DOI: 10.5061/dryad.n02v6wx7f
title: Endothelial progenitor cell gene expression fold-changes in filtered air versus CAP-exposed animals

description: While some prior studies have identified an association between exposure
 to fine air-borne particulate matter (PM2.5) and indices of aging, the
 extent of these associations and their underlying mechanisms are
 uncertain. In this study, we exposed male C57BL/6J mice to filtered air
 and concentrated ambient PM2.5 (CAP) and assessed two common hallmarks of
 aging, telomere length and senescence. Of the cell types examined,
 peripheral blood mononuclear cells (PBMNCs), endothelial progenitor cells
 (EPCs), and bone marrow-derived c-kit+ cells all demonstrated shortened
 telomeres when isolated from CAP-exposed as compared with cells from
 filtered air controls. Telomere attrition in PBMNCs and EPCs resulted from
 the attenuated catalytic activity of telomerase reverse transcriptase
 (Tert). We found that telomere attrition in these cell types was mitigated
 in those CAP-exposed mice receiving water supplemented with the
 antioxidant, carnosine. However, telomere attrition was reversible in
 PBMNCs, but not EPCs, when CAP-exposed mice were allowed to recover in
 normal air conditions. PBMNCs and EPCs obtained from CAP-exposed mice****
 also displayed increased β-galactosidase activity and expression of genes
 characteristic of the senescence-activated secretory phenotype. PBMNC
 senescence was greatest in CD8+ T-cells. Our results suggest that the
 pro-aging effects of PM2.5 impact multiple cell types, including bone
 marrow stem cells, and that telomere attrition resulted from attenuated
 Tert activity. The aging and senescence of multiple cell types, including
 bone marrow stem cells, may underlie the diverse pathological outcomes of
 PM2.5 exposure.
description: We used RT-PCR to analyze expression levels of telomerase
 reserves transcriptase (<em>Tert</em>), genes encoding
 telomerase-associated proteins, proteins of the shelterin complex, and
 genes indicative of senescence. RNA was isolated from frozen cell pellets
 using the miRNeasy isolation kit (Qiagen, #217004) and concentrations
 determined on a Nanodrop One spectrophotometer. 100ng of RNA was then
 reverse transcribed with random primers using a High-Capacity cDNA Reverse
 Transcription Kit (Thermo Fisher #4368814). Finally, we used specific
 TaqMan primers (Thermo Fisher) and the TaqMan 2x Universal PCR Master Mix
 (Thermo Fisher, #4324018) to amplify sequences encoding
 <em>Tert</em> (Mm00436931_m1), <em>Dkc1</em>
 (Mm01222888_m1), <em>Nop10</em> (Mm00777618_g1),
 <em>Nhp2</em> (Mm00835429_g1), <em>Gar1</em>
 (Mm00452760_g1), <em>Terf1</em> (Mm­00436928_m1),
 <em>Terf2</em> (mm01253555_m1), <em>Terf2ip</em>
 (Mm01243676_m1), <em>Pinx1</em> (Mm00499603_m1), 
 <em>Tin2</em> (Mm00461166_g1), <em>Tpp1</em>
 (Mm03989838_g1), <em>p16<sup>INK4A</sup></em>
 (Mm00494449_m1), <em>Ccl5</em> (Mm01302427_m1),
 <em>Tnfa</em> (Mm00443258_m1) and <em>IL-1b</em>
 (Mm00434228_m1). <em>Hprt</em> primers (Mm03024075_m1) were
 used to amplify an internal control. Calculations to determine expression
 levels were done using the DDCt method (Livak and Schmittgen
 2001)
description: # Endothelial progenitor cell gene expression fold-changes in filtered air
 versus CAP-exposed animals
 [https://doi.org/10.5061/dryad.n02v6wx7f](https://doi.org/10.5061/dryad.n02v6wx7f) ## Description of the data and file structure One goal of the study was to determine if murine exposure to concentrated ambient air pollution particles (CAP) altered the expression of telomere-associated proteins in endothelial progenitor cells (EPCs) isolated and cultured from these mice. Mice exposed to filtered air were used as controls. **Gene expression levels were determined by RT-PCR, and results were expressed as a fold change**. #### Description of the data and file structure Data is listed as a fold-change in CAP vs. filtered air. The columns (gene names) of the tel_rtPCR_data_for_Dryad_(1).csv are as follows: * Tert: telomerase reverse transcriptase - Dkc1: Dyskerin * Gar1: Nucleolar Protein Family A, Member 1 - Nhp2: Nucleolar Protein Family A, Member 2 * Nop10: Nucleolar Protein Family A, Member 3 - Terf1: Telomeric Repeat Binding Factor 1 * Terf2: Telomeric Repeat Binding Factor 2 - Terf2ip: Telomeric Repeat-Binding Factor 2-Interacting Protein 1 * Pinx1: PIN2/TERF1-Interacting Telomerase Inhibitor 1 - Tpp1: ACD Shelterin Complex Subunit And Telomerase Recruitment Factor * Tin2: TERF1 Interacting Nuclear Factor