Human fetal cerebellar cell atlas informs medulloblastoma origin and oncogenesis.

Medulloblastoma (MB) is the most common malignant childhood brain tumour1,2, yet the origin of the most aggressive subgroup-3 form remains elusive, impeding development of effective targeted treatments. Previous analyses of mouse cerebella3-5 have not fully defined the compositional heterogeneity of MBs. Here we undertook single-cell profiling of freshly isolated human fetal cerebella to establish a reference map delineating hierarchical cellular states in MBs. We identified a unique transitional cerebellar progenitor connecting neural stem cells to neuronal lineages in developing fetal cerebella. Intersectional analysis revealed that the transitional progenitors were enriched in aggressive MB subgroups, including group 3 and metastatic tumours. Single-cell multi-omics revealed underlying regulatory networks in the transitional progenitor populations, including transcriptional determinants HNRNPH1 and SOX11, which are correlated with clinical prognosis in group 3 MBs. Genomic and Hi-C profiling identified de novo long-range chromatin loops juxtaposing HNRNPH1/SOX11-targeted super-enhancers to cis-regulatory elements of MYC, an oncogenic driver for group 3 MBs. Targeting the transitional progenitor regulators inhibited MYC expression and MYC-driven group 3 MB growth. Our integrated single-cell atlases of human fetal cerebella and MBs show potential cell populations predisposed to transformation and regulatory circuitries underlying tumour cell states and oncogenesis, highlighting hitherto unrecognized transitional progenitor intermediates predictive of disease prognosis and potential therapeutic vulnerabilities.

Co-Reactive Ligand In Situ Engineered Gold Nanoclusters with Ultra-Bright Near-Infrared Electrochemiluminescence for Ultrasensitive and Label-Free Detection of Carboxylesterase Activity.

Ultrasensitive and accurate monitoring of carboxylesterase (CE) activity is extremely crucial for the early diagnosis of hepatocellular carcinoma (HCC), which is still a considerable challenge. Herein, using a co-reactive ligand engineering strategy, ultra-bright near-infrared (λmax = 830 nm) and self-enhanced electrochemiluminescence (ECL) Au nanoclusters (NCs) were in situ prepared with 2-(diethylamino) ethanethiol (DEAET) as a co-reactive ligand. Remarkably, the co-reactive ligand not only acts as a stabilizer like traditional ligands but also plays a crucial role as a co-reactant to ensure a confinement effect to shorten the charge transfer distance and increase the local concentration, significantly improving the collision efficiency between the electrogenerated free radicals. Consequently, the DEAET Au NCs exhibited a record and stable anodal ECL without the addition of an exogenous co-reactant, dramatically superior to classical Au NCs and Ru(bpy)32+ with a certain amount of the co-reactant. As a proof of concept, a convenient and label-free CE biosensor was innovatively constructed using 1-naphthyl acetate as a selective substrate, achieving ultrasensitive detection for CE activity with a low limit of detection of 9.1 × 10-7 U/L. Therefore, this work not only paves a co-reactive ligand engineering strategy for in situ preparation of high-efficiency metal NCs but also provides an ultrasensitive and convenient platform for the early diagnosis of HCC.

Correlation between hepatic oxidative damage and clinical severity and mitochondrial gene sequencing results in biliary atresia.

AIM: To assess the level of hepatic oxidative damage and its correlation with clinical severity in biliary atresia (BA), and to understand BA mitochondrial gene sequencing. METHODS: Forty-eight BA patients and 28 control subjects (20 hepatoblastoma and 8 cholestasis patients) were enrolled. Hepatic oxidative damage was assessed by the expression of oxidation and antioxidant genes, and the correlation between oxidative damage and BA incidence, liver inflammation, and fibrosis was evaluated. Moreover, 8-hydroxyguanine (8-OHdG), mitochondrial DNA (mtDNA) copy number, and mitochondrial gene sequences were determined to evaluate oxidative mtDNA damage in BA. RESULTS: The expression of oxidation gene cytochrome b-245 beta chain (CYBB) in BA was significantly increased and patients with a higher CYBB expression had the higher risk of BA incidence, liver inflammation, and cirrhosis. However, the expression of antioxidant genes was significantly decreased, and glutathione S-transferase alpha 1 (GSTA1) negatively correlated with BA incidence and cirrhosis. When GSTA1 mRNA expression was <0.5487, the sensitivity was 80.85% and the specificity was 80% for BA diagnosis. Moreover, 8-OHdG was increased, whereas mtDNA copy number was significantly decreased in BA. Using mitochondrial gene sequencing, 10 mutation sites were identified, and one family showed a maternal inheritance in genetic loci 15 326. CONCLUSIONS: In BA, oxidative damage positively correlated with BA incidence, liver inflammation, and cirrhosis. GSTA1 could be a novel diagnostic indicator. Genetic loci 15 326 could be a maternal genetic mutation site. Taken together, antioxidation therapy after Kasai surgery might have great potential in relieving liver inflammation and fibrosis in BA patients.

