biostudies-arrayexpressDavid JohnHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-17485Aging is a major risk factor for cardiovascular diseases, yet the contribution of the lymphatic vasculature to cardiac aging remains largely unexplored. Here, we show that aging reduces lymphatic vessel density in human and mice hearts and induces morphological changes, including the formation of zipper-like, tighter endothelial junctions. These alterations are accompanied by immune cell infiltration, fibrinogen and amyloid accumulation, and myocardial edema. Experimental reduction of cardiac lymphatics in young mice, achieved by Flt4 (VEGFR3) depletion or overexpression of soluble Flt4, reproduces some age-related cardiac phenotypes, such as inflammation and impaired lymphatic integrity. Mechanistically, we found that aging induces the selective up-regulation of nuclear interleukin 33 (IL33) in lymphatic endothelial cells. In contrast to the extracellular, cardioprotective form of IL33, nuclear IL33 promotes lymphatic cell death and junctional remodeling. A targeted screen of pro-lymphatic factors identified VEGFC as an age-sensitive regulator that both declines in the aging heart and suppresses IL33. Cardiac Vegfc overexpression or Il33 silencing restores lymphatic vessel density, reduces macrophage infiltration, and improves tissue homeostasis in aged hearts. Collectively, these findings establish cardiac lymphatic dysfunction and VEGFC deficiency as key features of cardiac aging, highlighting potential therapeutic entry points for age-associated heart disease.biostudies-arrayexpressSequencing - After library quality control by capillary electrophoresis (4200 TapeStation, Agilent), cDNA libraries were sequenced on the Illumina NextSeq 2000 platform generating 50 bp paired-end reads.Nucleic Acid Extraction - For genome-wide analysis of gene expression, RNA sequencing libraries from isolated mRNA were generated and sequenced by the Institute for Lung Health (ILH) – Genomics and Bioinformatics – at the Justus-Liebig-University (JLU) Giessen (Germany).Library Construction - A total amount of 400 ng of RNA per sample was used to enrich for polyadenylated mRNA followed by cDNA sequencing library preparation utilizing the Illumina® Stranded mRNA Prep Kit (Illumina) according to the manufacturer’s instructions.Sample Treatment - Lentiviral particles were generated by transfecting HEK293T cells (CRL-11268, ATCC) with the packing plasmids pCMV∆R8.91 and pMD2.G as well as the plasmids harboring the IL33 sequence (VB241001-1197hwt, VectorBuilder) or a stuffer sequence (VB241001-1200pax, VectorBuilder). IL33-overexpressing HDLECs were treated with recombinant VEGFC (200 ng/mL), Reelin (200 ng/mL), sphingosine-1-phosphate (200 ng/mL) for 24h.Sample Collection - Human dermal lymphatic endothelial cells (HDLECs) were purchased from PromoCell (C-12216) and cultured on fibronectin in PromoCell MV2 medium (C-22221) supplemented with the provided supplement mix (C-39221). Cells were cultured for 72h at 37°C and 5% CO2 at humidified atmosphere.Growth Protocol - HDLECs were treated with recombinant IFN-γ (30 ng/mL; 285-IF-100, R&D Systems), TNFα (10 ng/mL; 300-01A, Peprotech), IL1β (100 ng/mL; 201-LB-025, R&D Systems), G-CSF (100 ng/mL; AF-300-23-50UG, Gibco), GM-CSF (50 ng/mL; 300-03-20UG, Gibco), VEGFC (100 ng/mL; 9199-VC-025/CF, R&D Systems), Reelin (100 ng/mL; 8546-MR-050, R&D Systems), Apelin (100 ng/mL; ab152927, Abcam), sphingosine-1-phosphate (100 ng/mL; 73914-1MG, Merck) for 24h or with recombinant VEGFC (100 ng/mL), Reelin (100 ng/mL), Apelin (100 ng/mL), sphingosine-1-phosphate (100 ng/mL) for 48h. To assess autophagy, HDLECs were cultured in fully supplemented medium or Opti-MEM (51985026, Gibco) to starve the cells. Cells were treated with 10 µM chloroquine for 2h prior to fixation. For JAK inhibition, HDLECs were cultured with 5 µM ruxolitinib (83405S, CST) for 24h.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - The resulting raw reads were assessed for quality, adapter content and duplication rates with FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc). Reads were aligned versus the Ensembl human genome version hg38 (Ensembl release 101) using STAR 2.7.9a. Reads overlapping multiple genes or aligning to multiple regions were excluded. A combined raw count matrix was calculated, and batch corrected per data set using DESeq2.UnknownTranscriptomicsGenomicsProteomicsNextSeq 2000RNA-seq of coding RNAHomo sapiensDavid JohnfalseAge-Associated Loss of Lymphatic Vessels Promotes Cardiac Inflammation (Bulk-RNA-SEQ)Aging is a major risk factor for cardiovascular diseases, yet the contribution of the lymphatic vasculature to cardiac aging remains largely unexplored. Here, we show that aging reduces lymphatic vessel density in human and mice hearts and induces morphological changes, including the formation of zipper-like, tighter endothelial junctions. These alterations are accompanied by immune cell infiltration, fibrinogen and amyloid accumulation, and myocardial edema. Experimental reduction of cardiac lymphatics in young mice, achieved by Flt4 (VEGFR3) depletion or overexpression of soluble Flt4, reproduces some age-related cardiac phenotypes, such as inflammation and impaired lymphatic integrity. Mechanistically, we found that aging induces the selective up-regulation of nuclear interleukin 33 (IL33) in lymphatic endothelial cells. In contrast to the extracellular, cardioprotective form of IL33, nuclear IL33 promotes lymphatic cell death and junctional remodeling. A targeted screen of pro-lymphatic factors identified VEGFC as an age-sensitive regulator that both declines in the aging heart and suppresses IL33. Cardiac Vegfc overexpression or Il33 silencing restores lymphatic vessel density, reduces macrophage infiltration, and improves tissue homeostasis in aged hearts. Collectively, these findings establish cardiac lymphatic dysfunction and VEGFC deficiency as key features of cardiac aging, highlighting potential therapeutic entry points for age-associated heart disease.2026-08-03T00:00:00Z2026-08-03T14:10:50.773Z2026-08-03T14:10:19.396ZE-MTAB-17485ERP203237EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003816EFO_0003738EFO_0004184EFO_0003969