Patients with metastatic cancer experience a severe loss of skeletal muscle mass and function known as cachexia. Cachexia is associated with poor prognosis and accelerated death in patients with ...cancer, yet its underlying mechanisms remain poorly understood. Here, we identify the metal-ion transporter ZRT- and IRT-like protein 14 (ZIP14) as a critical mediator of cancer-induced cachexia. ZIP14 is upregulated in cachectic muscles of mice and in patients with metastatic cancer and can be induced by TNF-α and TGF-β cytokines. Strikingly, germline ablation or muscle-specific depletion of Zip14 markedly reduces muscle atrophy in metastatic cancer models. We find that ZIP14-mediated zinc uptake in muscle progenitor cells represses the expression of MyoD and Mef2c and blocks muscle-cell differentiation. Importantly, ZIP14-mediated zinc accumulation in differentiated muscle cells induces myosin heavy chain loss. These results highlight a previously unrecognized role for altered zinc homeostasis in metastatic cancer-induced muscle wasting and implicate ZIP14 as a therapeutic target for its treatment.
Recent work has highlighted the tumor microenvironment as a central player in cancer. In particular, interactions between tumor and immune cells may help drive the development of brain tumors such as ...glioblastoma multiforme (GBM). Despite significant research into the molecular classification of glioblastoma, few studies have characterized in a comprehensive manner the immune infiltrate in situ and within different GBM subtypes.In this study, we use an unbiased, automated immunohistochemistry-based approach to determine the immune phenotype of the four GBM subtypes (classical, mesenchymal, neural and proneural) in a cohort of 98 patients. Tissue Micro Arrays (TMA) were stained for CD20 (B lymphocytes), CD5, CD3, CD4, CD8 (T lymphocytes), CD68 (microglia), and CD163 (bone marrow derived macrophages) antibodies. Using automated image analysis, the percentage of each immune population was calculated with respect to the total tumor cells. Mesenchymal GBMs displayed the highest percentage of microglia, macrophage, and lymphocyte infiltration. CD68
and CD163
cells were the most abundant cell populations in all four GBM subtypes, and a higher percentage of CD163
cells was associated with a worse prognosis. We also compared our results to the relative composition of immune cell type infiltration (using RNA-seq data) across TCGA GBM tumors and validated our results obtained with immunohistochemistry with an external cohort and a different method. The results of this study offer a comprehensive analysis of the distribution and the infiltration of the immune components across the four commonly described GBM subgroups, setting the basis for a more detailed patient classification and new insights that may be used to better apply or design immunotherapies for GBM.
A new benzimidazole-functionalized calix4arene receptor (R) was synthesized and characterized. The receptor R shows better selectivity toward trinitrotoluene (TNT) compared to the other nitro ...explosives in solution, which also retains its effectiveness for solid-phase detection. The chemical interactions of the molecule with different nitro explosive analytes were studied by fluorescence spectroscopy and by a molecular dynamics approach. The molecular dynamics studies show a 1:3 complex between R and TNT, and hence high sensitivity was imparted by fluorescence studies. The detection of explosive vapors in ambient conditions was tested by using a sensitive coating layer of R on an SU-8/CB-based piezoresistive cantilever surface. The developed device showed large sensitivity toward TNT compared to cyclotrimethylenetrinitramine (RDX) and pentaerythritol tetranitrate (PETN) in the solid state at their respective vapor pressures at room temperature. The detection sensitivity of the device was estimated to be 35 mV for TNT at ambient conditions. Moreover, the sensor does not show a response when exposed to humidity. These results demonstrate that R can be used as one of the coating materials for a cantilever for the detection of TNT using piezoresistivity measurement. R can also detect the explosives in solution with high sensitivity and selectivity by fluorescence spectroscopy.
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IJS, KILJ, NUK, PNG, UL, UM
In cystic fibrosis (CF), impaired immune cell responses, driven by the dysfunctional CF transmembrane conductance regulator (CFTR) gene, may determine the disease severity but clinical heterogeneity ...remains a major therapeutic challenge. The characterization of molecular mechanisms underlying impaired immune responses in CF may reveal novel targets with therapeutic potential. Therefore, we utilized simultaneous RNA sequencing targeted at identifying differentially expressed genes, transcripts, and miRNAs that characterize impaired immune responses triggered by CF and its phenotypes.
Peripheral blood mononuclear cells (PBMCs) extracted from a healthy donor were stimulated with plasma from CF patients (n = 9) and healthy controls (n = 3). The PBMCs were cultured (1 × 10
cells/well) for 9 h at 37 ° C in 5% CO
. After culture, total RNA was extracted from each sample and used for simultaneous total RNA and miRNA sequencing.
Analysis of expression signatures from peripheral blood mononuclear cells induced by plasma of CF patients and healthy controls identified 151 genes, 154 individual transcripts, and 41 miRNAs differentially expressed in CF compared to HC while the expression signatures of 285 genes, 241 individual transcripts, and seven miRNAs differed due to CF phenotypes. Top immune pathways influenced by CF included agranulocyte adhesion, diapedesis signaling, and IL17 signaling, while those influenced by CF phenotypes included natural killer cell signaling and PI3K signaling in B lymphocytes. Upstream regulator analysis indicated dysregulation of CCL5, NF-κB and IL1A due to CF while dysregulation of TREM1 and TP53 regulators were associated with CF phenotype. Five miRNAs showed inverse expression patterns with three target genes relevant in CF-associated impaired immune pathways while two miRNAs showed inverse expression patterns with two target genes relevant to a dysregulated immune pathway associated with CF phenotypes.
