ABCs of COVID-19 Coloring Page

To help give children a better understanding of the coronavirus that is causing the COVID-19 disease and the disruption in our daily lives, the RNA Therapeutics Institute (RTI) at UMass Medical created this coloring page just for kids. We hope parents and teachers can use it to help explain this complex situation, while also teaching the wonders of science.https://escholarship.umassmed.edu/rti_kids/1001/thumbnail.jp

Coronavirus Infographic

To help give children a better understanding of the coronavirus that is causing the COVID-19 disease and the disruption in our daily lives, the RNA Therapeutics Institute (RTI) at UMass Medical created this infographic just for kids. We hope parents and teachers can use it to help explain this complex situation, while also teaching the wonders of science.https://escholarship.umassmed.edu/rti_kids/1000/thumbnail.jp

Endothelial Differentiation Gene-1, a New Downstream Gene Is Involved in RTEF-1 Induced Angiogenesis in Endothelial Cells

Related Transcriptional Enhancer Factor-1 (RTEF-1) has been suggested to induce angiogenesis through regulating target genes. Whether RTEF-1 has a direct role in angiogenesis and what specific genes are involved in RTEF-1 driven angiogenisis have not been elucidated. We found that over-expressing RTEF-1 in Human dermal microvascular endothelial cells-1 (HMEC-1) significantly increased endothelial cell aggregation, growth and migration while the processes were inhibited by siRNA of RTEF-1. In addition, we observed that Endothelial differentiation gene-1 (Edg-1) expression was up-regulated by RTEF-1 at the transcriptional level. RTEF-1 could bind to Edg-1 promoter and subsequently induce its activity. Edg-1 siRNA significantly blocked RTEF-1-driven increases in endothelial cell aggregation in a Matrigel assay and retarded RTEF-1-induced endothelial cell growth and migration. Pertussis Toxin (PTX), a Gi/Go protein sensitive inhibitor, was found to inhibit RTEF-1 driven endothelial cell aggregation and migration. Our data demonstrates that Edg-1 is a potential target gene of RTEF-1 and is involved in RTEF-1-induced angiogenesis in endothelial cells. Gi/Go protein coupled receptor pathway plays a role in RTEF-1 driven angiogenesis in endothelial cells

Role of A20 in cIAP-2 Protection against Tumor Necrosis Factor α (TNF-α)-Mediated Apoptosis in Endothelial Cells

Tumor necrosis factor α (TNF-α) influences endothelial cell viability by altering the regulatory molecules involved in induction or suppression of apoptosis. However, the underlying mechanisms are still not completely understood. In this study, we demonstrated that A20 (also known as TNFAIP3, tumor necrosis factor α-induced protein 3, and an anti-apoptotic protein) regulates the inhibitor of apoptosis protein-2 (cIAP-2) expression upon TNF-α induction in endothelial cells. Inhibition of A20 expression by its siRNA resulted in attenuating expression of TNF-α-induced cIAP-2, yet not cIAP-1 or XIAP. A20-induced cIAP-2 expression can be blocked by the inhibition of phosphatidyl inositol-3 kinase (PI3-K), but not nuclear factor (NF)-κB, while concomitantly increasing the number of endothelial apoptotic cells and caspase 3 activation. Moreover, TNF-α-mediated induction of apoptosis was enhanced by A20 inhibition, which could be rescued by cIAP-2. Taken together, these results identify A20 as a cytoprotective factor involved in cIAP-2 inhibitory pathway of TNF-α-induced apoptosis. This is consistent with the idea that endothelial cell viability is dependent on interactions between inducers and suppressors of apoptosis, susceptible to modulation by TNF-α

IGF binding protein‐6 expression in vascular endothelial cells is induced by hypoxia and plays a negative role in tumor angiogenesis

Hypoxia stimulates tumor angiogenesis by inducing the expression of angiogenic molecules. The negative regulators of this process, however, are not well understood. Here, we report that hypoxia induced the expression of insulin‐like growth factor binding protein‐6 (IGFBP‐6), a tumor repressor, in human and rodent vascular endothelial cells (VECs) via a hypoxia‐inducible factor (HIF)‐mediated mechanism. Addition of human IGFBP‐6 to cultured human VECs inhibited angiogenesis in vitro . An IGFBP‐6 mutant with at least 10,000‐fold lower binding affinity for IGFs was an equally potent inhibitor of angiogenesis, suggesting that this action of IGFBP‐6 is IGF‐independent. The functional relationship between IGFBP‐6 and vascular endothelial growth factor (VEGF), a major hypoxia‐inducible angiogenic molecule, was examined. While VEGF alone increased angiogenesis in vitro , co‐incubation with IGFBP‐6 abolished VEGF‐stimulated angiogenesis. The in vivo role of IGFBP‐6 in angiogenesis was tested in flk1 :GFP zebrafish embryos, which exhibit green fluorescence protein in developing vascular endothelium, permitting visualization of developing blood vessels. Injection of human IGFBP‐6 mRNA reduced the number of embryonic inter‐segmental blood vessels by ∼40%. This anti‐angiogenic activity is conserved in zebrafish because expression of zebrafish IGFBP‐6b had similar effects. To determine the anti‐angiogenic effect of IGFBP‐6 in a tumor model, human Rh30 rhabdomyosarcoma cells stably transfected with IGFBP‐6 were inoculated into athymic BALB/c nude mice. Vessel density was 52% lower in IGFBP‐6‐transfected xenografts than in vector control xenografts. These results suggest that the expression of IGFBP‐6 in VECs is up‐regulated by hypoxia and IGFBP‐6 inhibits angiogenesis in vitro and in vivo .Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/90159/1/26201_ftp.pd

Determinants of GBP Recruitment to Toxoplasma gondii Vacuoles and the Parasitic Factors That Control It

