Identification of the early and late responder genes during the generation of induced pluripotent stem cells from mouse fibroblasts

111Ysciescopu

Long-term efficacy, safety and immunogenicity in patients with rheumatoid arthritis continuing on an etanercept biosimilar (LBEC0101) or switching from reference etanercept to LBEC0101: an open-label extension of a phase III multicentre, randomised, double-blind, parallel-group study

Background To evaluate the long-term efficacy, safety and immunogenicity of continuing LBEC0101; the etanercept (ETN) biosimilar; or switching from the ETN reference product (RP) to LBEC0101 in patients with rheumatoid arthritis (RA). Methods This multicentre, single-arm, open-label extension study enrolled patients who had completed a 52-week randomised, double-blind, parallel phase III trial of LBEC0101 vs ETN-RP. Patients treated with ETN-RP during the randomised controlled trial switched to LBEC0101; those treated with LBEC0101 continued to receive LBEC0101 in this study. LBEC0101 (50 mg) was administered subcutaneously once per week for 48 weeks with a stable dose of methotrexate. Efficacy, safety and immunogenicity of LBEC0101 were assessed up to week 100. Results A total of 148 patients entered this extension study (70 in the maintenance group and 78 in the switch group). The 28-joint disease activity scores (DAS28)-erythrocyte sedimentation rate (ESR) were maintained in both groups from week 52 to week 100 (from 3.068 to 3.103 in the maintenance group vs. from 3.161 to 3.079 in the switch group). ACR response rates at week 100 for the maintenance vs. switch groups were 79.7% vs. 83.3% for ACR20, 65.2% vs. 66.7% for ACR50 and 44.9% vs. 42.3% for ACR70. The incidence of adverse events and the proportion of patients with newly developed antidrug antibodies were similar in the maintenance and switch groups (70.0% and 70.5%, 1.4% and 1.3%, respectively). Conclusions Administration of LBEC0101 showed sustained efficacy and acceptable safety in patients with RA after continued therapy or after switching from ETN-RP to LBEC0101. Trial registration ClinicalTrials.gov, NCT02715908. Registered 22 March 2016.This extension study was funded by LG Chem, Ltd. (formerly, LG Life Sciences, Ltd), Mochida Pharmaceutical Co., Ltd. and Korea Health Industry Development Institute

Post-stroke dementia - a comprehensive review

Post-stroke dementia (PSD) or post-stroke cognitive impairment (PSCI) may affect up to one third of stroke survivors. Various definitions of PSCI and PSD have been described. We propose PSD as a label for any dementia following stroke in temporal relation. Various tools are available to screen and assess cognition, with few PSD-specific instruments. Choice will depend on purpose of assessment, with differing instruments needed for brief screening (e.g., Montreal Cognitive Assessment) or diagnostic formulation (e.g., NINDS VCI battery). A comprehensive evaluation should include assessment of pre-stroke cognition (e.g., using Informant Questionnaire for Cognitive Decline in the Elderly), mood (e.g., using Hospital Anxiety and Depression Scale), and functional consequences of cognitive impairments (e.g., using modified Rankin Scale). A large number of biomarkers for PSD, including indicators for genetic polymorphisms, biomarkers in the cerebrospinal fluid and in the serum, inflammatory mediators, and peripheral microRNA profiles have been proposed. Currently, no specific biomarkers have been proven to robustly discriminate vulnerable patients (‘at risk brains’) from those with better prognosis or to discriminate Alzheimer’s disease dementia from PSD. Further, neuroimaging is an important diagnostic tool in PSD. The role of computerized tomography is limited to demonstrating type and location of the underlying primary lesion and indicating atrophy and severe white matter changes. Magnetic resonance imaging is the key neuroimaging modality and has high sensitivity and specificity for detecting pathological changes, including small vessel disease. Advanced multi-modal imaging includes diffusion tensor imaging for fiber tracking, by which changes in networks can be detected. Quantitative imaging of cerebral blood flow and metabolism by positron emission tomography can differentiate between vascular dementia and degenerative dementia and show the interaction between vascular and metabolic changes. Additionally, inflammatory changes after ischemia in the brain can be detected, which may play a role together with amyloid deposition in the development of PSD. Prevention of PSD can be achieved by prevention of stroke. As treatment strategies to inhibit the development and mitigate the course of PSD, lowering of blood pressure, statins, neuroprotective drugs, and anti-inflammatory agents have all been studied without convincing evidence of efficacy. Lifestyle interventions, physical activity, and cognitive training have been recently tested, but large controlled trials are still missing

Factors Associated with Long-Term Dietary Supplement Use among Korean Breast Cancer Survivors: A Cross-Sectional Study

Purpose: The factors associated with the dietary supplement (DS) use of Asian breast cancer survivors in consideration of the duration of use and types of DS have not been well established. Methods: We recruited 693 Korean female breast cancer survivors at two university-affiliated hospitals and collected study data through a self-administered questionnaire and a review of medical records. A multiple logistic regression analysis was conducted to evaluate the multivariable-adjusted association between DS use and study variables. Results: The prevalence of any (≥2 weeks) and long-term (≥6 months) DS use among study participants was 48.2% and 12.0%, respectively. Education level, alcohol use, adequate physical activity (≥150 min/week), and time lapse after cancer diagnosis were positively associated with any DS use. Among DS users, as compared with short-term (≥2 weeks and <6 months) users, long-term users were more likely to have a higher cancer stage, more diverse cancer treatment modalities, a shorter time since cancer diagnosis, and lower fear of cancer recurrence. When we repeated the analysis for each DS type, time lapse after cancer diagnosis showed a consistently inverse association with long-term use of the most frequently consumed DS (multivitamins, followed by vitamin D/calcium, vitamin C, and omega-3). The number of cancer treatment modalities was positively associated with the long-term use of multivitamins and vitamin D/calcium. Alcohol consumption and low bone mineral density were positively associated with long-term vitamin D/calcium use. Conclusions: The factors associated with DS use differed by the duration of DS use and specific DS type. Long-term DS use was more frequently associated with cancer-related factors

