Many tumors become addicted to autophagy for survival, suggesting inhibition of autophagy as a potential broadly applicable cancer therapy. ULK1/Atg1 is the only serine/threonine kinase in the core ...autophagy pathway and thus represents an excellent drug target. Despite recent advances in the understanding of ULK1 activation by nutrient deprivation, how ULK1 promotes autophagy remains poorly understood. Here, we screened degenerate peptide libraries to deduce the optimal ULK1 substrate motif and discovered 15 phosphorylation sites in core autophagy proteins that were verified as in vivo ULK1 targets. We utilized these ULK1 substrates to perform a cell-based screen to identify and characterize a potent ULK1 small molecule inhibitor. The compound SBI-0206965 is a highly selective ULK1 kinase inhibitor in vitro and suppressed ULK1-mediated phosphorylation events in cells, regulating autophagy and cell survival. SBI-0206965 greatly synergized with mechanistic target of rapamycin (mTOR) inhibitors to kill tumor cells, providing a strong rationale for their combined use in the clinic.
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•ULK1 phosphorylates multiple autophagy components, including VPS34 on Ser249•SBI-0206965 is a highly selective ULK1 kinase inhibitor that blocks autophagy•SBI-0206965 combined with starvation or mTOR inhibition leads to ULK1 degradation•SBI-0206965 synergizes with mTOR inhibition to induce cell death
ULK1 is a serine/threonine kinase that initiates autophagy in response to nutrient deprivation. Egan et al. define ULK1’s consensus phosphorylation motif, demonstrate that ULK1 phosphorylates several autophagy components, and develop a ULK1 small molecule inhibitor (SBI-0206965). SBI-0206965 synergizes with mTOR inhibition to enhance apoptosis in tumor cells, suggesting therapeutic opportunities.
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•Multilayer assembly of electrospun/electrosprayed PVDF-based nanofibers and beads was designed and fabricated.•The flexible PPy/PVDF electrodes were fabricated by chemical vapour ...phase polymerization.•The CsPbBr3@PVDF beads were introduced into NFMs to effectively deliver external force.•The 3D multilayer NFMs exhibited enhanced piezoelectricity and high sensitivity.
Flexible high-performance piezoelectric nanogenerators (PNG) that can convert mechanical energy into electricity are crucial for the development of self-powered wearable electronic devices. In this work, a 3D multilayer assembly of flexible organic polypyrrole (PPy) electrodes, electrospun poly(vinylidene fluoride) (PVDF) nanofibers and electrosprayed CsPbBr3@PVDF beads was proposed for a high-performance PNG, wherein two layers of CsPbBr3@PVDF beads were inserted between the top/bottom PPy electrodes and PVDF nanofibers layer, respectively. The PPy electrodes were fabricated via vapour phase polymerization of pyrrole monomers on the surface of the electrospun PVDF nanofibers with FeCl3 acted as an oxidant. As for the active piezoelectric layers, both the PVDF nanofibers and CsPbBr3@PVDF beads possessed high fractions of β crystalline phase, i.e., 83.5% and 94%, respectively, which helps to provide favorable polarization response. Besides, the introduction of the globular CsPbBr3@PVDF particles also facilitated the stress excitation to the piezoelectric layers, thus leading to enhanced output performance and high sensitivity. Piezoelectric results showed that the 3D multilayer NFMs based PNG exhibited a high open-circuit voltage of 10.3 V and a short-circuit current density of 1.29 μA/cm2, and could detect a weak pressure as low as 7.4 Pa. Furthermore, the PNG based fabrics also demonstrated excellent ability to harvest energy from the temperature fluctuations through pyroelectric effect. The structuration method of embedding globular particles into electrospun PVDF NFMs may open up the feasibility of fabricating high-output PNG.
The intestinal microbiota has been implicated in insulin resistance, although evidence regarding causality in humans is scarce. We therefore studied the effect of lean donor (allogenic) versus own ...(autologous) fecal microbiota transplantation (FMT) to male recipients with the metabolic syndrome. Whereas we did not observe metabolic changes at 18 weeks after FMT, insulin sensitivity at 6 weeks after allogenic FMT was significantly improved, accompanied by altered microbiota composition. We also observed changes in plasma metabolites such as γ-aminobutyric acid and show that metabolic response upon allogenic FMT (defined as improved insulin sensitivity 6 weeks after FMT) is dependent on decreased fecal microbial diversity at baseline. In conclusion, the beneficial effects of lean donor FMT on glucose metabolism are associated with changes in intestinal microbiota and plasma metabolites and can be predicted based on baseline fecal microbiota composition.
