The Great Lakes Exposition

Held in the summers of 1936 and 1937, the Great Lakes Exposition was sprawled over 135 acres of land near Cleveland\u27s lakefront from W. 3rd St. to E. 20th St. Organized to celebrate the centennial year of the corporation of the city of Cleveland, the Great Lakes Exposition sought to highlight “the material, social and cultural progress which has been achieved in the Great Lakes Region in the past 100 year” and to “indicate the paths of progress for the future.”1 The Exposition featured hundreds of attractions, including rides, sideshows, botanical gardens, cafes, and more. In 1937, the Expo added an aquacade with water ballet shows featuring celebrities Johnny Weissmuller and Eleanor Holm. Some of the Cleveland/Northeast Ohio industries represented at the Expo included the White Motor Company, the Standard Oil Company, Firestone Tire & Rubber Company, Sherwin Williams, not to mention appearances by the Goodyear Tire & Rubber Co.\u27s ever-popular blimp. The Special Collections Department of the Michael Schwartz Library at Cleveland State University makes available a variety of resources that document the history and events of the Exposition. Many of the materials, including 150+ photographs and color postcards have been digitized and are available for viewing online. So take a step back in time and enjoy the sights of what was, without question, one of the most remarkable and beloved events in Cleveland history… line. 1Great Lakes Exposition Official Souvenir Guide, 1936.https://engagedscholarship.csuohio.edu/cmpex/1006/thumbnail.jp

The Great Lakes Exposition

Held in the summers of 1936 and 1937, the Great Lakes Exposition was sprawled over 135 acres of land near Cleveland\u27s lakefront from W. 3rd St. to E. 20th St. Organized to celebrate the centennial year of the corporation of the city of Cleveland, the Great Lakes Exposition sought to highlight “the material, social and cultural progress which has been achieved in the Great Lakes Region in the past 100 year” and to “indicate the paths of progress for the future.”1 The Exposition featured hundreds of attractions, including rides, sideshows, botanical gardens, cafes, and more. In 1937, the Expo added an aquacade with water ballet shows featuring celebrities Johnny Weissmuller and Eleanor Holm. Some of the Cleveland/Northeast Ohio industries represented at the Expo included the White Motor Company, the Standard Oil Company, Firestone Tire & Rubber Company, Sherwin Williams, not to mention appearances by the Goodyear Tire & Rubber Co.\u27s ever-popular blimp. The Special Collections Department of the Michael Schwartz Library at Cleveland State University makes available a variety of resources that document the history and events of the Exposition. Many of the materials, including 150+ photographs and color postcards have been digitized and are available for viewing online. So take a step back in time and enjoy the sights of what was, without question, one of the most remarkable and beloved events in Cleveland history… line. 1Great Lakes Exposition Official Souvenir Guide, 1936.https://engagedscholarship.csuohio.edu/cmpex/1006/thumbnail.jp

Using omics approaches to understand pulmonary diseases

Abstract Omics approaches are high-throughput unbiased technologies that provide snapshots of various aspects of biological systems and include: 1) genomics, the measure of DNA variation; 2) transcriptomics, the measure of RNA expression; 3) epigenomics, the measure of DNA alterations not involving sequence variation that influence RNA expression; 4) proteomics, the measure of protein expression or its chemical modifications; and 5) metabolomics, the measure of metabolite levels. Our understanding of pulmonary diseases has increased as a result of applying these omics approaches to characterize patients, uncover mechanisms underlying drug responsiveness, and identify effects of environmental exposures and interventions. As more tissue- and cell-specific omics data is analyzed and integrated for diverse patients under various conditions, there will be increased identification of key mechanisms that underlie pulmonary biological processes, disease endotypes, and novel therapeutics that are efficacious in select individuals. We provide a synopsis of how omics approaches have advanced our understanding of asthma, chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), idiopathic pulmonary fibrosis (IPF), and pulmonary arterial hypertension (PAH), and we highlight ongoing work that will facilitate pulmonary disease precision medicine

