Cell Differentiation Activity Research Articles

BMP Gene-Immobilization to Dental Implants Enhances Bone Regeneration.

For individuals who have experienced tooth loss, dental implants are an important treatment option for oral reconstruction. For these patients, alveolar bone augmentation and acceleration of osseointegration optimize implant stability. Traditional oral surgery often requires invasive procedures, which can result in prolonged treatment time and associated morbidity. It has been previously shown that chemical vapor deposition (CVD) polymerization of functionalized [2.2]paracyclophanes can be used to anchor gene encoding vectors onto biomaterial surfaces and local delivery of a bone morphogenetic protein (BMP)-encoding vector can increase alveolar bone volume and density in vivo. This study is the first to combine the use of CVD technology and BMP gene delivery on titanium for the promotion of bone regeneration and bone to implant contact in vivo. BMP-7 tethered to titanium surface enhances osteoblast cell differentiation and alkaline phosphatase activity in vitro and increases alveolar bone regeneration and % bone to implant contact similar to using high doses of exogenously applied BMP-7 in vivo. The use of this innovative gene delivery strategy on implant surfaces offers an alternative treatment option for targeted alveolar bone reconstruction.

Abstract 2879: Preclinical characterization of GT-00A x IL15: A novel IL-15-based immunocytokine with unique tumor targeting properties

Abstract Introduction: IL-15 is a potent pro-inflammatory cytokine that enhances the differentiation, proliferation and cytolytic activity of NK cells and T cells. Due to the huge potential of IL-15 to activate both innate and adaptive anti-tumor immunity, several IL-15-based immunocytokines are currently in clinical development. However, all of them preferentially act in the periphery and not locally within the tumor. To further increase the efficacy and safety of IL-15-based immunocytokines, we developed GT-00A x IL15, an immunocytokine targeting a tumor-associated, glycosylated epitope of MUC-1 (TA-MUC1). GT-00A x IL15 was designed to balance best tumor enrichment, efficacy and safety and thus represents a new cancer immunotherapeutic for the treatment of solid tumors with high medical need. Experimental procedures: GT-00A x IL15 was extensively studied in several in vitro assays to assess its activity and characterize its complex mechanisms of action. Several pharmacokinetic, pharmacodynamic, biodistribution and efficacy studies were performed in tumor-free or tumor-bearing mice to further analyze its pharmacological profile and efficacy in vivo. The safety of GT-00A x IL15 was assessed in a 4-weeks GLP toxicology study in rats. Results summary: We can demonstrate in vitro that targeting IL-15 to tumor cells by using GT-00A x IL15 increases the cytotoxic activity of effector cells against TA-MUC1-positive tumor cell lines compared to parental GT-00A and an untargeted IL-15 control construct. Further, GT-00A x IL15 mediates a dose-dependent infiltration of NK and T cells into MCF-7 tumor spheroids that is not observed using the controls. In a biodistribution study using radioactively labelled GT-00A x IL15, we observed significantly increased enrichment of GT-00A x IL15 in the tumor compared to the untargeted IL-15 control construct. This tumor enrichment translated into activation and expansion of tumor-infiltrating NK and CD8+ T cell effector cells and single agent efficacy in different tumor models in vivo. The safety assessment of GT-00A x IL15 in a dose-range finding study using rats as relevant species did not show any safety concerns so far. The full data package will be available early 2022 to be presented at the AACR meeting. Conclusions: Our results show that by using TA-MUC1 as a broadly expressed and highly tumor-specific antigen, we are able to direct IL-15 to the tumor. Local enrichment of GT-00A x IL15 within the tumor ultimately induced local effector cell activation and expansion. GT-00A x IL15 showed single agent efficacy in vivo with a good safety profile. These data highlight the potential of GT-00A x IL15 to increase the efficacy and safety of an IL-15-based immunocytokine by tumor targeting. GT-00A x IL15 shows great promise for the treatment of TA-MUC1-positive solid tumors either as monotherapeutic agent or as valuable combination partner. Citation Format: Patrik Kehler, Anika Jaekel, Johanna Gellert, Christoph Goletz, Timo Lischke, Anke Flechner, Antje Danielczyk, Theresa Neumann. Preclinical characterization of GT-00A x IL15: A novel IL-15-based immunocytokine with unique tumor targeting properties [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2879.