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•Lean donor FMT in obese metabolic syndrome patients improves insulin sensitivity•Beneficial effects of lean donor FMT are transient•Improvement in insulin sensitivity is linked to changes in plasma metabolites•Response to lean donor FMT is driven by baseline fecal microbiota composition
Kootte et al. show that fecal microbiota transplantation from lean donors to obese patients with metabolic syndrome improves insulin sensitivity, a transient effect associated with changes in microbiota composition and fasting plasma metabolites. Baseline fecal microbiota composition in recipients predicts the response to lean donor fecal microbiota transplantation.
Neoadjuvant therapy has been increasingly used to treat patients with potentially resectable pancreatic ductal adenocarcinoma (PDAC). Although the College of American Pathologists (CAP) grading ...scheme for tumor response in posttherapy specimens has been used, its clinical significance has not been validated. Previously, we proposed a 3-tier histologic tumor regression grading (HTRG) scheme (HTRG 0, no viable tumor; HTRG 1, <5% viable tumor cells; HTRG 2, ≥5% viable tumor cells) and showed that the 3-tier HTRG scheme correlated with prognosis. In this study, we sought to validate our proposed HTRG scheme in a new cohort of 167 consecutive PDAC patients who completed neoadjuvant therapy and pancreaticoduodenectomy. We found that patients with HTRG 0 or 1 were associated with a lower frequency of lymph node metastasis (P=0.004) and recurrence (P=0.01), lower ypT (P<0.001) and AJCC stage (P<0.001), longer disease-free survival (DFS, P=0.004) and overall survival (OS, P=0.02) than those with HTRG 2. However, there was no difference in either DFS or OS between the groups with CAP grade 2 and those with CAP grade 3 (P>0.05). In multivariate analysis, HTRG grade 0 or 1 was an independent prognostic factor for better DFS (P=0.03), but not OS. Therefore we validated the proposed HTRG scheme from our previous study. The proposed HTRG scheme is simple and easy to apply in practice by pathologists and might be used as a successful surrogate for longer DFS in patients with potentially resectable PDAC who completed neoadjuvant therapy and surgery.
Production of the cells that ultimately populate the thymus to generate α/β T cells has been controversial, and their molecular drivers remain undefined. Here, we report that specific deletion of ...bone-producing osteocalcin (Ocn)-expressing cells in vivo markedly reduces T-competent progenitors and thymus-homing receptor expression among bone marrow hematopoietic cells. Decreased intrathymic T cell precursors and decreased generation of mature T cells occurred despite normal thymic function. The Notch ligand DLL4 is abundantly expressed on bone marrow Ocn(+) cells, and selective depletion of DLL4 from these cells recapitulated the thymopoietic abnormality. These data indicate that specific mesenchymal cells in bone marrow provide key molecular drivers enforcing thymus-seeding progenitor generation and thereby directly link skeletal biology to the production of T cell-based adaptive immunity.
Sheep (
) and goats (
) have, for more than a millennia, been a source of fibres for human use, be it for use in clothing and furnishings, for insulation, for decorative and ceremonial purposes, or ...for combinations thereof. While use of these natural fibres has in some respects been superseded by the use of synthetic and plant-based fibres, increased accounting for the carbon and water footprint of these fibres is creating a re-emergence of interest in fibres derived from sheep and goats. The keratin-associated proteins (KAPs) are structural components of wool and hair fibres, where they form a matrix that cross-links with the keratin intermediate filaments (KIFs), the other main structural component of the fibres. Since the first report of a complete KAP protein sequence in the late 1960s, considerable effort has been made to identify the KAP proteins and their genes in mammals, and to ascertain how these genes and proteins control fibre growth and characteristics. This effort is ongoing, with more and more being understood about the structure and function of the genes. This review consolidates that knowledge and suggests future directions for research to further our understanding.
