15) positively regulates ABA-mediated cold tolerance in tomato. Exogenous ABA treatment significantly enhanced cold tolerance in wild-type tomato plants but failed to rescue erf15 mutants from cold stress. Transcriptome analysis showed that ERF15 was associated with the expression of cold-responsive transcription factors such as CBF1 and WRKY6. Further RT-qPCR assays confirmed that the ABA-induced increased in CBF1 and WRKY6 transcripts was suppressed in erf15 mutants when the plants were subjected to cold treatment. Moreover, yeast one-hybrid assays, dual-luciferase assays and electrophoretic mobility shift assays demonstrated that ERF15 activated the transcription of CBF1 and WRKY6 by binding their promoters. Silencing CBF1 or WRKY6 significantly decreased cold tolerance. Overall, our study identified the role of ERF15 in conferring ABA-mediated cold tolerance in tomato plants by activating CBF1 and WRKY6 expression. This study not only broadens our knowledge of the mechanism of ABA-mediated cold tolerance in plants but also highlights ERF15 as an ideal target gene for cold-tolerant crop breeding.

TMPRSS2 isoform 1 downregulation by G-quadruplex stabilization induces SARS-CoV-2 replication arrest

BackgroundSARS-CoV-2 infection depends on the host cell factors angiotensin-converting enzyme 2, ACE2, and the transmembrane serinprotease 2, TMPRSS2. Potential inhibitors of these proteins would be ideal targets against severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection. Our data opens the possibility that changes within TMPRSS2 can modulate the outcome during a SARS-CoV-2 infection.ResultsWe reveal that TMPRSS2 acts not only during viral entry but has also an important role during viral replication. In addition to previous functions for TMPRSS2 during viral entry, we determined by specific downregulation of distinct isoforms that only isoform 1 controls and supports viral replication. G-quadruplex (G4) stabilization by chemical compounds impacts TMPRSS2 gene expression. Here we extend and in-depth characterize these observations and identify that a specific G4 in the first exon of the TMPRSS2 isoform 1 is particular targeted by the G4 ligand and affects viral replication. Analysis of potential single nucleotide polymorphisms (SNPs) reveals that a reported SNP at this G4 in isoform 1 destroys the G4 motif and makes TMPRSS2 ineffective towards G4 treatment.ConclusionThese findings uncover a novel mechanism in which G4 stabilization impacts SARS-CoV-2 replication by changing TMPRSS2 isoform 1 gene expression.

Detail mineralogical study and geochronological framework of Bayan Obo (China) Nb mineralization recorded by in situ U-Pb dating of columbite

The Bayan Obo deposit is world-famous for its giant rare earth reserves, and also accompanied with 2.2 Mt Nb2O5 resources at an average grade of 0.13 %, renowned as the second largest Nb deposit globally. Previous studies have focused on the origin of carbonatite (igneous/sedimentary), and variation composition during evolution, as well as its super-enrichment of light rare earth elements (LREE), while the metallogenic mechanism of niobium is being overlooked despite the identification of plenty Nb-bearing minerals. In this study, we identify four types of columbite as major Nb-bearing minerals, instead of pyrochlore in the west pit of Bayan Obo. Detailed mineralogical study has revealed large primitive columbite (Clb1-I, 100–200 μm) is hosted in the coarse-grained Fe-dolomite carbonatite (FeO = 5.46–10.47 wt%), being an ideal target for future mineral beneficiation. Those columbites (Clb1-I) are featured by lowest Sr, Y, and ∑REE but enriched in iron in composition, revealing its magmatic origin. Sieve-textured columbite (Clb1-II, ∼100 μm) dispersed in the fluorination and chloritization dolomite carbonatite, with a U–Pb age of 1359 ± 31 Ma, contemporary with ages of the regional carbonatite magma intrusion and rift-related magmatism, indicating niobium being sourced from mantle-derived carbonatite. The occurrence of fluorite as inclusions in Clb1-I indicates the solubility of Nb was enhanced by fluxes, and the columbite saturation was easily achieved during the carbonatite magma evolution into ferroan carbonatite. Strong fenitization metasomatism further locally concentrated columbite either within the mineral assemblages of pyrite + phlogopite + allanite (Clb2) or in the mica-dominated fenite (Clb3), and crystallized at 346 ± 18 Ma and 284 ± 16 Ma, respectively. The latter metalogenetic epoch is accordant with late Permian granitic magmatism event (ca. 280 Ma) while the former age may attribute to its occurrences related to the early Paleozoic arc-continent collision. Differ from pristine columbite (Clb1-I and Clb1-II), the metamorphic columbite (Clb2 and Cl3, especially Clb3) are enriched in Ta (1803–4092 ppm), Y (560–2029 ppm) and ∑REE (720–6317 ppm) content, probably suggesting the highest degree of evolution. The similar metallogenesis framework between Nb and REE, combined with the coexistence of columbite and REE minerals assemblages, implies the spatial and temporal coupling of Nb and REE mineralization. This first innovative and systematic mineralogical and geochronological work directly constrains the nature of Mesoproterozoic mantle-derived carbonatite with fertile Fe and fluxes, and the Paleozoic and Permian fenitization metasomatism, are the major factors contribute to the metallogenic specialization of niobium in Bayan Obo.

Mismatch repair is not correlated with genomic alterations in glioblastoma patients

Glioma is the most common brain tumor of the glial cells with a highly infiltrative nature. A grade IV glioblastoma is the most malignant and aggressive presenting with complex mutations and a very high genomic alteration status. Glioblastoma patients have the fastest progression compared to other cancer patients and poor survival. Methylation status of MGMT, the gene encoding a DNA repair enzyme, is the only known promising biomarker. There is a need to look for new biomarkers in similar functional pathways. Two mismatch repair pathway (MMR) genes, hMSH2, and hMSH6, which function downstream of MGMT, appear as ideal targets. We hypothesized that a lower expression of hMSH2 and hMSH6 are associated with a higher fraction of genome-altered (FGA) which would influence poor survival. We conducted an in-silico study using publicly available datasets consisting of clinical and experimental data. We obtained a weak positive correlation between hMSH2 and hMSH6 with FGA. Sex specific analyses yielded a stronger significant positive correlation. However, overall the measures of correlation were too small as the component analysis indicated homology and validated the findings. Additionally, the expression of hMSH2 and hMSH6 did not affect overall survival. We conclude that in this cohort hMSH2 and hMSH6 did not indicate biological significance in causing FGA, nor were they effective biomarkers for survival as hypothesized.

Learning the Autoimmune Pathogenesis Through the Study of Aire.

One of the difficulties in studying the pathogenesis of autoimmune diseases is that the disease is multifactorial involving sex, age, MHC, environment, and some genetic factors. Because deficiency of Aire, a transcriptional regulator, is an autoimmune disease caused by a single gene abnormality, Aire is an ideal research target for approaching the enigma of autoimmunity, e.g., the mechanisms underlying Aire deficiency can be studied using genetically modified animals. Nevertheless, the exact mechanisms of the breakdown of self-tolerance due to Aire's dysfunction have not yet been fully clarified. This is due, at least in part, to the lack of information on the exact target genes controlled by Aire. State-of-the-art research infrastructures such as single-cell analysis are now in place to elucidate the essential function of Aire. The knowledge gained through the study of Aire-mediated tolerance should help our understanding of the pathogenesis of autoimmune disease in general.