The HLF/IL-6/STAT3 feedforward circuit drives hepatic stellate cell activation to promote liver fibrosis.

BACKGROUND AND AIMS: Liver fibrosis is a wound-healing response that disrupts the liver architecture and function by replacing functional parenchyma with scar tissue. Recent progress has advanced our knowledge of this scarring process, but the detailed mechanism of liver fibrosis is far from clear. METHODS: The fibrotic specimens of patients and HLF (hepatic leukemia factor)PB/PB mice were used to assess the expression and role of HLF in liver fibrosis. Primary murine hepatic stellate cells (HSCs) and human HSC line Lx2 were used to investigate the impact of HLF on HSC activation and the underlying mechanism. RESULTS: Expression of HLF was detected in fibrotic livers of patients, but it was absent in the livers of healthy individuals. Intriguingly, HLF expression was confined to activated HSCs rather than other cell types in the liver. The loss of HLF impaired primary HSC activation and attenuated liver fibrosis in HLFPB/PB mice. Consistently, ectopic HLF expression significantly facilitated the activation of human HSCs. Mechanistic studies revealed that upregulated HLF transcriptionally enhanced interleukin 6 (IL-6) expression and intensified signal transducer and activator of transcription 3 (STAT3) phosphorylation, thus promoting HSC activation. Coincidentally, IL-6/STAT3 signalling in turn activated HLF expression in HSCs, thus completing a feedforward regulatory circuit in HSC activation. Moreover, correlation between HLF expression and alpha-smooth muscle actin, IL-6 and p-STAT3 levels was observed in patient fibrotic livers, supporting the role of HLF/IL-6/STAT3 cascade in liver fibrosis. CONCLUSIONS: In aggregate, we delineate a paradigm of HLF/IL-6/STAT3 regulatory circuit in liver fibrosis and propose that HLF is a novel biomarker for activated HSCs and a potential target for antifibrotic therapy.

Shp2 promotes liver cancer stem cell expansion by augmenting ß-catenin signaling and predicts chemotherapeutic response of patients.

Src-homology 2 domain-containing phosphatase 2 (Shp2) has been reported to play an important role in the maintenance and self-renewal of embryonic and adult stem cells, but its role in cancer stem cells (CSCs) remains obscure. Herein, we observed high expression of Shp2 in both chemoresistant hepatocellular carcinomas (HCCs) and recurrent HCCs from patients. A remarkable increase of Shp2 was detected in sorted epithelial cell adhesion molecule-positive or cluster of differentiation 133-positive liver CSCs and in CSC-enriched hepatoma spheroids from patients. Up-regulated Shp2 facilitated liver CSC expansion by promoting the dedifferentiation of hepatoma cells and enhancing the self-renewal of liver CSCs. Mechanistically, Shp2 dephosphorylated cell division cycle 73 in the cytosol of hepatoma cells, and the dephosphorylated cell division cycle 73 bound ß-catenin and facilitated the nuclear translocation of ß-catenin, which promoted the dedifferentiation of hepatoma cells. Shp2 increased ß-catenin accumulation by inhibiting glycogen synthase kinase 3ß-mediated ß-catenin degradation in liver CSCs, thereby enhancing the self-renewal of liver CSCs. Blockage of ß-catenin abolished the discrepancy in liver CSC proportion and the self-renewal capacity between Shp2-depleted hepatoma cells and control cells, which further confirmed that ß-catenin is required in Shp2-promoted liver CSC expansion. More importantly, HCC patients with low Shp2 levels benefited from transcatheter arterial chemoembolization or sorafenib treatment, but patients with high Shp2 expression did not, indicating the significance of Shp2 in personalized HCC therapy. CONCLUSION: Shp2 could promote HCC cell dedifferentiation and liver CSC expansion by amplifying ß-catenin signaling and may be useful in predicting patient response to chemotherapeutics. (Hepatology 2017;65:1566-1580).

Chronic inflammation-elicited liver progenitor cell conversion to liver cancer stem cell with clinical significance.

