Abstract
Pioneer transcription factors (TFs) like SOX2 are vital for stemness and cancer through enhancing gene expression within transcriptional condensates formed with coactivators, RNAs and ...mediators on super-enhancers (SEs). Despite their importance, how these factors work together for transcriptional condensation and activation remains unclear. SOX2, a pioneer TF found in SEs of pluripotent and cancer stem cells, initiates SE-mediated transcription by binding to nucleosomes, though the mechanism isn’t fully understood. To address SOX2’s role in SEs, we identified mSE078 as a model SOX2-enriched SE and p300 as a coactivator through bioinformatic analysis. In vitro and cell assays showed SOX2 forms condensates with p300 and SOX2-binding motifs in mSE078. We further proved that SOX2 condensation is highly correlated with mSE078’s enhancer activity in cells. Moreover, we successfully demonstrated that p300 not only elevated transcriptional activity but also triggered chromatin acetylation via its direct interaction with SOX2 within these transcriptional condensates. Finally, our validation of SOX2-enriched SEs showcased their contribution to target gene expression in both stem cells and cancer cells. In its entirety, this study imparts valuable mechanistic insights into the collaborative interplay of SOX2 and its coactivator p300, shedding light on the regulation of transcriptional condensation and activation within SOX2-enriched SEs.
Graphical Abstract
Graphical Abstract
In veterinary medicine, magnetic resonance imaging (MRI) is widely utilized for brain imaging. But the complex structures of brain tissues can give rise to artifacts such as partial volume averaging ...in conventional sequences. To address this issue, several studies about double inversion recovery (DIR) sequences have been conducted in human medicine. However, published clinical studies about brain MRI using DIR sequences in dogs are currently lacking. The purpose of this study was to evaluate the magnetic resonance features of single-slab 3D DIR sequences in the normal canine brain.
Five healthy Beagle dogs were examined and the following pulse sequences were acquired for each: (1) spin-echo T2-weighted (T2W), (2) fluid attenuated inversion recovery (FLAIR), (3) gray matter (GM) selective, and (4) white matter (WM) selective single-slab 3D DIR sequence. For qualitative analysis, the distinction between gray and white matter of the cerebral cortex, presence and severity of the image artifacts were assessed for each pulse sequence. In addition, reconstructed images of single-slab 3D DIR sequences were qualitatively evaluated. For quantitative analysis, contrast ratios (CRs), signal-to-noise ratios (SNRs), and contrast-to-noise ratios (CNRs) of the GM, WM and cerebrospinal fluid (CSF) were measured for each pulse sequence.
GM selective 3D DIR was superior to T2W and FLAIR in delineating the boundaries between GM and WM in the overall brain area. Whereas WM selective 3D DIR provided better gray-white matter distinction of the cerebral cortex than T2W and FLAIR at the level of the medulla oblongata, where T2W and FLAIR images exhibited severe partial volume averaging artifacts. In general, the 3D DIR images demonstrated fewer artifacts compared to other sequences, and the reconstructed sagittal and dorsal images of these sequences maintained same spatial resolution as the original transverse images without any image degradation. Both gray and white matter selective 3D DIR sequences effectively suppressed unwanted signals, thereby providing high contrast between gray and white matter. Findings from this study could serve as a foundation for further studies on DIR sequences for the evaluation of brain diseases in dogs.
The metabolic prohormone pro-opiomelanocortin (POMC) is generally translocated into the endoplasmic reticulum (ER) for entry into the secretory pathway. Patients with mutations within the signal ...peptide (SP) of POMC or its adjoining segment develop metabolic disorders. However, the existence, metabolic fate, and functional outcomes of cytosol-retained POMC remain unclear. Here, we show that SP-uncleaved POMC is produced in the cytosol of POMC neuronal cells, thus inducing ER stress and ferroptotic cell death. Mechanistically, the cytosol-retained POMC sequesters the chaperone Hspa5 and subsequently accelerates degradation of the glutathione peroxidase Gpx4, a core regulator of ferroptosis, via the chaperone-mediated autophagy. We also show that the Marchf6 E3 ubiquitin ligase mediates the degradation of cytosol-retained POMC, thereby preventing ER stress and ferroptosis. Furthermore, POMC-Cre-mediated Marchf6-deficient mice exhibit hyperphagia, reduced energy expenditure, and weight gain. These findings suggest that Marchf6 is a critical regulator of ER stress, ferroptosis, and metabolic homeostasis in POMC neurons.
