The focus of this review is to provide an overview of the current state of knowledge of molecular mechanisms/processes that control differentiation of vascular smooth muscle cells (SMC) during normal development and maturation of the vasculature, as well as how these mechanisms/processes are altered in vascular injury or disease. A major challenge in understanding differentiation of the vascular SMC is that this cell can exhibit a wide range of different phenotypes at different stages of development, and even in adult organisms the cell is not terminally differentiated. Indeed, the SMC is capable of major changes in its phenotype in response to changes in local environmental cues including growth factors/inhibitors, mechanical influences, cell-cell and cell-matrix interactions, and various inflammatory mediators. There has been much progress in recent years to identify mechanisms that control expression of the repertoire of genes that are specific or selective for the vascular SMC and required for its differentiated function. One of the most exciting recent discoveries was the identification of the serum response factor (SRF) coactivator gene myocardin that appears to be required for expression of many SMC differentiation marker genes, and for initial differentiation of SMC during development. However, it is critical to recognize that overall control of SMC differentiation/maturation, and regulation of its responses to changing environmental cues, is extremely complex and involves the cooperative interaction of many factors and signaling pathways that are just beginning to be understood. There is also relatively recent evidence that circulating stem cell populations can give rise to smooth muscle-like cells in association with vascular injury and atherosclerotic lesion development, although the exact role and properties of these cells remain to be clearly elucidated. The goal of this review is to summarize the current state of our knowledge in this area and to attempt to identify some of the key unresolved challenges and questions that require further study.

Molecular Regulation of Vascular Smooth Muscle Cell Differentiation in Development and Disease

Endothelial Cells in Physiology and in the Pathophysiology of Vascular Disorders

How does the coagulation protease thrombin regulate cellular behaviour? The protease-activated receptors (PARs) provide one answer. In concert with the coagulation cascade, these receptors provide an elegant mechanism linking mechanical information in the form of tissue injury or vascular leakage to cellular responses. Roles for PARs are beginning to emerge in haemostasis and thrombosis, inflammation, and perhaps even blood vessel development.

Thrombin signalling and protease-activated receptors

Adipose tissue, best known for its role in fat storage, can also suppress weight gain and metabolic disease through the action of specialized, heat-producing adipocytes. Brown adipocytes are located in dedicated depots and express constitutively high levels of thermogenic genes, whereas inducible 'brown-like' adipocytes, also known as beige cells, develop in white fat in response to various activators. The activities of brown and beige fat cells reduce metabolic disease, including obesity, in mice and correlate with leanness in humans. Many genes and pathways that regulate brown and beige adipocyte biology have now been identified, providing a variety of promising therapeutic targets for metabolic disease.

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Brown and beige fat: development, function and therapeutic potential

Compounds Currently in Phase II−III Clinical Trials of Major Pharmaceutical Companies: New Structural Trends and Therapeutic Areas Yu Zhou,† Jiang Wang,† Zhanni Gu,† Shuni Wang,† Wei Zhu,† Jose ́ Luis Aceña,*,‡,§ Vadim A. Soloshonok,*,‡,∥ Kunisuke Izawa,* and Hong Liu*,† †Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China ‡Department of Organic Chemistry I, Faculty of Chemistry, University of the Basque Country UPV/EHU, Paseo Manuel Lardizab́al 3, 20018 San Sebastiań, Spain Department of Organic Chemistry, Autońoma University of Madrid, Cantoblanco, 28049 Madrid, Spain IKERBASQUE, Basque Foundation for Science, María Díaz de Haro 3, 48013 Bilbao, Spain Hamari Chemicals Ltd., 1-4-29 Kunijima, Higashi-Yodogawa-ku, Osaka, Japan 533-0024

Next Generation of Fluorine-Containing Pharmaceuticals, Compounds Currently in Phase II–III Clinical Trials of Major Pharmaceutical Companies: New Structural Trends and Therapeutic Areas