Panax notoginseng Saponins Stimulates Neurogenesis and Neurological Restoration After Microsphere-Induced Cerebral Embolism in Rats Partially Via mTOR Signaling.

P. Notoginseng Saponins (PNS), the main active component of herbal medicine Panax notoginseng, has been widely used to treat cerebrovascular diseases. It has been acknowledged that PNS exerted protection on nerve injuries induced by ischemic stroke, however, the long-term impacts of PNS on the restoration of neurological defects and neuroregeneration after stroke have not been thoroughly studied and the underlying molecular mechanism of stimulating neurogenesis is difficult to precisely clarify, much more in-depth researches are badly needed. In the present study, cerebral ischemia injury was induced by microsphere embolism (ME) in rats. After 14 days, PNS administration relieved cerebral ischemia injury as evidenced by alleviating neurological deficits and reducing hippocampal pathological damage. What’s more, PNS stimulated hippocampal neurogenesis by promoting cell proliferation, migration and differentiation activity and modulated synaptic plasticity. Increased number of BrdU/Nestin, BrdU/DCX and NeuroD1-positive cells and upregulated synapse-related GAP43, SYP, and PSD95 expression were observed in the hippocampus. We hypothesized that upregulation of brain-derived neurotrophic factor (BDNF) expression and activation of Akt/mTOR/p70S6K signaling after ME could partially underlie the neuroprotective effects of PNS against cerebral ischemia injury. Our findings offer some new viewpoints into the beneficial roles of PNS against ischemic stroke.

EBV/HHV-6A dUTPases contribute to myalgic encephalomyelitis/chronic fatigue syndrome pathophysiology by enhancing TFH cell differentiation and extrafollicular activities.

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a chronic, debilitating, multisystem illness of unknown etiology for which no cure and no diagnostic tests are available. Despite increasing evidence implicating EBV and human herpesvirus 6A (HHV-6A) as potential causative infectious agents in a subset of patients with ME/CFS, few mechanistic studies address a causal relationship. In this study we examined a large ME/CFS cohort and controls and demonstrated a significant increase in activin A and IL-21 serum levels, which correlated with seropositivity for antibodies against the EBV and HHV-6 protein deoxyuridine triphosphate nucleotidohydrolase (dUTPases) but no increase in CXCL13. These cytokines are critical for T follicular helper (TFH) cell differentiation and for the generation of high-affinity antibodies and long-lived plasma cells. Notably, ME/CFS serum was sufficient to drive TFH cell differentiation via an activin A–dependent mechanism. The lack of simultaneous CXCL13 increase with IL-21 indicates impaired TFH function in ME/CFS. In vitro studies revealed that virus dUTPases strongly induced activin A secretion while in vivo, EBV dUTPase induced the formation of splenic marginal zone B and invariant NKTFH cells. Together, our data indicate abnormal germinal center (GC) activity in participants with ME/CFS and highlight a mechanism by which EBV and HHV6 dUTPases may alter GC and extrafollicular antibody responses.