PDE8 Is Expressed in Human Airway Smooth Muscle and Selectively Regulates cAMP Signaling by β2-Adrenergic Receptors and Adenylyl Cyclase 6

Two cAMP signaling compartments centered on adenylyl cyclase (AC) exist in human airway smooth muscle (HASM) cells, one containing β2-adrenergic receptor AC6 and another containing E prostanoid receptor AC2. We hypothesized that different PDE isozymes selectively regulate cAMP signaling in each compartment. According to RNA-sequencing data, 18 of 24 PDE genes were expressed in primary HASM cells derived from age- and sex-matched donors with and without asthma. PDE8A was the third most abundant of the cAMP-degrading PDE genes, after PDE4A and PDE1A. Knockdown of PDE8A using shRNA evoked twofold greater cAMP responses to 1 μM forskolin in the presence of 3-isobutyl-1-methylxanthine. Overexpression of AC2 did not alter this response, but overexpression of AC6 increased cAMP responses an additional 80%. We examined cAMP dynamics in live HASM cells using a fluorescence sensor. PF-04957325, a PDE8-selective inhibitor, increased basal cAMP concentrations by itself, indicating a significant basal level of cAMP synthesis. In the presence of an AC inhibitor to reduce basal signaling, PF-04957325 accelerated cAMP production and increased the inhibition of cell proliferation induced by isoproterenol, but it had no effect on cAMP concentrations or cell proliferation regulated by prostaglandin E2. Lipid raft fractionation of HASM cells revealed PDE8A immunoreactivity in buoyant fractions containing caveolin-1 and AC5/6 immunoreactivity. Thus, PDE8 is expressed in lipid rafts of HASM cells, where it specifically regulates β2-adrenergic receptor AC6 signaling without effects on signaling by the E prostanoid receptors 2/4-AC2 complex. In airway diseases such as asthma and chronic obstructive pulmonary disease, PDE8 may represent a novel therapeutic target to modulate HASM responsiveness and airway remodeling

Rapamycin-independent <i>IGF2</i> expression in <i>Tsc2</i>-null mouse embryo fibroblasts and human lymphangioleiomyomatosis cells

<div><p>Lymphangioleiomyomatosis (LAM) is a rare, almost exclusively female lung disease linked to inactivating mutations in <i>tuberous sclerosis complex 2</i> (<i>TSC2)</i>, a tumor suppressor gene that controls cell metabolic state and growth via regulation of the mechanistic target of rapamycin (mTORC1) signaling. mTORC1 is frequently activated in human cancers and, although the mTORC1 inhibitor rapamycin has a cytostatic effect, it is, in general, unable to elicit a robust curative effect or tumor regression. Using RNA-Seq, we identified (1) <i>Insulin-like Growth Factor</i> (<i>IGF2</i>) as one of the genes with the highest fold-change difference between human <i>TSC</i>2-null and <i>TSC</i>2-expressing angiomyolipoma cells from a patient with LAM, and (2) the mouse <i>IGF2</i> homolog <i>Igf2</i>, as a top-ranking gene according to fold change between <i>Tsc</i>2^-/- and <i>Tsc</i>2^+/+ mouse embryo fibroblasts (MEFs). We extended transcript-level findings to protein level, observing increased Igf2 protein expression and Igf2 secretion by <i>Tsc</i>2^-/- MEFs. Increased Igf2 expression was not due to epigenetic imprinting, but was partially mediated through the Stat3 pathway and was completely insensitive to rapamycin treatment. An siRNA-mediated decrease of Igf2 resulted in decreased Stat3 phosphorylation, suggesting presence of an autocrine Igf2/Stat3 amplification cycle in <i>Tsc2</i>^<i>-/-</i> MEFs. In human pulmonary LAM lesions and metastatic cell clusters, high levels of IGF2 were associated with mTORC1 activation. In addition, treatment of three primary IGF2-expressing LAM lung cell lines with rapamycin did not result in IGF2 level changes. Thus, targeting of IGF2 signaling may be of therapeutic value to LAM patients, particularly those who are unresponsive to rapamycin.</p></div

Increased expression of <i>IGF2</i> transcripts in TSC2— human LAM cells and <i>Tsc2</i>^<i>-/-</i> MEFs.