Disconnected interacting 2 homolog A (DIP2A) gene is highly expressed in nervous system and respiratory system of developing embryos. However, genes regulated by Dip2a in developing brain and lung have not been systemically studied. Transcriptome of brain and lung in embryonic 19.5 day (E19.5) were compared between wild type and Dip2a-/- mice. Total RNAs were extracted from brain and lung of E19.5 embryos for RNA-Seq. Clean reads were mapped to mouse reference sequence (mm9) using Tophat and assembled into transcripts by Cufflinks. Edge R and DESeq were applied to identify differentially expressed genes (DEGs) and annotated under GO, COG, KEGG and TF. An average of 50 million reads per sample was mapped to the reference sequence. A total of 214 DEGs were detected in brain (82 up and 132 down) and 1900 DEGs in lung (1259 up and 641 down). GO enrichment analysis indicated that DEGs in both Brain and Lung were mainly enriched in biological processes ‘DNA-templated transcription and Transcription from RNA polymerase II promoter’, ‘multicellular organism development’, ‘cell differentiation’ and ‘apoptotic process’. In addition, COG classification showed that both were mostly involved in ‘Replication, Recombination and Repair’, ‘Signal transduction and mechanism’, ‘Translation, Ribosomal structure and Biogenesis’ and ‘Transcription’. KEGG enrichment analysis showed that brain was mainly enriched in ‘Thryoid cancer’ pathway whereas lung in ‘Complement and Coagulation Cascades’ pathway. Transcription factor (TF) annotation analysis identified Zinc finger domain containing (ZF) proteins were mostly regulated in lung and brain. Interestingly, study identified genes Skor2, Gpr3711, Runx1, Erbb3, Frmd7, Fut10, Sox11, Hapln1, Tfap2c and Plxnb3 from brain that play important roles in neuronal cell maturation, differentiation and survival; genes Hoxa5, Eya1, Ctsh, Erff1, Lama1, Lama2, Rspo2, Sox11, Spry4, Shh, Igf1 and Wnt7a from lung are important in lung development and morphogenesis. Expression levels of the candidate genes were validated by qRT-PCR. Genome wide transcriptional analysis using wild type and Dip2a knockout mice in brain and lung at embryonic day 19.5 (E19.5) provided a genetic basis of molecular function of these genes.

https://www.biorxiv.org/content/biorxiv/early/2019/03/01/563916.full.pdf

Transcriptome Profiling of Mouse Brain and Lung under Dip2A Regulation Using RNA-Sequencing

The invention provides an application of an EGR-1 (early growth response-1) gene in preparation of anti-cancer medicine, and particularly provides an application of the EGR-1 gene in preparation of medicine for resisting the bladder cancer. An over-expression vector of the EGR-1 gene is built to transfect tumor cells, and a result shows that mRNA (messenger ribonucleic acid) of the EGR-1 gene in each transfected tumor cell is remarkably up-regulated, the tumor cells become round, wall attachment performance becomes poor, the cell viability is decreased by 66.2%, and the cells are dead finally. According to the invention, one gene capable of inhibiting the tumor cells efficiently is obtained through screening, and basic data are provided for novel medicine target discovery and antitumor medicine design.

Application of EGR-1 (early growth response-1) gene in preparation of medicine for resisting bladder cancer

[This corrects the article DOI: 10.1371/journal.pone.0213702.].

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Correction: Transcriptome profiling of mouse brain and lung under Dip2a regulation using RNA-sequencing.

The invention provides a use of a c-Fos gene in preparation of anti-cancer drugs and especially provides a use of the c-Fos gene in preparation of drugs for resisting bladder cancer An over-expression vector of the c-Fos gene is constructed and is transferred into cancer cells A result shows that mRNA and protein levels of the c-Fos gene in the cancer cells are obviously improved; roundness of the cancer cells is improved and adherence of the cancer cells is reduced; and cancer cell activity is reduced by 662% and the cancer cells die finally The c-Fos gene has high cancer cell inhibition efficiency and provides base data for novel drug target discovery and anti-cancer drug design

Use of c-Fos gene in preparation of anti-cancer drugs

Most human genetic diseases arise from point mutations with G:C>A:T or T:A>C:G base changes and they represent nearly half of the pathogenic single-nucleotide polymorphisms (SNPs). Animal models for human genetic diseases are important in dissecting pathogenic mechanism, drug screening, and drug efficacy testing. Mouse models are mostly generated by traditional gene knockout that is costly and time-consuming. CRISPR/Cas9 is a recently developed system that is efficient and cost-effective in generating genetic deletion, insertion and point mutation. It has been widely used to generate mouse models with deletions and point mutations. But, size and location of deletions by CRISPR/Cas9 is unpredictable. Mouse models of point mutation are still the best human disease models which can precisely mimic human pathology. Cytidine base editor BE4 is a newly developed version of cytidine base editing system. It has a cytidine deaminase and two uracil glycosylase inhibitors fused to C terminus of Cas9n, A cas9 mutant with D10A amino acid change. BE4 enables direct conversion of cytidine (C) to uridine (U) in targeted bases of DNA sequence. But this system has never been tested in vivo. In this study, we have confirmed that BE4 system can introduce site-specific and single-base substitution with high precision and efficiency in mouse. The designed nonsense mutation has a high efficiency up to 56.25%. Results confirm BE4 system has great potentials in modeling human genetic diseases and pharmaceutical screenings.

Yaowu Zheng

Papers

Transcriptome Profiling of Mouse Brain and Lung under Dip2A Regulation Using RNA-Sequencing

Application of EGR-1 (early growth response-1) gene in preparation of medicine for resisting bladder cancer

Correction: Transcriptome profiling of mouse brain and lung under Dip2a regulation using RNA-sequencing.

Use of c-Fos gene in preparation of anti-cancer drugs

Efficient Single Base Editing in Mouse Using Cytosine Base Editor 4