IFN-γ is a major cytokine that mediates resistance against the intracellular parasite Toxoplasma gondii. The p65 guanylate-binding proteins (GBPs) are strongly induced by IFN-γ. We studied the behavior of murine GBP1 (mGBP1) upon infection with T. gondii in vitro and confirmed that IFN-γ-dependent re-localization of mGBP1 to the parasitophorous vacuole (PV) correlates with the virulence type of the parasite. We identified three parasitic factors, ROP16, ROP18, and GRA15 that determine strain-specific accumulation of mGBP1 on the PV. These highly polymorphic proteins are held responsible for a large part of the strain-specific differences in virulence. Therefore, our data suggest that virulence of T. gondii in animals may rely in part on recognition by GBPs. However, phagosomes or vacuoles containing Trypanosoma cruzi did not recruit mGBP1. Co-immunoprecipitation revealed mGBP2, mGBP4, and mGBP5 as binding partners of mGBP1. Indeed, mGBP2 and mGBP5 co-localize with mGBP1 in T. gondii-infected cells. T. gondii thus elicits a cell-autonomous immune response in mice with GBPs involved. Three parasitic virulence factors and unknown IFN-γ-dependent host factors regulate this complex process. Depending on the virulence of the strains involved, numerous GBPs are brought to the PV as part of a large, multimeric structure to combat T. gondii.National Institutes of Health (U.S.)Massachusetts Life Sciences Center (New Investigator Award)National Institute of General Medical Sciences (U.S.) (Pre-Doctoral Grant in the Biological Sciences (5-T32-GM007287-33))Studienstiftung des deutschen VolkesCancer Research Institute (New York, N.Y.)Cleo and Paul Schimmel FoundationBayer HealthcareHuman Frontier Science Program (Strasbourg, France

Intracellular Trafficking of Guanylate-Binding Proteins Is Regulated by Heterodimerization in a Hierarchical Manner

Guanylate-binding proteins (GBPs) belong to the dynamin family of large GTPases and represent the major IFN-γ-induced proteins. Here we systematically investigated the mechanisms regulating the subcellular localization of GBPs. Three GBPs (GBP-1, GBP-2 and GBP-5) carry a C-terminal CaaX-prenylation signal, which is typical for small GTPases of the Ras family, and increases the membrane affinity of proteins. In this study, we demonstrated that GBP-1, GBP-2 and GBP-5 are prenylated in vivo and that prenylation is required for the membrane association of GBP-1, GBP-2 and GBP-5. Using co-immunoprecipitation, yeast-two-hybrid analysis and fluorescence complementation assays, we showed for the first time that GBPs are able to homodimerize in vivo and that the membrane association of GBPs is regulated by dimerization similarly to dynamin. Interestingly, GBPs could also heterodimerize. This resulted in hierarchical positioning effects on the intracellular localization of the proteins. Specifically, GBP-1 recruited GBP-5 and GBP-2 into its own cellular compartment and GBP-5 repositioned GBP-2. In addition, GBP-1, GBP-2 and GBP-5 were able to redirect non-prenylated GBPs to their compartment in a prenylation-dependent manner. Overall, these findings prove in vivo the ability of GBPs to dimerize, indicate that heterodimerization regulates sub-cellular localization of GBPs and underscore putative membrane-associated functions of this family of proteins

COVID-19 mRNA vaccines: how they work and why they are safe

Get the facts of the COVID-19 mRNA vaccines. This infographic explains how the mRNA vaccines prevent severe disease from SARS-CoV-2, the virus causing COVID-19. These vaccines have been thoroughly tested and found to be safe and effective and continue to undergo continuous and intense safety monitoring. This infographic dispels some of the common myths associated with the vaccine.https://escholarship.umassmed.edu/rti_kids/1002/thumbnail.jp

Emma RNA Saves the Day

In 2019, a new and dangerous villain emerged in the world, Spike Man. Spike Man embodies the coronavirus, SARS-CoV-2, the virus that causes COVID-19. In ScienceLIVE’s first coloring comic book, you’ll learn about how Spike Man tries to hijack the body’s factories and how Emma RNA and her legion of molecular superheroes save the day! This coloring book is available in English, Spanish, French, Turkish, Chinese, Portuguese and German. Spanish description: En el 2019 un nuevo y peligroso villano apareció en el mundo, el Hombre Espiga. El Hombre Espiga representa el coronavirus, SARS-CoV-2, el cual causa el COVID-19. En el primer libro para colorear de ScienceLIVE, aprenderás como el Hombrer Epsiga trata de secuestra las fábricas del cuerpo, y como Emma ARN y su legion de superheroes moleculares salvan el día. French description: En 2019, un nouveau et très dangereux vilain a émergé sur la planète, l’homme spicule. L’homme spicule représente le coronavirus, SARS-CoV-2, le virus qui cause la COVID-19. Dans ce premier livre à colorier de ScienceLIVE, tu vas apprendre les tactiques utilisées par l’homme spicule pour détourner les usines du corps humain et comment Emma ARN et son armée de super-héros moléculaires sauvent la situation. Turkish description: 2019 yılında dünyada yeni ve tehlikeli bir kötü adam türedi, Sivri Adam. Sivri Adam, COVID-19 hastalığına neden olan SARS-CoV-2 adındaki koronavirüsü temsil ediyor. ScienceLIVE programının bu ilk boyama kitabında, Sivri Adam’ın nasıl vücudun fabrikalarını ele geçirmeye uğraştığını ve Melda RNA (mRNA aşısı) ve moleküler süperkahramanlar ekibinin vücudu korumak için iş başında olduğunu öğreneceksiniz.https://escholarship.umassmed.edu/rti_kids/1003/thumbnail.jp