The substantial heterogeneity and hierarchical organization in liver cancer support the theory of liver cancer stem cells (LCSCs). However, the relationship between chronic hepatic inflammation and LCSC generation remains obscure. Here, we observed a close correlation between aggravated inflammation and liver progenitor cell (LPC) propagation in the cirrhotic liver of rats exposed to diethylnitrosamine. LPCs isolated from the rat cirrhotic liver initiated subcutaneous liver cancers in nonobese diabetic/severe combined immunodeficient mice, suggesting the malignant transformation of LPCs toward LCSCs. Interestingly, depletion of Kupffer cells in vivo attenuated the LCSC properties of transformed LPCs and suppressed cytokeratin 19/Oval cell 6-positive tumor occurrence. Conversely, LPCs cocultured with macrophages exhibited enhanced LCSC properties. We further demonstrated that macrophage-secreted tumor necrosis factor-α triggered chromosomal instability in LPCs through the deregulation of ubiquitin D and checkpoint kinase 2 and enhanced the self-renewal of LPCs through the tumor necrosis factor receptor 1/Src/signal transducer and activator of transcription 3 pathway, which synergistically contributed to the conversion of LPCs to LCSCs. Clinical investigation revealed that cytokeratin 19/Oval cell 6-positive liver cancer patients displayed a worse prognosis and exhibited superior response to sorafenib treatment. CONCLUSION: Our results not only clarify the cellular and molecular mechanisms underlying the inflammation-mediated LCSC generation but also provide a molecular classification for the individualized treatment of liver cancer. (Hepatology 2017;66:1934-1951).

Long non-coding RNA DILC regulates liver cancer stem cells via IL-6/STAT3 axis.

BACKGROUND & AIMS: Emerging evidence has demonstrated the aberrant expression of long non-coding RNAs (lncRNAs) in various malignancies including HCC. However, the knowledge of cancer stem cell-related lncRNAs remains limited. METHODS: lnc-DILC (lncRNA downregulated in liver cancer stem cells (LCSCs)) was identified by microarray and validated by real-time PCR. The role of lnc-DILC in LCSCs was assessed both in vitro and in vivo. Pull down assay and oligoribonucleotides or oligodeoxynucleotides treatment were conducted to evaluate the interaction between lnc-DILC and interleukin-6 (IL-6) promoter. RESULTS: Depletion of lnc-DILC markedly enhanced LCSC expansion and facilitated HCC initiation and progression, whereas ectopic expression of lnc-DILC dramatically inhibited LCSC expansion. Mechanistically, lnc-DILC inhibited the autocrine IL-6/STAT3 signaling. The putative binding locus of lnc-DILC within IL-6 promoter was confirmed by pull down assay. Consistently, the oligoribonucleotide mimics and an oligodeoxynucleotide decoy of lnc-DILC abrogated the effects on IL-6 transcription, STAT3 activation and LCSC expansion triggered by lnc-DILC depletion and lnc-DILC overexpression. Moreover, our data suggested that lnc-DILC mediated the crosstalk between TNF-α/NF-κB signaling and IL-6/STAT3 cascade. Clinical investigation demonstrated the reduction of lnc-DILC in patient HCCs, and suggested the correlation between lnc-DILC levels and IL-6, EpCAM or CD24 expression. Decreased lnc-DILC expression in HCCs predicts early recurrence and short survival of patients, highlighting its prognostic value. CONCLUSIONS: lnc-DILC mediates the crosstalk between TNF-α/NF-κB signaling and autocrine IL-6/STAT3 cascade and connects hepatic inflammation with LCSC expansion, suggesting that lnc-DILC could be not only a potential prognostic biomarker, but also a possible therapeutic target against LCSCs.

Single-cell multi-omics analysis of lineage development and spatial organization in the human fetal cerebellum.

Human cerebellum encompasses numerous neurons, exhibiting a distinct developmental paradigm from cerebrum. Here we conducted scRNA-seq, scATAC-seq and spatial transcriptomic analyses of fetal samples from gestational week (GW) 13 to 18 to explore the emergence of cellular diversity and developmental programs in the developing human cerebellum. We identified transitory granule cell progenitors that are conserved across species. Special patterns in both granule cells and Purkinje cells were dissected multidimensionally. Species-specific gene expression patterns of cerebellar lobes were characterized and we found that PARM1 exhibited inconsistent distribution in human and mouse granule cells. A novel cluster of potential neuroepithelium at the rhombic lip was identified. We also resolved various subtypes of Purkinje cells and unipolar brush cells and revealed gene regulatory networks controlling their diversification. Therefore, our study offers a valuable multi-omics landscape of human fetal cerebellum and advances our understanding of development and spatial organization of human cerebellum.

