AKAP12 deficiency impairs VEGF‐induced endothelial cell migration and sprouting

Benz, P.; Ding, Yindi; Stingl, Heike; Loot, Annemarieke E.; Zink, Joana; Wittig, Ilka; Popp, Rüdiger; Fleming, Ingrid

doi:10.1111/apha.13325

Cited by 38 publications

(35 citation statements)

References 70 publications

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“…CHD and specifically right ventricular outlet obstruction are present in a fraction of the cases (39). The results also suggested other constrained genes not previously identified, with evidence that supports a role in the VEGF pathway or other complementary mechanisms for TOF, from human (e.g., AKAP12 ), mouse (e.g., EPN1 , ATF2 ) or both ( PKD1 ) (Table 3) derived gene-sets (40–44).…”

Section: Discussionsupporting

confidence: 68%

Genes and pathways implicated in tetralogy of Fallot revealed by ultra-rare variant burden analysis in 231 genome sequences

Manshaei

Merico

Reuter

et al. 2020

Preprint

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Recent genome-wide studies of rare genetic variants have begun to implicate novel mechanisms for tetralogy of Fallot (TOF), a severe congenital heart defect (CHD).To provide statistical support for case-only data without parental genomes, we re-analyzed genome sequences of 231 individuals with TOF or related CHD. We adapted a burden test originally developed for de novo variants to assess singleton variant burden in individual genes, and in gene-sets corresponding to functional pathways and mouse phenotypes, accounting for highly correlated gene-sets, and for multiple testing.The gene burden test identified a significant burden of deleterious missense variants in NOTCH1 (Bonferroni-corrected p-value <0.01). These NOTCH1 variants showed significant enrichment for those affecting the extracellular domain, and especially for disruption of cysteine residues forming disulfide bonds (OR 39.8 vs gnomAD). Individuals with NOTCH1 variants, all with TOF, were enriched for positive family history of CHD. Other genes not previously implicated in TOF had more modest statistical support and singleton missense variant results were non-significant for gene-set burden. For singleton truncating variants, the gene burden test confirmed significant burden in FLT4. Gene-set burden tests identified a cluster of pathways corresponding to VEGF signaling (FDR=0%), and of mouse phenotypes corresponding to abnormal vasculature (FDR=0.8%), that suggested additional candidate genes not previously identified (e.g., WNT5A and ZFAND5). Analyses using unrelated sequencing datasets supported specificity of the findings for CHD.The findings support the importance of ultra-rare variants disrupting genes involved in VEGF and NOTCH signaling in the genetic architecture of TOF. These proof-of-principle data indicate that this statistical methodology could assist in analyzing case-only sequencing data in which ultra-rare variants, whether de novo or inherited, contribute to the genetic etiopathogenesis of a complex disorder.Author summaryWe analyzed the ultra-rare nonsynonymous variant burden for genome sequencing data from 231 individuals with congenital heart defects, most with tetralogy of Fallot. We adapted a burden test originally developed for de novo variants. In line with other studies, we identified a significant truncating variant burden for FLT4 and deleterious missense burden for NOTCH1, both passing a stringent Bonferroni multiple-test correction. For NOTCH1, we observed frequent disruption of cysteine residues establishing disulfide bonds in the extracellular domain. We also identified genes with BH-FDR <10% that were not previously implicated. To overcome limited power for individual genes, we tested gene-sets corresponding to functional pathways and mouse phenotypes. Gene-set burden of truncating variants was significant for vascular endothelial growth factor signaling and abnormal vasculature phenotypes. These results confirmed previous findings and suggested additional candidate genes for experimental validation in future studies. This methodology can be extended to other case-only sequencing data in which ultra-rare variants make a substantial contribution to genetic etiology.