Our results indicate that miRNAs and individual transcript variants are relevant molecular targets contributing to impaired immune cell responses in CF.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Donor‐derived cell‐free DNA (dd‐cfDNA) has been evaluated as a rejection marker in organ transplantation. This study sought to assess the utility of dd‐cfDNA to diagnose graft injury in liver ...transplant recipients (LTR) and as a predictive biomarker prior to different causes of graft dysfunction. Plasma from single and multicenter LTR cohorts was analyzed for dd‐cfDNA. Phenotypes of treated biopsy‐proven acute rejection (AR, N = 57), normal function (TX, N = 94), and acute dysfunction no rejection (ADNR; N = 68) were divided into training and test sets. In the training set, dd‐cfDNA was significantly different between AR versus TX (AUC 0.95, 5.3% cutoff) and AR versus ADNR (AUC 0.71, 20.4% cutoff). Using these cutoffs in the test set, the accuracy and NPV were 87% and 100% (AR vs. TX) and 66.7% and 87.8% (AR vs. ADNR). Blood samples collected serially from LTR demonstrated incremental elevations in dd‐cfDNA prior to the onset of graft dysfunction (AR > ADNR), but not in TX. Dd‐cfDNA also decreased following treatment of rejection. In conclusion, the serial elevation of dd‐cfDNA identifies pre‐clinical graft injury in the context of normal liver function tests and is greatest in rejection. This biomarker may help detect early signs of graft injury and rejection to inform LTR management strategies.
In independent discovery and validation cohorts, this study reports the utility of donor‐derived cell‐free DNA as an early sensitive biomarker of rejection and graft injury in liver transplant recipients, supporting its potential in optimizing serial patient management.
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BFBNIB, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
-Methyladenosine (m
A) is the most abundant modification of mammalian mRNAs. RNA methylation fine tunes RNA stability and translation, altering cell fate. The fat mass- and obesity-associated protein ...(FTO) is an m
A demethylase with oncogenic properties in leukemia. Here, we show that
expression is suppressed in ovarian tumors and cancer stem cells (CSC). FTO inhibited the self-renewal of ovarian CSC and suppressed tumorigenesis
, both of which required FTO demethylase activity. Integrative RNA sequencing and m
A mapping analysis revealed significant transcriptomic changes associated with
overexpression and m
A loss involving stem cell signaling, RNA transcription, and mRNA splicing pathways. By reducing m
A levels at the 3'UTR and the mRNA stability of two phosphodiesterase genes (
and
), FTO augmented second messenger 3', 5'-cyclic adenosine monophosphate (cAMP) signaling and suppressed stemness features of ovarian cancer cells. Our results reveal a previously unappreciated tumor suppressor function of FTO in ovarian CSC mediated through inhibition of cAMP signaling. SIGNIFICANCE: A new tumor suppressor function of the RNA demethylase FTO implicates m
A RNA modifications in the regulation of cyclic AMP signaling involved in stemness and tumor initiation.
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•Biofunctionalization of DNA aptamer against cardiac marker protein Troponin-T on ZnO channel surface.•Characterization of biofunctionalized surface using XPS and AFM techniques.•A ...novel approach (KPFM) to understand the efficacy of biomolecular interaction between cardiac Troponin-T and its corresponding aptamer.•Fabrication and characterization of ZnO-TFT as a sensor device for Troponin-T detection.
In this work, we have demonstrated the use of DNA aptamer as a selective bio recognizable entity towards the qualitative detection of cardiac Troponin-T (cTnT), a clinically important cardiac marker protein. Prior to this, the ZnO thin film surface as an active semiconducting channel material was first biofunctionalized with DNA aptamer by using standard biotin-streptavidin chemistry. Biofunctionalization was further confirmed by analyzing the ZnO surface with atomic force microscope (AFM) and X- ray photoelectron spectroscopy (XPS) techniques. In order to understand the efficacy of biomolecular interaction between cardiac Troponin-T (cTnT) and its corresponding aptamer, Kelvin Probe Force Microscopy (KPFM) technique was performed to monitor the effective contact potential difference. Further, to realize the essential technological importance of this methodology as a sensor platform, a thin-film transistor (TFT) micro device was fabricated using ZnO as a channel material and characterized. These fabricated devices were further used to demonstrate the qualitative detection of cardiac Troponin-T protein by employing aptamer-mediated biofunctionalization of ZnO TFT.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Noninvasive biomarkers distinguishing early immune activation before acute rejection (AR) could more objectively inform immunosuppression management in liver transplant recipients (LTRs). We ...previously reported a genomic profile distinguishing LTR with AR versus stable graft function. This current study includes key phenotypes with other causes of graft dysfunction and uses a novel random forest approach to augment the specificity of predicting and diagnosing AR.
Gene expression results in LTRs with AR versus non-AR (combination of other causes of graft dysfunction and normal function) were analyzed from single and multicenter cohorts. A 70:30 approach (61 ARs; 162 non-ARs) was used for training and testing sets. Microarray data were normalized using a LT-specific vector.
Random forest modeling on the training set generated a 59-probe classifier distinguishing AR versus non-AR (area under the curve 0.83; accuracy 0.78, sensitivity 0.70, specificity 0.81, positive predictive value 0.54, negative predictive value NPV 0.89; F-score 0.61). Using a locked threshold, the classifier performed well on the testing set (accuracy 0.72, sensitivity 0.67, specificity 0.73, positive predictive value 0.48, NPV 0.86; F-score 0.56). Probability scores increased in samples preceding AR versus non-AR, when liver function tests were normal, and decreased following AR treatment (P < 0.001). Ingenuity pathway analysis of the genes revealed a high percentage related to immune responses and liver injury.
We have developed a blood-based biologically relevant biomarker that can be detected before AR-associated graft injury distinct from LTR never developing AR. Given its high NPV ("rule out AR"), the biomarker has the potential to inform precision-guided immunosuppression minimization in LTRs.
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