Endothelial progenitor cell gene expression fold-changes in filtered air versus CAP-exposed animals
Data files
Nov 26, 2025 version files 2.29 KB
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README.md
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tel_rtPCR_data_for_Dryad_(1).csv
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Abstract
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.
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
We used RT-PCR to analyze expression levels of telomerase reserves transcriptase (Tert), 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 Tert (Mm00436931_m1), Dkc1 (Mm01222888_m1), Nop10 (Mm00777618_g1), Nhp2 (Mm00835429_g1), Gar1 (Mm00452760_g1), Terf1 (Mm00436928_m1), Terf2 (mm01253555_m1), Terf2ip (Mm01243676_m1), Pinx1 (Mm00499603_m1), Tin2 (Mm00461166_g1), Tpp1 (Mm03989838_g1), p16INK4A (Mm00494449_m1), Ccl5 (Mm01302427_m1), Tnfa (Mm00443258_m1) and IL-1b (Mm00434228_m1). Hprt 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)