Analysis of myocardial cellular gene expression during pressure overload reveals matrix based functional intercellular communication

Froese, Natali; Cordero, Julio; Abouissa, Aya; Trogisch, Felix A.; Grein, Steve; Szaroszyk, Malgorzata; Wang, Yong; Gigina, Anna; Korf‐Klingebiel, Mortimer; Bošnjak, Berislav; Davenport, Colin; Wiehlmann, Lutz; Geffers, Robert; Riechert, Eva; Jürgensen, Lonny; Boileau, Etienne; Lin, Yanzhu; Dieterich, Christoph; Förster, Reinhold; Bauersachs, Johann; Ola, Roxana; Dobreva, Gergana; Völkers, Mirko; Heineke, Joerg

doi:10.1016/j.isci.2022.103965

Cited by 16 publications

(27 citation statements)

References 71 publications

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“…The purity of EC in our MACS based isolation procedure is >90%. 30 The genome-wide transcriptome analysis via bulk RNAseq identified a total of 3355 differentially expressed (DE) genes between WT and Gadlor- KO, which are shown in the heatmap (Supplement Figure 6A) . Gene ontology analysis from DE genes and RT-qPCR revealed and confirmed an upregulation of angiogenesis, mitotic cell-cycle and respiratory electron transport and a downregulation of inflammatory response genes in Gadlor -KO ECs after 2-weeks TAC (Supplement Figure 6B-D) .…”

Section: Resultsmentioning

confidence: 99%

“…The purity of cardiac FBs in our MACS-based isolation procedure is >90%. 34 Genome-wide analysis revealed a total of 915 downregulated and 998 upregulated genes in Gadlor- KO FBs compared to WT mice after TAC, which are shown in a heatmap in Figure S5A . Amongst these, we detected a strong downregulation of mainly ECM organization and growth factor associated genes (confirmed by RT-qPCR: Fgfr1, Igf1, Col1a1, Col3a1, Col6a1 ) (Figure S5B-D) .…”

Section: Resultsmentioning

confidence: 99%

“…The purity of cardiac FBs in our MACS based isolation procedure is >90%. 27 Genome-wide analysis revealed total of 915 downregulated and 998 upregulated genes in Gadlor-KO FBs compared to WT mice after TAC, which are shown in a heatmap in Figure 5A.Amongst these, we detected a strong downregulation of mainly ECM organization associated genes (confirmed by Igf1, Col1a1, Col3a1, Col6a1) (Figure 5B-D). Additionally, genes related to the regulation of angiogenesis, blood vessel and endothelium development were upregulated in Gadlor-KO FBs, which suggested a contribution of FBs for vessel development and angiogenesis observed in Gadlor-KO ECs upon overload.…”

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confidence: 92%

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Endothelial cell-derived, secreted long non-coding RNAsGadlor1andGadlor2aggravate pathological cardiac remodeling via intercellular crosstalk