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•Marchf6 Ub ligase targets signal peptide-bearing POMC for proteasomal degradation•Cytosol-retained POMC accelerates Gpx4 degradation by chaperon-mediated autophagy•Marchf6 suppresses ER stress and ferroptosis through cytosolic POMC degradation•Marchf6 deficiency causes hyperphagia and weight gain in mice
Mun et al. show that the ER transmembrane Marchf6 E3 ubiquitin ligase promotes the proteasomal degradation of signal-peptide-containing POMC at the cytosolic face, thus preventing ER stress and ferroptosis. Marchf6 deficiency causes cytosol-retained POMC to induce Gpx4 degradation via the chaperon-mediated autophagy, leading to hyperphagia and weight gain in mice.
Shoulder joint disease is a common cause of forelimb lameness in dogs. To diagnose this condition, shoulder magnetic resonance arthrography (MRA) is performed, which involves the injection of ...contrast agents into the shoulder joint space under ultrasound (US)-guidance. The objective of this study was to compare the craniolateral and caudolateral approaches for shoulder MRA using US-guided injection techniques, and investigate their clinical feasibility in dogs. Forty shoulder joints from 10 adult beagles were studied in two repetitions. The craniolateral (n = 20) and caudolateral (n = 20) injection techniques were applied randomly under US-guidance. The shoulder MRA was conducted immediately after the contrast agents was injected. The procedure time (scan and injection time), number of attempts, joint distension and degree of extraarticular extravasation were recorded and compared between the two groups. The results showed that the caudolateral approach had significantly more contrast agents extravasation compared to the craniolateral approach (p < 0.05). However, there were no significant differences between the two groups in terms of procedure time (scan time p = 0.80, injection time p = 0.74), number of attempts (p = 0.70) and joint distension (p = 0.23). The craniolateral approach of US-guided contrast injection techniques for shoulder MRA minimizes damage to the juxta-articular structures and reduces extraarticular extravasation, resulting in good-quality images. This study demonstrates the feasibility and advantages of the craniolateral approach under US-guidance for shoulder MRA in dogs.
Adult T-cell Leukemia/Lymphoma (ATLL) is a rare aggressive T-cell malignancy caused by human T-cell leukemia virus type 1 (HTLV-1) infection. However, little is known about the underlying activated ...molecular pathways at the single cell level. Moreover, the intercellular communications between the tumor microenvironment (TME) and tumor cells in this malignancy are currently unknown. Difficulties in harvesting fresh tissue in a clinical setting have hampered our deeper understanding of this malignancy. Herein, we examined ATLL using archived fresh frozen tissue after biopsy using single-cell RNA sequencing (scRNA-seq) with T-cell receptor (TCR) clonal analysis. Highly clonal tumor cells showed multiple activating pathways, suggesting dynamic evolution of the malignancy. By dissecting diverse cell types comprising the TME, we identified a novel subset of cancer-associated fibroblast, which showed enriched epidermal growth factor receptor (EGFR)-related transcripts including early growth response 1 and 2 (EGR1 and EGR2). Cancer associated fibroblasts (CAFs) of ATLL play an important role for CD4 T-cell proliferation
FGF7-FGF1 and PDGFA-PDGFRA/B signaling, and CAFs, particularly EGR-enriched, are also associated with CD8 and NKT expansion by EGFR. These findings suggest a potential targeted therapeutic pathway to better treat this neoplasm.
The C‐type lectin‐like domain of CLEC14a (CLEC14a‐C‐type lectin‐like domain CTLD) is a key domain that mediates endothelial cell–cell contacts in angiogenesis. However, the role of CLEC14a‐CTLD in ...pathological angiogenesis has not yet been clearly elucidated. In this study, through complementarity‐determining region grafting, consecutive deglycosylation, and functional isolation, we generated a novel anti‐angiogenic human monoclonal antibody that specifically targets CLEC14a‐CTLD and that shows improved stability and homogeneity relative to the parental antibody. We found that this antibody directly inhibits CLEC14a‐CTLD‐mediated endothelial cell–cell contact and simultaneously downregulates expression of CLEC14a on the surface of endothelial cells. Using various in vitro and in vivo functional assays, we demonstrated that this antibody effectively suppresses vascular endothelial growth factor (VEGF)‐dependent angiogenesis and tumor angiogenesis of SNU182 human hepatocellular carcinoma, CFPAC‐1 human pancreatic cancer, and U87 human glioma cells. Furthermore, we also found that this antibody significantly inhibits tumor angiogenesis of HCT116 and bevacizumab‐adapted HCT116 human colorectal cancer cells. These findings suggest that antibody targeting of CLEC14a‐CTLD has the potential to suppress VEGF‐dependent angiogenesis and tumor angiogenesis and that CLEC14a‐CTLD may be a novel anti‐angiogenic target for VEGF‐dependent angiogenesis and tumor angiogenesis.
