Rap1a Regulates Cardiac Fibroblast Contraction of 3D Diabetic Collagen Matrices by Increased Activation of the AGE/RAGE Cascade

Abstract

Cardiovascular disease is a common diabetic complication that can arise when cardiac fibroblasts transition into myofibroblasts. Myofibroblast transition can be induced by advanced glycated end products (AGEs) present in the extracellular matrix (ECM) activating RAGE (receptor for advanced glycated end products) to elicit intracellular signaling. The levels of AGEs are higher under diabetic conditions due to the hyperglycemic conditions present in diabetics. AGE/RAGE signaling has been shown to alter protein expression and ROS production in cardiac fibroblasts, resulting in changes in cellular function, such as migration and contraction. Recently, a small GTPase, Rap1a, has been identified to overlap the AGE/RAGE signaling cascade and mediate changes in protein expression. While Rap1a has been shown to impact AGE/RAGE-induced protein expression, there are currently no data examining the impact Rap1a has on AGE/RAGE-induced cardiac fibroblast function. Therefore, we aimed to determine the impact of Rap1a on AGE/RAGE-mediated cardiac fibroblast contraction, as well as the influence isolated diabetic ECM has on facilitating these effects. In order to address this idea, genetically different cardiac fibroblasts were embedded in 3D collagen matrices consisting of collagen isolated from either non-diabetic of diabetic mice. Fibroblasts were treated with EPAC and/or exogenous AGEs, which was followed by assessment of matrix contraction, protein expression (α-SMA, SOD-1, and SOD-2), and hydrogen peroxide production. The results showed Rap1a overlaps the AGE/RAGE cascade to increase the myofibroblast population and generation of ROS production. The increase in myofibroblasts and oxidative stress appeared to contribute to increased matrix contraction, which was further exacerbated by diabetic conditions. Based off these results, we determined that Rap1a was essential in mediating the response of cardiac fibroblasts to AGEs within diabetic collagen.

Age-Related Alterations of Hyaluronan and Collagen in Extracellular Matrix of the Muscle Spindles

“…These results have confirmed the findings of another study demonstrating that older adults have thicker MS capsules [ 11 ]. Moreover, these findings are consistent with those of some studies that highlighted the role of the collagens in various diseases and tissues [ 21 , 22 , 23 ]. Parkes et al [ 21 ] reported the prominent role of collagen and HA in the ECM of ovaries, suggesting their crucial activity for ovarian homeostasis, possibly through signaling events or tissue micromechanism.…”

“…Numerous reports described the association between the single-nucleotide polymorphism rs12722 and rs13936 in the COL5A1 gene and injuries, such as Achilles tendon pathology, anterior cruciate ligament, and tennis elbow [ 41 , 42 , 43 , 44 , 45 ]. Daleswski et al [ 22 ] studied the COL5A1 rs12722 and rs13946 polymorphisms as potential genetic factors regulating the ADDwoR-mediated soft tissue pathway in association with temporomandibular joint anterior disc displacement [ 22 ]. Pirri et al [ 46 ] reported that:” excessive aggregation of ECM elements is present in fibrosis (e.g., in myopathies), including in fasciae, and occurs during aging [ 16 ], and diabetes, characterized by increased endomysium as well as perimysium” [ 47 ].…”

“…Cell behavior is profoundly affected by the environment in which they live, thus the aging of the ECM is central to understanding age-related changes [ 17 ]. Nowadays, ECM’s role has been highlighted in different diseases and tissues [ 19 , 20 , 21 , 22 , 23 ].…”

Exploring the mechanism by which aqueous Gynura divaricata inhibits diabetic foot based on network pharmacology, molecular docking and experimental verification

“…The KEGG results suggest that the AGE-RAGE signaling pathway is likely the most relevant pathway for GD. Studies have demonstrated that the specific combination of AGE and RAGE is an important mechanism that affects the level of oxidative stress in diabetic wounds (Burr et al 2021a , b ). The AGE-RAGE signaling pathway is a principal pathway in the pathogenesis of inflammatory diseases.…”

Integrated proteomics and metabolomics analysis of cardiac-tissue in mice with sepsis

“…Rap1a, a GTPase, connects external stimuli to intracellular responses by inducing changes in effector proteins (Wang et al, 2015;Yan et al, 2008). Research has shown that cardiac broblasts' response is mediated by Rap1a (Burr & Stewart, 2021). There are four distinct isoforms of phospholipase C-β3 (PLC-β3), which are PLC-β1, 2, 3, and 4, with PLC-β1 being the most abundant.…”