Molecular etiologies of heart failure, an emerging cardiovascular epidemic affecting 4.7 million Americans and costing 17.8 billion health-care dollars annually, remain poorly understood. Here we ...report that an inherited human dilated cardiomyopathy with refractory congestive heart failure is caused by a dominant$Arg \rightarrow Cys$missense mutation at residue 9 (R9C) in phospholamban (PLN), a transmembrane phosphoprotein that inhibits the cardiac sarcoplasmic reticular$Ca^2+-adenosine$triphosphatase (SERCA2a) pump. Transgenic$PLN^{R9C}$mice recapitulated human heart failure with premature death. Cellular and biochemical studies revealed that, unlike wild-type PLN,$PLN^{R9C}$did not directly inhibit SERCA2a. Rather,$PLN^{R9C}$trapped protein kinase A (PKA), which blocked PKA-mediated phosphorylation of wild-type PLN and in turn delayed decay of calcium transients in myocytes. These results indicate that myocellular calcium dysregulation can initiate human heart failure-a finding that may lead to therapeutic opportunities.
Yttrium-90 (Y90)-radioembolisation (RE) significantly regresses locally advanced hepatocellular carcinoma and delays disease progression. The current study is designed to deeply interrogate the ...immunological impact of Y90-RE, which elicits a sustained therapeutic response.
Time-of-flight mass cytometry and next-generation sequencing (NGS) were used to analyse the immune landscapes of tumour-infiltrating leucocytes (TILs), tumour tissues and peripheral blood mononuclear cells (PBMCs) at different time points before and after Y90-RE.
TILs isolated after Y90-RE exhibited signs of local immune activation: higher expression of granzyme B (GB) and infiltration of CD8
T cells, CD56
NK cells and CD8
CD56
NKT cells. NGS confirmed the upregulation of genes involved in innate and adaptive immune activation in Y90-RE-treated tumours. Chemotactic pathways involving CCL5 and CXCL16 correlated with the recruitment of activated GB
CD8
T cells to the Y90-RE-treated tumours. When comparing PBMCs before and after Y90-RE, we observed an increase in tumour necrosis factor-α on both the CD8
and CD4
T cells as well as an increase in percentage of antigen-presenting cells after Y90-RE, implying a systemic immune activation. Interestingly, a high percentage of PD-1
/Tim-3
CD8
T cells coexpressing the homing receptors CCR5 and CXCR6 denoted Y90-RE responders. A prediction model was also built to identify sustained responders to Y90-RE based on the immune profiles from pretreatment PBMCs.
High-dimensional analysis of tumour and systemic immune landscapes identified local and systemic immune activation that corresponded to the sustained response to Y90-RE. Potential biomarkers associated with a positive clinical response were identified and a prediction model was built to identify sustained responders prior to treatment.
Ti-Fe-Sn-Nb alloys comprised of single β-Ti phase were developed showing excellent mechanical strength and superior plasticity. The alloys exhibited increased yield strength as a function of Fe ...content, where the Ti80Fe14Sn3Nb3 alloy presented the highest yield stress (1.88GPa) comparable to that of Ti-based nanocrystalline alloys. Moreover, a significant strain-hardening (520MPa) was achieved along with the plastic deformation. The excellent mechanical strength was enhanced by the supersaturated β-Ti and the high density of lattice defects that restrict the dislocation motion.
CircRNA is a specific type of non-coding RNA that has been shown to have an important role in mammary gland (MG) activity, but no study of MG circRNA activity in sheep so far. In this study, the ...expression profile of circRNAs was investigated using RNA-Seq in MG parenchyma at peak lactation from Small-Tailed Han sheep and Gansu Alpine Merino sheep with phenotypic differences in milk yield and components. A total of 4, 906 circRNAs were found and 33 of these were differentially expressed between breeds. GO and KEGG results showed that the parental genes of differentially expressed circRNAs were mainly enriched in heterocyclic compound binding, kinase activity, adherens junction, the TGF-β signaling pathway, and the MAPK signaling pathway. This study provides an overview of circRNA expression in the ovine MG and the interaction between some key circRNAs and their target miRNAs. It improves our knowledge of the role of circRNA in sheep milk synthesis.
•The expression profile of circRNAs was investigated using RNA-Seq in ovine mammary gland of peak lactation.•A total of 4, 906 circRNAs were found and 33 of these were differentially expressed between breeds.•Parental genes of differentially expressed circRNAs were mainly enriched in some signaling pathways related to lactation.•Some target miRNAs of circRNAs were associated with mammary gland development and lactation.
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