MEDB-60. Medulloblastoma with extensive nodularity mimics cerebellar development and differentiates along the granular precursor lineage

BACKGROUND: Medulloblastoma with extensive nodularity (MBEN) represents a rare type of cerebellar tumors of infancy comprising two histologically distinct components that differ in cell differentiation and mitotic activity. Whereas some children suffering from MBEN experience disease recurrence, MBEN can also spontaneously differentiate and discontinue to grow. The underlying mechanisms of this variable biological behavior may offer insight into how embryonal tumors develop. METHODS: Fresh frozen and FFPE tumor tissue from nine MBEN-patients was subjected to multi-omics characterization including bulk sequencing, microdissection followed by RNA sequencing, single nucleus RNA-sequencing using the 10X Genomics- and SMART Seq. V2-protocols and spatial transcriptomics via RNAscope. RESULTS: All cases were molecularly classified as Sonic Hedgehog (SHH)-MB, and harbored somatic mutations within the SHH-pathway. After quality control, a total of ~30.000 cells were subjected to downstream analysis. Several non-malignant cell types, such as glial cells, were identified. In accordance with previous studies, we found only sparse immune infiltration. Unsupervised clustering identified cell clusters that differed in differentiation state and represented a continuum from embryonal-like cells with SHH-upregulation over intermediate cell states, to neuronal-like, postmitotic cells. Mapping to a single nucleus sequencing atlas of cerebellar development indicated that tumor cells reflected various stages of normally developing cerebellar granular precursors. Interestingly, one cluster of malignant cells with tumor-specific copy number alterations showed both transcriptomic features of astrocytes and embryonal cells. Using spatial transcriptomics, we were able to correlate different clusters of MBEN cells with distinct histologic MBEN compartments, with astrocyte-like tumor cells being located in the internodular compartment and in close proximity to mitotically active cancer cells. CONCLUSION: MBEN is formed by a continuum of malignant cell differentiation along the granular precursor lineage, with a subset of cells developing into cells that may represent tumor astrocytes. This differentiation process is reflected in the bicompartmental structure of MBEN.

Non-coding RNAs regulating epithelial-mesenchymal transition: Research progress in liver disease.

Chronic liver injury could gradually progress to liver fibrosis, cirrhosis, and even hepatic carcinoma without effective treatment. The massive production and activation of abnormal cell differentiation is vital to the procession of liver diseases. Epithelial-mesenchymal transformation (EMT) is a biological process in which differentiated epithelial cells lose their epithelial characteristics and acquire mesenchymal cell migration capacity. Emerging evidence suggests that EMT not only occurs in the process of hepatocellular carcinogenesis, but also appears in liver cells transforming to myofibroblasts, a core event of liver disease. Non-coding RNA (ncRNA) such as microRNA (miRNA), long non-coding RNA (lncRNA) and circular RNA (circRNA) are important regulatory factors in EMT, which can regulate target gene expression by binding with RNA single-stranded. Various studies had shown that ncRNA regulation of EMT plays a key role in liver disease development, and many effective ncRNAs have been identified as promising biomarkers for the diagnosis and treatment of liver disease. In this review, we focus on the relationship between the different ncRNAs and EMT as well as the specific molecular mechanism in the liver diseases to enrich the pathological progress of liver diseases and provide reference for the treatment of liver diseases.

Absence of Toll-like receptor 5 confers survival in mice bearing ovarian tumors treated with anti-PD-L1

Abstract Ovarian cancer accounts for more deaths than any other cancer of the female reproductive system. Patients bearing ovarian tumors infiltrated with high frequencies of T cells associate with a greater survival probability. However, therapeutic strategies targeting T cells in ovarian cancers are largely ineffective. We have observed in our models of late-stage murine ovarian cancer that in the absence of Toll-Like Receptor 5 (TLR5) signaling, anti-PD-L1 therapy promotes significant survival and subsequent protection against tumor rechallenge. Ovarian tumors from rechallenged TLR5KO mice exhibited significantly higher frequency/number of both CD4 and CD8 effector memory T cell subsets and greater numbers of antigen experienced T cells. Thus, in the absence of TLR5 signaling, T cells are better able to mount/maintain a response to ovarian tumors after inhibitory receptor blockade. Furthermore, protection against tumor progression in TLR5KO mice corresponds with a significant increase in tumor infiltrating cross presenting and IL-12 producing Dendritic cells (DCs) which could account for the differences in T cell infiltrates. We hypothesize that TLR5 signaling functionally alters DCs to become more suppressive in the tumor microenvironment, impacting T cell differentiation and anti-tumor activity, thereby reducing survival during ovarian tumor progression with aPD-L1 therapy. Clinical implications are already present, as roughly 7.5% of the general population harbor a TLR5 SNP that diminishes TLR5 signaling and is associated with increased long-term survival for ovarian cancer patients. Therefore, patients who express the TLR5 SNP may immediately benefit from anti-PDL1 therapy and those without the SNP, TLR5 antagonism. Supported by NIH/NCI 1R01CA253285 UVA CIC Collaborative Grant

