Pathogenic Potential of Hic1-Expressing Cardiac Stromal Progenitors

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CD73 on B16F10 melanoma cells in CD73-deficient mice promotes tumor growth,angiogenesis, neovascularization,macrophage infiltration and metastasis

Ecto-5′-nucleotidase (CD73), an enzyme providing interstitial adenosine, mediates diverse physiological and pathological responses. In tumor progression, it has primarily an immunosuppressive role but is also thought to regulate neovascularization. However, the latter role is still in debate. When B16F10 melanoma was subcutaneously injected into CD73 knockout mice, changes in the tumor vasculature were not always observed. However, we demonstrated earlier that the growth and vascularization of B16F10 melanoma in CD73 knockout mice depend on the low presence of CD73 on tumor cells. To further analyze the role of CD73 on tumor growth and vascularization, we compared the changes in B16F10 melanoma subcutaneously injected into right flank of wild-type mice, CD73 knockout mice lacking host CD73 only, and CD73 knockout mice with tumor cell CD73 either inhibited with AOPCP (adenosine α,β-methylene 5′-diphosphate) or permanently knocked down through genetic modification. We report here that both inhibition and knockdown of tumor CD73 further inhibited tumor growth compared to host CD73 knockout alone. MAP-kinase signaling pathway activation also decreased more strongly in the stable knockdown. There was a significant reduction in the angiogenic activation of blood microvessels as observed by decreased anti-VEGFR2 staining. Stable CD73 knockdown also reduced endothelial cell proliferation as measured by anti-CD105 staining. However, only chemical inhibition with AOPCP significantly augmented the reduction in intratumoral microvessel density induced by host CD73 knockout. Such reduction was not observed when tumor CD73 was knocked down due to the much slower tumor growth and decreased oxygen demand as indicated by the low expression of Bad, a hypoxia marker. Decreased CD73 activity also led to the decreased expression of angiogenic factors, including VEGF and bFGF that was only partially reversed by hypoxia in tumors treated with AOPCP. Both inhibition and knockdown of tumor CD73 significantly decreased tumor macrophage infiltration and induced microenvironment changes, thereby influencing MI or MII macrophage polarization. Additionally, tumor cell CD73 is important in metastasis formation through adenosine-independent attachment to endothelium. We conclude that even low tumor cell CD73 expression has an undeniable role in melanoma progression, including the regulation of many aspects of angiogenesis. CD73 is thus a viable target in anti-angiogenic melanoma therapy.     

Circadian clock genes promote glioma progression by affecting tumour immune infiltration and tumour cell proliferation

ObjectivesCircadian rhythm controls complicated physiological activities in organisms. Circadian clock genes have been related to tumour progression, but its role in glioma is unknown. Therefore, we explored the relationship between dysregulated circadian clock genes and glioma progression.Materials and MethodsSamples were divided into different groups based on circadian clock gene expression in training dataset (n = 672) and we verified the results in other four validating datasets (n = 1570). The GO and GSEA enrichment analysis were conducted to explore potential mechanism of how circadian clock genes affected glioma progression. The single‐cell RNA‐Seq analysis was conducted to verified previous results. The immune landscape was evaluated by the ssGSEA and CIBERSORT algorithm. Cell proliferation and viability were confirmed by the CCK8 assay, colony‐forming assay and flow cytometry.ResultsThe cluster and risk model based on circadian clock gene expression can predict survival outcome. Samples were scoring by the least absolute shrinkage and selection operator regression analysis, and high scoring tumour was associated with worse survival outcome. Samples in high‐risk group manifested higher activation of immune pathway and cell cycle. Tumour immune landscape suggested high‐risk tumour infiltrated more immunocytes and more sensitivity to immunotherapy. Interfering TIMELESS expression affected circadian clock gene expression, inhibited tumour cell proliferation and arrested cell cycle at the G0/G1 phase.ConclusionsDysregulated circadian clock gene expression can affect glioma progression by affecting tumour immune landscape and cell cycle. The risk model can predict glioma survival outcome, and this model can also be applied to pan‐cancer.     

Depletion of CUL4B in macrophages ameliorates diabetic kidney disease via miR-194-5p/ITGA9 axis

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CD8+ T cells are increased in the subventricular zone with physiological and pathological aging

Age‐related cognitive decline and neurodegenerative diseases are associated with less functional neurogenic niches. It has been recently shown that aged subventricular zone (SVZ) suffers an infiltration of T cells, which affects neural stem cell activity in mice. Whether this occurs in human neurogenic niches or to which extent T‐cell infiltration is also taking place in neurodegenerative diseases remains unknown. In this work, we studied the presence of T cells in both human neurogenic niches in young and old individuals. There was a significant increase in the number of CD3⁺ and CD8⁺ T cells in the SVZ of elderly individuals, which was not detected in the dentate gyrus. Moreover, we also found CD3⁺ and CD8⁺ T cells in the SVZ of individuals with neurodegenerative diseases. However, T‐cell count was similar when compared non‐neuropathological elderly with disease diagnosed patients. Our study reveals the infiltration of T cells in old human brains, particularly in the SVZ under non‐pathological conditions and also in neurodegenerative contexts.     

Infectious clones of cotton leafroll dwarf virus strains used for genotype evaluation in cotton

Cotton blue disease (CBD) and atypical-CBD are the most important viral diseases of cotton plants in the southern region of South America. Common and atypical strains of cotton leafroll dwarf virus (CLRDV and CLRDV-at, respectively) are thought to be causative agents of CBD and atypical-CBD, respectively. Inoculation of test plants via aphid vectors is difficult, as is determining strains via molecular diagnosis; accordingly, it is difficult for breeders to evaluate the effects of blue disease-associated virus infections in cotton lineages. In the present study, we attempted to circumvent these difficulties by producing six full-length cDNA infectious clones from CLRDV and CLRDV-at strains using the Gibson Assembly protocol. For inoculation of the infectious clones, a vacuum chamber-mediated agroinfiltration protocol was adapted and applied. Using this protocol, 90%–100% of cotton plants became infected with the clones, which was not possible via syringe-based agroinfiltration. A genotyping protocol based on RT-qPCR targeting a specific region of the virus P0 protein was also developed, allowing rapid differentiation of CLRDV and CLRDV-at. Applying this protocol to 68 field samples revealed that CLRDV-at was dominant (50%) over CLRDV (5.8%) in single virus infections. These preliminary results imply that CLRDV-at might occupy the ecological niche of CLRDV in the cotton fields of Brazil.