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Section: Discussionsupporting

confidence: 68%

Genes and pathways implicated in tetralogy of Fallot revealed by ultra-rare variant burden analysis in 231 genome sequences

Manshaei

Merico

Reuter

et al. 2020

Preprint

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“…For instance, studies have shown that the pan-cellular increases in cAMP caused by agents that activate all transmembrane adenylyl cyclases, including agents such as forskolin, or which inhibit all cellular cAMP-hydrolyzing phosphodiesterases (PDEs), like isobutyl-methyl-xanthine (IBMX), consistently reduce ASMC migration [1]. For these reasons, cAMP-elevating agents have long been seen as attractive agents through which to reduce ASMC migration in several conditions, including in-stent restenosis, where PDE4 inhibition reduces neointima formation and inhibits vascular cell adhesion molecule 1 (VCAM-1) expression and histone methylation in an exchange protein activated by cAMP (EPAC)-dependent manner [2,3] Interestingly, notwithstanding the observation that increased cAMP signaling results in reduced levels of ASMC migration, when the numerous steps involved in coordinating cellular migration are studied individually, it is found that they are equally likely to be inhibited or promoted [1,[4][5][6]. For instance, while increased protein kinase A (PKA) activity reduces overall cellular migration, PKA activation promotes, rather than inhibits, the formation of cellular protrusions at the cellular front, a critical early step in polarizing cells for migration [5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

An EPAC1/PDE1C-Signaling Axis Regulates Formation of Leading-Edge Protrusion in Polarized Human Arterial Vascular Smooth Muscle Cells

Brzezińska

Maurice

2019

Cells

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Pharmacological activation of protein kinase A (PKA) reduces migration of arterial smooth muscle cells (ASMCs), including those isolated from human arteries (HASMCs). However, when individual migration-associated cellular events, including the polarization of cells in the direction of movement or rearrangements of the actin cytoskeleton, are studied in isolation, these individual events can be either promoted or inhibited in response to PKA activation. While pharmacological inhibition or deficiency of exchange protein activated by cAMP-1 (EPAC1) reduces the overall migration of ASMCs, the impact of EPAC1 inhibition or deficiency, or of its activation, on individual migration-related events has not been investigated. Herein, we report that EPAC1 facilitates the formation of leading-edge protrusions (LEPs) in HASMCs, a critical early event in the cell polarization that underpins their migration. Thus, RNAi-mediated silencing, or the selective pharmacological inhibition, of EPAC1 decreased the formation of LEPs by these cells. Furthermore, we show that the ability of EPAC1 to promote LEP formation by migrating HASMCs is regulated by a phosphodiesterase 1C (PDE1C)-regulated “pool” of intracellular HASMC cAMP but not by those regulated by the more abundant PDE3 or PDE4 activities. Overall, our data are consistent with a role for EPAC1 in regulating the formation of LEPs by polarized HASMCs and show that PDE1C-mediated cAMP hydrolysis controls this localized event.

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“…In this issue of Acta Physiologica, Benz et al study the role of one important protein in the cyclic AMP signalling pathway, the A-kinase anchoring (AKAP)12. 1 The downstream effects stemming from cAMP release are tightly controlled and activate a profusion of signalling pathways. However, many of these different processes function with largely the same major constituent proteins, including adenylate cyclases, kinases, phosphatases and phosphodiesterases.…”

Section: E D I T O R I a L The Role Of Akap12 In Coordination Of Vegfmentioning

confidence: 99%

“…In this issue of Acta Physiologica, Benz and colleagues present their findings on the role AKAP12 plays in endothelial cell migration and angiogenesis. 1 Through a combination of biochemical, proteomic and imaging techniques, the authors detail how AKAP12 expression in migrating endothelial cells may coordinate the integration of angiogenic stimuli and PKA activity-induced changes in actin dynamics that drive cell movement. In this work, Benz and co-workers addressed both the differential expression of AKAP12 in migrating cells, and its functional role in coordinating signal integration that helps to drive endothelial cell motility.…”

Section: E D I T O R I a L The Role Of Akap12 In Coordination Of Vegfmentioning

confidence: 99%

“…1 Through a combination of biochemical, proteomic and imaging techniques, the authors detail how AKAP12 expression in migrating endothelial cells may coordinate the integration of angiogenic stimuli and PKA activity-induced changes in actin dynamics that drive cell movement. 1 Through a combination of biochemical, proteomic and imaging techniques, the authors detail how AKAP12 expression in migrating endothelial cells may coordinate the integration of angiogenic stimuli and PKA activity-induced changes in actin dynamics that drive cell movement.…”

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