MyD88-mediated signaling in myo-/fibroblasts is required for control of macrophage maturation under mucosal tolerance in the gut

Abstract Introduction Colonic myo-/fibroblasts (MFs) are cells of mesenchymal origin in a healthy gut are among the key players in the mucosal tolerance that involve MF-restricted MyD88 signaling. MFs and macrophages interact in colonic tissue. However, the mechanisms of MF mediated regulation of macrophages under intestinal mucosal tolerance has emerging evidence. We hypothesized that MF-restricted MyD88 signaling regulates macrophage maturation under colonic mucosal tolerance. Methods RNAseq, qRT-PCR, H&E, and flow cytometry was used for analysis of colonic tissue/cell derived from B6ACTA2CreMyD88fl/fl, B6Col1a2CreMyD88fl/fl and B6RagCol1a2CreMyD88fl/fl MF-specific conditional KO mice. Results MF-specific deletion of MyD88 in vivo resulted in the development of microcolitis and was associated with dysbiotic decrease in Firmicutes to Bacteroidetes bacterial ratio, a hallmark of intestinal inflammation. This process was associated with the increase in colonic expression of genes linked to the inflammatory activity of myeloid cells and accumulation of immature monocyte-derived macrophages. In vitro deletion of MyD88 from MFs (isolated from MF-specific MyD88 conditional KO mice) resulted in changes of basal and TLR4 mediated pathways involved in myeloid cell migration/maturation and inflammation. Using MF-macrophage co-cultures we demonstrated the MF-restricted MyD88 signaling suppresses myeloid cell inflammatory activity. Conclusion Intrinsic MyD88 signaling within colonic mesenchymal cells is critical to control myeloid cell differentiation and inflammatory activity under mucosal tolerance in the gut. Supported by CTSA/TL1 5TL1TR001440-04 UTMB Institute for Translational Science and R01 DK103150 National Institute of Diabetes and Digestive and Kidney Diseases.

Inhibition of distinct glycolytic enzymes produces differential effects on CD4 T cell function

Abstract Background When T cells are activated, they upregulate glycolysis and take on a requisite Warburg phenotype. In this study, we evaluated the effect of inhibiting two distinct glycolytic enzymes, GAPDH and PGAM, on CD4 T cell differentiation and effector activities in order to gain a mechanistic understanding of the relationship between glycolysis and T cell function. Methods To evaluate the effect of GAPDH inhibition on T cell function, we cultured naïve murine CD4+ T cells under Th1 or Treg polarizing conditions and treated with either the GAPDH inhibitor heptelidic acid or the PGAM inhibitor EGCG. We then assessed either IFNγ production or identified CD25+Foxp3+ cells in response to either Th1 or Treg polarization, respectively. To evaluate the impact of GAPDH inhibition in vivo, we used the MOG35–55 experimental autoimmune encephalomyelitis (EAE) mouse model of multiple sclerosis (MS) and compared neurological function in mice treated with vehicle or heptelidic acid. Results We found that GAPDH inhibition had a potent anti-inflammatory effect on T cells in vitro, drastically reducing IFNγ production at both the transcriptional and protein levels. Interestingly, PGAM inhibition had the opposite effect, potently blocking Treg differentiation. In the EAE mouse model of MS, heptelidic acid attenuated the disease course, indicating that GAPDH inhibition is anti-inflammatory in vivo. Discussion GAPDH inhibition was anti-inflammatory in vitro and in vivo, while PGAM inhibition had the opposite effect. These findings suggest that the functional consequences of glycolysis inhibition depend on the specific enzymes targeted, which has both mechanistic and therapeutic implications for the role of glycolysis in immune cell function.