Keles

Grein

Froese

et al. 2022

Preprint

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Background: Pathological cardiac overload triggers maladaptive myocardial remodeling that predisposes to the development of heart failure. The contribution of long non-coding RNAs (lncRNAs) to intercellular signaling during cardiac remodeling is largely unknown. Methods: We analyzed the expression of Gadlor 1 and Gadlor2 lncRNAs in mouse hearts, mouse cardiac cells, extracellular vesicles (EVs), human failing hearts as well as patient serum. Gadlor knock-out (KO) mice were generated and analyzed. The effect of Gadlor knock-out and Gadlor overexpression during cardiac pressure overload induced by transverse aortic constriction (TAC) was analyzed by echocardiography, histological analyses and RNA sequencing in isolated cardiac cells. Gadlor1/2 interaction partners were identified by RNA antisense purification coupled with mass-spectrometry (RAP-MS). Results: In the heart, the related lncRNAs Gadlor1 and 2 are mainly expressed in endothelial cells and to a lesser extent in fibroblasts. Gadlor1/2 are upregulated in failing mouse hearts as well as in the myocardium and in serum of heart failure patients. Interestingly, Gadlor1 and 2 are secreted from endothelial cells within EVs, which are taken up by cardiomyocytes. Gadlor-KO mice exerted reduced cardiomyocyte hypertrophy, diminished myocardial fibrosis and improved cardiac function, but paradoxically suffered from sudden death during prolonged overload. Gadlor overexpression, in turn, triggered hypertrophy, fibrosis and cardiac dysfunction. Mechanistically, Gadlor1 and Gadlor2 inhibit angiogenic gene expression in endothelial cells, while promoting the expression of pro-fibrotic genes in cardiac fibroblasts. In cardiomyocytes, Gadlor1/2 upregulate mitochondrial genes, but downregulate angiogenesis genes, while interacting with the transcriptional regulator Glyr1 and the calcium/calmodulin-dependent protein kinase type II (CaMKII), entailing cardiomyocyte hypertrophy and perturbed cardiomyocyte calcium dynamics. Conclusions: We describe that Gadlor1 and 2, two related, novel lncRNAs, are upregulated in cardiac pathological overload and are secreted from endothelial cells within EVs. Gadlor1/2 induce cardiac dysfunction, cardiomyocyte hypertrophy and myocardial fibrosis by exerting heterocellular effects on cardiac cellular gene-expression and by affecting calcium dynamics in cardiomyocytes, which take up the Gadlor1/2 by EV mediated transfer from endothelial cells. Targeted inhibition of Gadlor lncRNAs in endothelial cells or fibroblasts might serve as a therapeutic strategy in the future.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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confidence: 92%

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Endothelial cell-derived, secreted long non-coding RNAsGadlor1andGadlor2aggravate pathological cardiac remodeling via intercellular crosstalk

Keles

Grein

Froese

et al. 2022

Preprint

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“…Adult heart endothelial cells were isolated from the hearts of adult mice using CD146-coated micro beads with MACS technology from Miltenyi. The primary isolated ECs were >90% pure, as determined by FACS analysis after staining for CD31 [ 21 ]. The isolated cells were directly used for RNA extraction.…”

Section: Methodsmentioning

confidence: 99%

“…All cells were harvested 48 h after transfection/infection using Passive Lysis Buffer (Promega, Madison, WI, USA) and luciferase activity was measured. Values were normalized to total protein as described previously [ 21 ].…”

Section: Methodsmentioning

confidence: 99%

Endothelial Cell GATA2 Modulates the Cardiomyocyte Stress Response through the Regulation of Two Long Non-Coding RNAs

Froese

Szaroszyk

Korf‐Klingebiel

et al. 2022

Biology

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Capillary endothelial cells modulate myocardial growth and function during pathological stress, but it is unknown how and whether this contributes to the development of heart failure. We found that the endothelial cell transcription factor GATA2 is downregulated in human failing myocardium. Endothelial GATA2 knock-out (G2-EC-KO) mice develop heart failure and defective myocardial signal transduction during pressure overload, indicating that the GATA2 downregulation is maladaptive. Heart failure and perturbed signaling in G2-EC-KO mice could be induced by strong upregulation of two unknown, endothelial cell-derived long non-coding (lnc) RNAs (AK037972, AK038629, termed here GADLOR1 and 2). Mechanistically, the GADLOR1/2 lncRNAs transfer from endothelial cells to cardiomyocytes, where they block stress-induced signalling. Thereby, lncRNAs can contribute to disease as paracrine effectors of signal transduction and therefore might serve as therapeutic targets in the future.

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Inter- and Intracellular Signaling Pathways

Dobreva,

Heineke

2024

Advances in Experimental Medicine and Biology

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Analysis of myocardial cellular gene expression during pressure overload reveals matrix based functional intercellular communication

Cited by 16 publications

References 71 publications

Endothelial cell-derived, secreted long non-coding RNAsGadlor1andGadlor2aggravate pathological cardiac remodeling via intercellular crosstalk

Endothelial cell-derived, secreted long non-coding RNAsGadlor1andGadlor2aggravate pathological cardiac remodeling via intercellular crosstalk

Endothelial Cell GATA2 Modulates the Cardiomyocyte Stress Response through the Regulation of Two Long Non-Coding RNAs

Inter- and Intracellular Signaling Pathways

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