Identification of the early and late responder genes during the generation of induced pluripotent stem cells from mouse fibroblasts

<div><p>Background</p><p>The generation of induced pluripotent stem cell (iPSC), a substitute for embryonic stem cell (ESC), requires the proper orchestration of a transcription program at the chromatin level. Our recent approach for the induction of pluripotent stem cells from fibroblasts using protein extracts from mouse ESCs could overcome the potential tumorigenicity risks associated with random retroviral integration. Here, we examine the epigenetic modifications and the transcriptome of two types of iPSC and of partially reprogrammed iPSCs (iPSCp) generated independently from adult cardiac and skin fibroblasts to assess any perturbations of the transcription program during reprogramming.</p><p>Results</p><p>The comparative dissection of the transcription profiles and histone modification patterns at lysines 4 and 27 of histone H3 of the iPSC, iPSCp, ESC, and somatic cells revealed that the iPSC was almost completely comparable to the ESC, regardless of their origins, whereas the genes of the iPSCp were dysregulated to a larger extent. Regardless of the origins of the somatic cells, the fibroblasts induced using the ESC protein extracts appear to be completely reprogrammed into pluripotent cells, although they show unshared marginal differences in their gene expression programs, which may not affect the maintenance of stemness. A comparative investigation of the iPSCp generated by unwanted reprogramming showed that the two groups of genes on the pathway from somatic cells to iPSC might function as sequential reprogramming-competent early and late responders to the induction stimulus. Moreover, some of the divergent genes expressed only in the iPSCp were associated with many tumor-related pathways.</p><p>Conclusions</p><p>Faithful transcriptional reprogramming should follow epigenetic alterations to generate induced pluripotent stem cells from somatic cells. This genome-wide comparison enabled us to define the early and late responder genes during the cell reprogramming process to iPSC. Our results indicate that the cellular responsiveness to external stimuli should be pre-determined and sequentially orchestrated through the tight modulation of the chromatin environment during cell reprogramming to prevent unexpected reprogramming.</p></div

Characterization of epigenetic signatures related to gene expression in iPSCs.

<p>(A-F). The scatter plots show the fold changes in H3K4me3, H3K27me3, and gene expression during reprogramming. Each dot represents a gene with p-value less than 0.01. The X and Y axes indicate the fold changes in H3K4me3 and H3K27me3 (log2 scale), respectively. The color code shows the gene expression level from low to high (green-black-red). The percentage of differentially expressed genes is shown in the inset pie chart (ANOVA test p-value<0.01, fold change>2). (G) The chromatin states of the key transcription factor genes, such as the Oct4, Nanog, and Sox2, are shown on the UCSC genome browser.</p

Comparative analysis of the protein-based iPSCs and other pluripotent cells.

<p>(A) Hierarchical clustering of the gene expression microarray data for the mESCs (open circles), iPSCs (filled circles), and fibroblasts (filled squares) from different laboratories. The data sets of the mESCs and iPSCs obtained from the same laboratory are marked using grey boxes. The iPSC induction methods are labelled for each iPSC data set: ¹GSE13770, ²GSE24930, ³GSE17004, ⁴GSE27814, ⁵GSE22908, ⁶GSE24046, and ⁷GSE27087. (B-C) The Venn diagram shows the overlap of the differentially expressed genes between mESCs and iPSCs. For comparison, two iPSC* (GSE24046) and iPSC^§ (GSE27814) data sets are also incorporated. (D) The gene expression levels and histone modifications of the differentially expressed genes defined in (B-C) are shown and the Pearson correlation coefficients are presented on the right side of each heatmap.</p

Global histone modification signatures of pluripotent and somatic cells.

<p>(A) Experimental overview of the genome-wide analysis of the histone modifications of mESC, iPSCs and the original somatic cells using ChIP-Seq and their gene expression measured via microarray and RNA-seq. (B) The histone modification profiles of the Hox D cluster genes are shown using the UCSC genome browser. The genomic position of the region is indicated on top of the map. (C) Chromosome-wide H3K4me3 peak patterns are compared in different cells. The Hilbert plots of H3K4me3 enrichments at chromosome 1 are visualized using the HilberVis program. (D) The comparison of histone modification patterns between different mESCs and iPSCs. The heatmap represents the distributions of the histone modification near the TSS of 22,086 RefSeq genes. The rows in the all data sets are sorted using the tag density of the mESCs from the highest to the lowest. The position of the TSS and the direction of transcription are denoted using an arrow. The numbers inside the heatmap indicate the Pearson correlation coefficients compared with the mESCs.</p

Identification of early and late responder genes from the partial iPSC analysis.

<p>(A) Categorization of the differentially expressed genes among iPSCps, mESCs, and sFB-G into three groups: the divergent, the convergent, and the resistant genes. (B) The H3K4me3 and H3K27me3 levels were plotted, depending on the responsiveness of the genes to the induction of pluripotency (C) The UCSC genome browser shows the chromatin states of the divergent (Fgf5), convergent (Cdc20), and resistant (Col1a1) genes in all cell types. The positions and directions of transcription of the genes are indicated below the panel. (D) The three groups are listed according to their expression and histone modification fold change values. The gene ontology analysis shows highly significant biological functions associated with the differentially expressed genes. (E) A model for the step-wise acquisition of pluripotency. Sox2 was identified as an early responder gene.</p