Genome-wide screening for genes involved in the epigenetic basis of fragile X syndrome.

SummaryFragile X syndrome (FXS), the most prevalent heritable form of intellectual disability, is caused by the transcriptional silencing of the FMR1 gene. The epigenetic factors responsible for FMR1 inactivation are largely unknown. Here, we initially demonstrated the feasibility of FMR1 reactivation by targeting a single epigenetic factor, DNMT1. Next, we established a model system for FMR1 silencing using a construct containing the FXS-related mutation upstream to a reporter gene. This construct was methylated in vitro and introduced into a genome-wide loss-of-function (LOF) library established in haploid human pluripotent stem cells (PSCs), allowing the identification of genes whose functional loss reversed the methylation-induced silencing of the FMR1 reporter. Selected candidate genes were further analyzed in haploid- and FXS-patient-derived PSCs, highlighting the epigenetic and metabolic pathways involved in FMR1 regulation. Our work sheds light on the mechanisms responsible for CGG-expansion-mediated FMR1 inactivation and offers novel targets for therapeutic FMR1 reactivation.

A network pharmacology technique used to investigate the potential mechanism of Ligustilide's effect on atherosclerosis.

Ligustilide (LIG) is a major active ingredient in traditional Chinese medicines that is also found in plant rhizomes such as carrot, coriander, and others, and it has been demonstrated to have cardiovascular preventive benefits. However, the mechanisms through which LIG protects the cardiovascular and cerebrovascular systems in atherosclerosis (AS) remain unknown. This study was aimed to investigate the mechanisms of LIG in AS utilizing the network pharmacology and molecular docking, and then to validate the putative mechanism through experiments. The network pharmacological analysis indicated that a total of 55 were performed on LIG and AS intersection targets. The genes of LIG and AS intersection targets enriched in the regulation of receptor and enzyme activity, cytokines-related, and transcription factors, indicating that these targets were primarily involved in cell proliferation and migration, regulating cell differentiation and skeletal activities in the development of AS. Finally, molecular docking was used to validate the major targets of LIG and AS intersection targets. Further experiments revealed that LIG may inhibit cell migration induced by AngII by reducing calcium influx, and regulating phenotypic translation-related proteins SM-22α and OPN. The present study investigated the potential targets and signaling pathways of LIG, which provides new insight into its anti-atherosclerosis actions in terms of reducing inflammation, cell proliferation, and migration, and may constitute a novel target for the treatment of AS. PRACTICAL APPLICATIONS: LIG has been shown to have cardiovascular protective benefits, the mechanism by which it protects the cardiovascular and cerebrovascular systems in AS remains unknown. This study uses a holistic network pharmacology strategy to investigate putative treatment pathways and conducts exploratory experimentation. The findings demonstrate that LIG reduces VSMC migration in the treatment of AS, acts as an anti-inflammatory agent, and prevents excessive cell proliferation and migration. Finally, the goal of our research is to uncover the molecular mechanism of LIG's influence on AS. The findings will provide a new research avenue for LIG as well as suggestions for the study of other herbal treatments. These research results will provide a new research direction for LIG and provide guidance for the research of other herbal medicines. This work revealed the multi-component, multi-target, multi-pathway, and multi-disease mechanism of LIG.