An optimized antibody, deglyco C1 IgG, that targets CLEC14a‐CTLD may be an effective strategy for suppressing CLEC14a‐mediated VEGF‐dependent and tumor angiogenesis, which is closely associated with pathological angiogenesis.
The vertebral heart scale (VHS) is the most common method used for the objective evaluation of heart size, however, VHS cannot evaluate the entire heart area. This retrospective study aimed to ...evaluate the development and feasibility of vertebral heart area ratio (VHAR = heart area/the fourth thoracic vertebra T4 body area) as a new cardiac measurement method. A total of 125 dogs considered without clinical heart abnormalities in the pre-anesthetic examination, measurements of the VHS and VHAR using thoracic radiography were compared with computed tomography measurements of the vertebral cardiac volume ratio (VCVR = cardiac volume/T4 body volume) and investigate whether the VHAR values differed between observers. The mean cardiac and T4 body volumes were 116.99 ± 108.07 cm3 and 0.92 ± 0.91 cm3, respectively. The mean values of observers 1 and 2 were 9.9 ± 0.7 v (VHS), 42.64 ± 27.94 cm2 (heart area), and 1.37 ± 0.96 cm2 (T4 body area). Intraclass coefficients were the highest for the heart area, followed by the T4 body area and VHS. The VHAR showed a moderate correlation with VHS in observers 1 (r=0.671) and 2 (r=0.633). The VCVR showed a more positive correlation with VHAR (r=0.573) than with VHS (r=0.426). These results indicated that VHAR could be used as a complement to VHS for heart size measurement, and the high degree of observer agreement for the measurements indicated the measurement reproducibility of VHAR.
Currently, Korea is undergoing significant local extreme rainfall, which contributes to more than 80% of flood disasters. Additionally, there is an increasing occurrence of such extreme rainfall in ...small stream basins, accounting for over 60% of flood disasters. Consequently, it becomes imperative to forecast runoff and water levels in advance to effectively mitigate flood disasters in small streams. The Flood Early Warning Framework (FEWF) presents one solution to reduce flood disasters by enabling the forecast of discharge and water levels during flood events. However, the application of FEWF in existing research is challenging due to the short flood travel time characteristic of small streams. This research proposes a methodology for constructing FEWF tailored to small streams using the nomograph and rating curve method. To evaluate the effectiveness of FEWF, a 6-year dataset from the Closed-circuit television-based Automatic Discharge Measurement Technique (CADMT) was utilized. The results indicate that FEWF successfully forecasts discharge and depth during flood events. By leveraging CADMT technology and real-time data, the development of precise and dependable FEWFs becomes possible. This advancement holds the potential to mitigate the consequences of extreme rainfall events and minimize flood-related casualties in small stream basins.
Shear wave elastography (SWE) induces lateral shear wave through acoustic pulses of the transducer and evaluates tissue stiffness quantitatively. This study was performed to evaluate feasibility and ...reproducibility of two-dimensional shear wave elastography (2D SWE) for evaluation of tissue stiffness and to examine technical factors that affect shear wave speed (SWS) measurements in adult dogs. Nine healthy, 2 year-old, adult beagles with the median weight of 9.8 kg were included. In this prospective, experimental, exploratory study, 2D SWE (Aplio 600) from the liver, spleen, kidneys, pancreas, prostate, lymph nodes (submandibular, retropharyngeal, axillary, medial iliac, and inguinal), submandibular salivary gland, and thyroid was performed in anesthetized beagles. Color map was drawn and SWS of each SWE were measured as Young’s modulus (kPa) and shear wave velocity (m/s). The effect of measuring site, scan approach, depth, and anesthesia on SWE was assessed in abdominal organs by two observers independently. A total of 27 SWE examinations were performed in 12 organs by each observer. All SWS measurements were preformed successfully; however, SWE in the renal medulla could not be successfully conducted, and it was excluded from further analysis. Interobserver agreement of SWE was moderate to excellent in all organs, except for the left liver lobe at 10–15 mm depth with the intercostal scan. In the liver, there was no significant effect of the measuring site and scan approach on SWE. SWS of the liver and spleen tended to be higher with increasing the depth, but no significant difference. However, anesthesia significantly increased tissue stiffness in the spleen compared to awake dog regardless of the depth (
P
< 0.05). There was a significant difference in SWS according to the measuring site in the kidneys and pancreas (
P
< 0.001). 2D SWE was feasible and highly reproducible for the estimation of tissue stiffness in dogs. Measuring site and anesthesia are sources of variability affecting SWE in abdominal organs. Therefore, these factors should be considered during SWS measurement in 2D SWE. This study provides basic data for further studies on 2D SWE on pathological conditions that may increase tissue stiffness in dogs.
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