<p>(A) TSC2 levels in human TSC2-null LAM 621–102 cells (TSC2—) cells and TSC2 re-expressing 621–103 LAM (TSC2++) cells. (B) RNA-Seq results show increased <i>IGF2</i> transcripts per kilobase million (TPM) in TSC2— cells. (C) Corresponding plot of mapped reads along the hg38 reference genome corresponding to <i>IGF2</i>. (D) Verification that <i>Tsc2</i> is not expressed in <i>Tsc2</i>^<i>-/-</i> MEFs. (E) RNA-Seq results show increased <i>Igf2</i> TPMs in <i>Tsc2</i>^<i>-/-</i> vs. <i>Tsc2</i>^<i>+/+</i> MEFs. (F) Corresponding plot of mapped reads along the mm10 reference genome corresponding to <i>Igf2</i>.</p

STAT3-dependent upregulation of IGF2 in TSC2-null cells.

<p>(A) Re-expression of TSC2 (TSC2++) in TSC2-null LAM 102 (TSC2—) cells decreased STAT3 expression and activation. (B) siRNA-induced knockdown of STAT3 decreased STAT3 levels in TSC2— cells. (C) RNA-Seq results show upregulated <i>IGF2</i> transcripts per kilobase million (TPM) in TSC2— cells transfected with either NT siRNA (Control) or STAT3 siRNA (siSTAT3). (D) STAT3 binding sites in human IGF2 and mouse Igf2 promoter regions. STAT3 enrichment in specific promoter regions of (E) human <i>IGF2</i> and (F) mouse <i>Igf2</i> genes was detected by ChIP-qPCR. Treatment of <i>Tsc2</i>^<i>-/-</i> MEFs with Stat3 inhibitor S3I-201 (100 nM for 18 hr) decreased Igf2 protein (G) expression as measured via Western blot and (H) secretion as measured via ELISA. (I) siRNA-mediated Stat3 knockdown also decreased Igf2 protein expression in <i>Tsc2</i>^<i>-/-</i> MEFs. (J) siRNA-mediated Igf2 knockdown decreased Stat3 phosphorylation but not total Stat3.</p

IGF2 expression in LAM lungs and <i>Tsc2</i>^<i>-/-</i> MEFs.

<p>Representative images of IHC analysis show IGF2 expression in (A) LAM lesion and (B) LAM cluster detected with specific antibodies. Non-immune IgG was used as a control (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0197105#pone.0197105.s001" target="_blank">S1 Fig</a>). Igf2 expression in <i>Tsc2</i>^<i>-/-</i> MEFs was detected by (C) qPCR (D) Western blot and (E) ELISA. (F) <i>Tsc2</i>^<i>-/-</i> MEFs were transfected with 50nM <i>Igf2</i> siRNA (siIGF2) or NT siRNA (siNT) for 48 hrs. Decreased levels of Igf2 protein expression were confirmed via western blot with β-actin as an internal loading control. (G) Decreased Igf2 protein secretion was confirmed via ELISA. Igf2 knockdown resulted in (H) increased cleaved caspase-3 levels as measured via immunocytostaining and flow for Alexa Fluor® 488 -Cleaved Caspase 3 where the population of positively stained MEFs was normalized to the control population, and (J) decreased cell viability as assessed by 0.4% Trypan Blue staining normalized to the control cell viability. Student's t-tests were used to determine the statistical significance of the differences, and <i>p</i>-values reflect a sample size of 3 replicates.</p