Tumor-derived extracellular vesicles induce invalid cytokine release and exhaustion of CD19 CAR-T Cells

Although CD19 chimeric antigen receptor-T (CAR-T) cells therapy has achieved unparalleled success in B cell malignancies. The dysfunction of CAR-T cells due to exhaustion is considered as a key factor for treatment failure, and the mechanisms of exhaustion remain elusive. Extracellular vesicles (EVs), important media for communication between tumor and immune cells, may contribute to CAR-T cell exhaustion. Here, we demonstrated that CD19+ tumor cells derived EVs (NALM6-EVs) can carry CD19 antigen and activate CD19 CAR-T cells. The transient activation induced a supraphysiologic inflammatory state with increased release of multiple cytokines. Besides, the sustained activation led CD19 CAR-T cells to enter an exhausted state with upregulated inhibitory receptors, decreased expansion ability, exaggerated effector cell differentiation and impaired antitumor activity. Transcriptomic profiling validated these findings and identified dynamic changes in CD8+ effector T, CD8+ exhausted T, CD8+RRM2+ T and T helper cell subpopulations during activation to exhaustion, as well as changes in many cytokines, inflammatory and immune-related pathways. Our findings identify a credible mechanism of CAR-T cell exhaustion that driven by tumor-derived EVs and provide a novel possible trigger for early cytokine release syndrome.

HIPK2 directs cell type–specific regulation of STAT3 transcriptional activity in Th17 cell differentiation

SignificanceSTAT3 (signal transducer and activator of transcription 3) is a master transcription factor that organizes cellular responses to cytokines and growth factors and is implicated in inflammatory disorders. STAT3 is a well-recognized therapeutic target for human cancer and inflammatory disorders, but how its function is regulated in a cell type-specific manner has been a major outstanding question. We discovered that Stat3 imposes self-directed regulation through controlling transcription of its own regulator homeodomain-interacting protein kinase 2 (Hipk2) in a T helper 17 (Th17) cell-specific manner. Our validation of the functional importance of the Stat3-Hipk2 axis in Th17 cell development in the pathogenesis of T cell-induced colitis in mice suggests an approach to therapeutically treat inflammatory bowel diseases that currently lack a safe and effective therapy.

ISuc-ChiDT: a computational method for identifying succinylation sites using statistical difference table encoding and the chi-square decision table classifier

BackgroundLysine succinylation is a type of protein post-translational modification which is widely involved in cell differentiation, cell metabolism and other important physiological activities. To study the molecular mechanism of succinylation in depth, succinylation sites need to be accurately identified, and because experimental approaches are costly and time-consuming, there is a great demand for reliable computational methods. Feature extraction is a key step in building succinylation site prediction models, and the development of effective new features improves predictive accuracy. Because the number of false succinylation sites far exceeds that of true sites, traditional classifiers perform poorly, and designing a classifier to effectively handle highly imbalanced datasets has always been a challenge.ResultsA new computational method, iSuc-ChiDT, is proposed to identify succinylation sites in proteins. In iSuc-ChiDT, chi-square statistical difference table encoding is developed to extract positional features, and has a higher predictive accuracy and fewer features compared to common position-based encoding schemes such as binary encoding and physicochemical property encoding. Single amino acid and undirected pair-coupled amino acid composition features are supplemented to improve the fault tolerance for residue insertions and deletions. After feature selection by Chi-MIC-share algorithm, the chi-square decision table (ChiDT) classifier is constructed for imbalanced classification. With a training set of 4748:50,551(true: false sites), ChiDT clearly outperforms traditional classifiers in predictive accuracy, and runs fast. Using an independent testing set of experimentally identified succinylation sites, iSuc-ChiDT achieves a sensitivity of 70.47%, a specificity of 66.27%, a Matthews correlation coefficient of 0.205, and a global accuracy index Q9 of 0.683, showing a significant improvement in sensitivity and overall accuracy compared to PSuccE, Success, SuccinSite, and other existing succinylation site predictors.ConclusionsiSuc-ChiDT shows great promise in predicting succinylation sites and is expected to facilitate further experimental investigation of protein succinylation.

Interplay between Dysbiosis of Gut Microbiome, Lipid Metabolism, and Tumorigenesis: Can Gut Dysbiosis Stand as a Prognostic Marker in Cancer?