MicroRNA-4287 alleviates inflammatory response via targeting RIPK1 in osteoarthritis.

Studies have confirmed the regulatory effects of microRNAs (miRNAs) in osteoarthritis (OA) progression. MiR-4287 has been identified by a previous study as a downregulated miRNA in chondrocytes treated with IL-1ß and TNF-α. However, the function of the underlying mechanism of miR-4287 in OA is elusive. IL-1ß-treated chondrocytes were used as OA cell models. RNA expression was accessed using RT-qPCR. Cell Counting Kit-8 (CCK-8) assay was used to determine the chondrocytes' viability and proliferation. The protein levels of inflammation factors (IL-8, IL-6, and TNF-α), matrix metalloproteinases (MMP 1, MMP3, MMP13), and chondrogenic genes (COL2A1, SOX9, and Aggrecan) were detected using western blot analysis. Luciferase reporter assays were performed for interaction exploration. HE staining and Safranin O/Fast Green staining was used to access the pathological changes in OA mouse tissues and cartilage degeneration in OA mouse. MiR-4287 was downregulated in chondrocytes treated with IL-1ß and OA mouse models. MiR-4287 overexpression promoted the viability, and proliferation and attenuated the inflammation response and destruction of cartilage in IL-1ß-stimulated chondrocytes. Receptor-interacting protein kinase 1 (RIPK1) was a target gene of miR-4287 in chondrocytes. MiR-4287 negatively regulated RIPK1 expression. RIPK1 overexpression was revealed to reverse the miR-4287-mediated effects on proliferation and inflammatory response in IL-1ß-stimulated chondrocytes. Moreover, miR-4287 was demonstrated to inhibit the pathological changes, cartilage degeneration and inflammation response in OA mice models. In conclusion, miR-4287 is a critical molecule in OA development, which attenuates inflammatory response in vivo and in vitro by targeting RIPK1.

Self-Powered Multifunction Ionic Skins Based on Gradient Polyelectrolyte Hydrogels.

Human skin is the largest organ, and it can transform multiple external stimuli into the biopotential signals by virtue of ions as information carriers. Ionic skins (i-skins) that can mimic human skin have been extensively explored; however, the limited sensing capacities as well as the need of an extra power supply significantly restrict their broad applications. Herein, we develop self-powered humanlike i-skins based on gradient polyelectrolyte membranes (GPMs) that can directly and accurately perceive multiple stimuli. Prepared by a hydrogel-assisted reaction-diffusion method, the GPMs exhibit gradient-distributed charged groups across polymer networks, enabling one to generate a thickness-dependent and thermoresponsive self-induced potential in a hydrated situation and in a humidity-sensitive self-induced potential in a dehydrated/dried situation, respectively. Consequently, the GPM-based i-skins can precisely detect pressure, temperature, and humidity in a self-powered manner. The coupling of mechano-electric and thermo-electric effects inherent in GPMs provides a general strategy for developing innovative self-powered ion-based perception systems.

A multi-omic approach reveals utility of CD45 expression in prognosis and novel target discovery.

CD45, the leukocyte common antigen, is expressed on almost all cells of the immunological and hematological systems. CD45 expression is related to a variety of diseases, including leukemia and lymphoma. In this study, we analyzed the expression level of CD45 across cancers and evaluated the relationship between its expression and patient prognosis. We further integrated methylation data to explore the differences in CD45 across cancers from a multi-omics perspective. We also analyzed the relationship between CD45 expression and levels of immune cell infiltrates and immune modifiers. Our results revealed the distinct expression characteristics and prognostic value of CD45 across multiple tumors. In addition, we screened drug targets based on the immune index defined by CD45 expression and identified that GPR84 affected the proliferation of tumor cells and was associated with the inflammation caused by immunotherapy. In summary, our findings provide a comprehensive understanding of the role of CD45 in oncogenesis and its prognostic significance across cancers.

Transcriptional Networks Identify BRPF1 as a Potential Drug Target Based on Inflammatory Signature in Primary Lower-Grade Gliomas.