The gut bacterial community is involved in the metabolism of bile acids and short-chain fatty acids (SCFAs). Bile acids are involved in the absorption of fat and the regulation of lipid homeostasis through emulsification and are transformed into unconjugated bile acids by the gut microbiota. The gut microbiota is actively involved in the production of bile acid metabolites, such as deoxycholic acid, lithocholic acid, choline, and SCFAs such as acetate, butyrate, and propionate. Metabolites derived from the gut microbiota or modified gut microbiota metabolites contribute significantly to host pathophysiology. Gut bacterial metabolites, such as deoxycholic acid, contribute to the development of hepatocellular carcinoma and colon cancer by factors such as inflammation and oxidative DNA damage. Butyrate, which is derived from gut bacteria such as Megasphaera, Roseburia, Faecalibacterium, and Clostridium, is associated with the activation of Treg cell differentiation in the intestine through histone acetylation. Butyrate averts the action of class I histone deacetylases (HDAC), such as HDAC1 and HDAC3, which are responsible for the transcription of genes such as p21/Cip1, and cyclin D3 through hyperacetylation of histones, which orchestrates G1 cell cycle arrest. It is essential to identify the interaction between the gut microbiota and bile acid and SCFA metabolism to understand their role in gastrointestinal carcinogenesis including colon, gastric, and liver cancer. Metagenomic approaches with bioinformatic analyses are used to identify the bacterial species in the metabolism of bile acids and SCFAs. This review provides an overview of the current knowledge of gut microbiota-derived bile acid metabolism in tumor development and whether it can stand as a marker for carcinogenesis. Additionally, this review assesses the evidence of gut microbiota-derived short-chain fatty acids including butyric acid in antitumor activity. Future research is required to identify the beneficial commensal gut bacteria and their metabolites which will be considered to be therapeutic targets in inflammation-mediated gastrointestinal cancers.

Synthesis of C2-Alkoxy-Substituted 19-Nor Vitamin D3 Derivatives: Stereoselectivity and Biological Activity

The active form of vitamin D3 (D3), 1a,25-dihydroxyvitamn D3 (1,25D3), plays a central role in calcium and bone metabolism. Many structure–activity relationship (SAR) studies of D3 have been conducted, with the aim of separating the biological activities of 1,25D3 or reducing its side effects, such as hypercalcemia, and SAR studies have shown that the hypercalcemic activity of C2-substituted derivatives and 19-nor type derivatives is significantly suppressed. In the present paper, we describe the synthesis of 19-nor type 1,25D3 derivatives with alkoxy groups at C2, by means of the Julia–Kocienski type coupling reaction between a C2 symmetrical A ring ketone and a CD ring synthon. The effect of C2 substituents on the stereoselectivity of the coupling reaction was evaluated. The biological activities of the synthesized derivatives were evaluated in an HL-60 cell-based assay. The a-methoxy-substituted C2α-7a was found to show potent cell-differentiating activity, with an ED50 value of 0.38 nM, being 26-fold more potent than 1,25D3.

The role of alpha-fetoprotein in the regulation of differentiation and functional activity of myeloid-derived suppressor cells

Myeloid-derived suppressor cells (MDSC) are innate immune cells that suppress the immune response. MDSC levels increase in pathological conditions as well as during pregnancy. Alpha-fetoprotein (AFP) is synthesized by embryonic tissues and then enters the mother's bloodstream, where it performs both transport and immunomodulatory functions. However, its role in regulating MDSC development has not been studied. In this work, the effect of physiological concentrations of recombinant AFP (10, 50, 100 IU/mL) on the differentiation of human MDSC in the in vitro system was investigated. To work with MDSC, an experimental model was developed to obtain sufficient numbers of these cells by cytokine induction of myeloid cells (CD33+). It was found that AFP at concentrations of 50 and 100 IU/ml increased the total number of MDSC in culture. The detailed analysis subpopulations revealed that the increase was due to monocytic M-MDSC. Thus, for the first time, the direct effect of recombinant AFP on MDSC differentiation have been demonstrated for the first time, which makes it possible in the future to formulate a new concept for its action as a pharmacological drug.

Hub Genes of Recurrent Laryngeal Cancer Identified by Weighted Gene Co-Expression Network Analysis

Objective in order to discover correlated gene modules and hub genes for recurrent laryngeal squamous cell carcinoma through weighted gene co-expression network analysis method. The microarray dataset of recurrent laryngeal cancer, namely GSE27020, were obtained from the gene expression Omnibus database. Weighted gene co-expression network analysis was introduced to establish a gene co-expression network, mining key clinical trait correlated hub genes. Gene ontology enrichment analyses were performed for the genes in modules related to recurrent laryngeal squamous cell carcinoma. Then, we build up a proteinprotein interaction network with the genes in interest gene module and identified hub genes through analyze such protein-protein interaction network. The hub genes were mined using cytohubba plus-in. Finally, we analyzed these hub genes overall survival using gene expression profiling interactive analysis database. Forty four gene co-expression gene modules were achieved via weighted gene co-expression network analysis analysis. We found that the orangered4 module was the most correlated module with recurrence in laryngeal squamous cell carcinoma patients. Genes in the orangered4 module were related to organelle organization, response to chemical, regulation of catalytic activity and regulation of cell differentiation. Two genes were discovered as hub genes which were related to poor prognosis, namely annexin A2 and S100 calcium binding protein A10. Here, we found several hub genes that played important roles in recurrent laryngeal squamous cell carcinoma, which may improve our understanding of the mechanisms underlying recurrence laryngeal squamous cell carcinoma.

Preparation and Properties of Decellularized Sheep Kidney Derived Matrix Scaffolds

Scaffolds from tissues or organs have nanoscale microstructures. Derived matrix scaffolds prepared by decellularized method can provide more cell attachment sites, which is conducive to cell adhesion, proliferation, differentiation and other physiological activities on scaffolds. In this study, the sheep kidney decellularized matrix scaffold was prepared by the method of decellularization. Due to the poor mechanical properties of the decellularized matrix, the cross linking method was adopted to enhance its mechanical properties. The decellularization efficiency of sheep renal matrix scaffolds was observed by scanning electron microscopy and histological staining, and the biocompatibility of the scaffolds was investigated by inoculating adipose derived stem cells. It was found that the scaffold had good decellularization effect and good pore structure.

Potential Application of Low-Intensity Pulsed Ultrasound in Delaying Aging for Mice

Introduction: The low-intensity pulsed ultrasound (LIPUS) is one of the popular treatment modalities allowing to boost the proliferation, differentiation, and migratory activity of cells, which might be a powerful strategy for anti-aging. Seeking a novel setup for LIPUS would benefit the development of ultrasound therapeutics. Methods: Here, we proposed a novel underwater exposure setup of LIPUS. C57BL/6 mice were reared in the designated age-groups, which consisted of a middle-aged group (12–14 months) and an old-age group (20–23 months). The age-related changes of body composition, imbalance of energy supply and demand, imbalance of signal network maintaining internal stability, and representative phenotypes of neurodegeneration and neuroplasticity with the presence and absence of underwater LIPUS in middle-aged and aged groups were evaluated. Results: The results showed that there were obvious aging changes, imbalance of energy supply and demand, imbalance of signal network maintaining homeostasis, neurodegeneration, and damage of neural plasticity in the middle-aged and aged group with or without the LIPUS. Although middle-aged group and aged group responded differently to LIPUS, they mostly generated positive results in relieving bone loss, improving ovarian structure, regulated immune system, and enhanced endurance ability, which should have declined over age. Discussion: These findings indicate that underwater extracorporeal LIPUS exposure could be employed as single or combined anti-aging strategies that generated positive outcomes against the process of aging.