Gliomas are the most common tumors of the central nervous system and are classified into grades I-IV based on their histological characteristics. Lower-grade gliomas (LGG) can be divided into grade II diffuse low-grade gliomas and grade III moderate gliomas and have a relatively good prognosis. However, LGG often develops into high-grade glioma within a few years. This study aimed to construct and identify the prognostic value of an inflammatory signature and discover potential drug targets for primary LGG. We first screened differentially expressed genes in primary LGG (TCGA) compared with normal brain tissue (GTEx) that overlapped with inflammation-related genes from MSigDB. After survival analysis, nine genes were selected to construct an inflammatory signature. LGG patients with a high inflammatory signature score had a poor prognosis, and the inflammatory signature was a strong independent prognostic factor in both the training cohort (TCGA) and validation cohort (CGGA). Compared with the low-inflammatory signature group, differentially expressed genes in the high-inflammatory signature group were mainly enriched in immune-related signaling pathways, which is consistent with the distribution of immune cells in the high- and low-inflammatory signature groups. Integrating driver genes, upregulated genes and drug targets data, bromodomain and PHD finger-containing protein 1 (BRPF1) was selected as a potential drug target. Inhibition of BRPF1 function or knockdown of BRPF1 expression attenuated glioma cell proliferation and colony formation.

Biomechanical evaluation of axial-loading simulated experiment in wrist fractures: a finite element analysis.

OBJECTIVE: To assess the biomechanical properties that influence wrist fracture, so as to provide the theoretical basis for simulation experiments to aid the optimal design of wrist protectors. METHODS: Six cadaveric wrists were included as experimental specimens. Wrist specimens wearing wrist protectors formed the experimental group and unprotected wrist specimens formed the control group. The wrist specimens were axially loaded under physiological loads and the stress magnitude and distribution of the experimental and control groups were obtained. A three-dimensional wrist finite element model of a healthy volunteer was developed to verify the rationality and effectiveness of the cadaveric wrist models. RESULTS: Under normal physiological loads, the stress on the radioulnar palmar unit was high and manifested in the form of pressure, while the stress on the radioulnar dorsal unit was lower and manifested in the form of tension. The stresses on the radial distal palmar, ulnar distal palmar, radial distal dorsal, ulnar distal dorsal, radial proximal palmar and ulnar proximal palmar units in the experimental group were less than those in the control group. CONCLUSION: Under physiological loads, wearing a wrist protector can reduce the stress on the radioulnar distal palmar, radioulnar proximal palmar and radioulnar distal dorsal units, while having no obvious effect on the radioulnar proximal dorsal units.

Fucoidan exerts protective effects against diabetic nephropathy related to spontaneous diabetes through the NF-κB signaling pathway in vivo and in vitro.

Fucoidan, an extract of the seaweed, Fucus vesiculosus, has been widely investigated for its antioxidant effects. However, to date and to the best of our knowledge, pathological studies on the effects of fucoidan against diabetic nephropathy (DN) related to spontaneous diabetes have not been carried out. DN is one of the most serious microvascular complications of diabetes. Therefore, in the present study, the effects of fucoidan against DN related to spontaneous diabetes were investigated in vitro and in vivo. Goto-Kakizaki (GK) rats were allowed free access to standard rat food with or without fucoidan for 13 weeks, and Wistar rats were used as controls. Fucoidan did not show any cytotoxicity on glomerular mesangial cells (GMCs) which were separated from rat kidneys. Fasting blood glucose levels were measured using a blood glucose meter, blood urea nitrogen (BUN) and serum creatinine (Cr) levels were measured using an automatic biochemistry analyzer and urine protein levels were measured using an ELISA kit. Collagen Ⅳ levels in the renal cortex were measured using an ELISA kit, and the expression levels of transforming growth factor-ß1 (TGF-ß1) and fibronectin (FN) in the renal cortex and GMCs, and nuclear factor-κB (NF-κB) in GMCs were determined by western blot analysis. Fasting blood glucose, BUN, serum Cr, urine protein and collagen Ⅳ levels, and the expression of TGF-ß1 and FN, as well as NF-κB p65 nuclear translocation all significantly increased in the GK rats compared with the control Wistar rats. The increase in the fasting blood glucose, BUN, serum Cr, urine protein and collagen Ⅳ levels in the renal cortex was reversed in the GK rats which were orally administered fucoidan. The oral administration of fucoidan also decreased the expression of TGF-ß1 and FN in the renal cortex and GMCs, as well as the nuclear translocation of NF-κB p65 in the GMCs. Taken together, the data from our in vitro and in vivo experiments indicate that fucoidan attenuates hyperglycemia and prevents or impedes the development of DN related to spontaneous diabetes by attenuating the activation of the NF-κB signaling pathway.