<i>JNK2</i>Gene Silencing for Elastic Matrix Regenerative Repair

Carney, Sarah; Broekelmann, Thomas J.; Mecham, Robert P.; Ramamurthi, Anand

doi:10.1089/ten.tea.2020.0221

Cited by 2 publications

(10 citation statements)

References 48 publications

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“…Regardless of the ether and CH stretching peaks on PEGylated LNPs, both LNPs (with or without PEG) exhibited vibration peaks at 1536 cm −1 , and 1460 cm −1 corresponding to CC stretching and CN stretching (amide bonds), whereas peak at 1635 cm −1 represents esters, all of which are the bonds present in the lipid mixtures (DOTAP, DOPE, and Cholesterol). Our results correspond to other published findings 4,10 …”

Section: Discussionsupporting

confidence: 94%

“…This suggests that the knockdown of JNK2 might take between 24 h and 5 days to significantly attenuate their gene and protein expressions post LNP treatment. Although our siRNA is targeted against JNK2 , the decrease in MMP2 expression is a downstream effect of JNK2 attenuation, which aligns well with our prior published findings 4 . At day 5, JNK2 gene (Figure 7E) and protein (Figure 8A) expression were significantly reduced upon siRNA treatment versus the untreated control.…”

Section: Discussionsupporting

confidence: 91%

“…In addition, the increased intensity of the peak at 2915 and 2852 cm À1 on the PEGylated LNPs compared to the non-PEGylated LNPs were due to the extra CH stretching offered by the DSPE component which was absent in LNPs PEG.Regardless of the ether and CH stretching peaks on PEGylated LNPs, both LNPs (with or without PEG) exhibited vibration peaks at 1536 cm À1 , and 1460 cm À1 corresponding to C C stretching and CN stretching (amide bonds), whereas peak at 1635 cm À1 represents esters, all of which are the bonds present in the lipid mixtures (DOTAP, DOPE, and Cholesterol). Our results correspond to other published findings 4,10.…”

supporting

confidence: 94%

“…Previous studies have shown JNK2 to be a primary mediator of elastic matrix remodeling. 4,32 Attenuation or deletion of JNK2 gene leads to upregulated elastogenesis. 1,33 JNK-mediated improvement in elastogenesis occurs by (a) reduction in proteolytic activity of MMP2 resulting in a net decrease of elastic matrix degradation, 34 and (b) upregulation of elastic fiber assembly genes and proteins such as ELN, LOX, fibulins, and fibrillins.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies have shown JNK2 to be a primary mediator of elastic matrix remodeling 4,32 . Attenuation or deletion of JNK2 gene leads to upregulated elastogenesis 1,33 .…”

Section: Discussionmentioning

confidence: 99%

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JNK2 silencing lipid nanoparticles for elastic matrix repair

Dahal,

Bastola,

Ramamurthi

2023

J Biomedical Materials Res

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The over‐expression of c‐Jun N‐terminal kinase (JNK2), a stress activated mitogen kinase, in the aortic wall plays a critical role in the formation and progression of abdominal aortic aneurysm (AAA). This triggers chronic downstream upregulation of elastolytic matrix metalloproteinases (MMPs), MMPs2 and 9 to cause progressive proteolytic breakdown of the wall elastic matrix. We have previously shown that siNRA knockdown of JNK2 gene expression in an AAA culture model stimulates downstream elastin gene expression, elastic fiber formation, crosslinking and reduces elastolytic MMPs2 and 9. Since naked siRNA poorly routes to intracellular targets, has poor stability in blood, and could be potentially toxic and immunogenic, this project is aimed to develop PEGylated lipid nanoparticles (LNPs) for delivery of JNK siRNA and to generate evidence of successful JNK2 knockdown and downstream attenuation of MMP2 gene and protein expressions. LNPs were formulated using thin‐film hydration technique and had the size of 100–200 nm with zeta‐potential ranging between 30 and 40 mV. JNK siRNA loaded PEGylated LNPs successfully knocked down JNK2 in cytokine‐activated rat aneurysmal smooth muscle (EaRASMC) cultures. This resulted in a downstream decrease in MMP2 gene and protein expression and an upward trend in expression of genes for proteins critical for elastic fiber assembly such as elastin (ELN) and lysyl oxidase (LOX). Our result indicates cationic LNPs to be potential carriers for JNK siRNA delivery improving potency for elastin homeostasis required for AAA repair which could possibly provide benefits in preventing the progression of small AAAs.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionsupporting

confidence: 91%

supporting

confidence: 94%

Section: Discussionmentioning

confidence: 99%

“…Previous studies have shown JNK2 to be a primary mediator of elastic matrix remodeling 4,32 . Attenuation or deletion of JNK2 gene leads to upregulated elastogenesis 1,33 .…”

Section: Discussionmentioning

confidence: 99%

See 3 more Smart Citations

JNK2 silencing lipid nanoparticles for elastic matrix repair

Dahal,

Bastola,

Ramamurthi

2023

J Biomedical Materials Res

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Assessing trans‐endothelial transport of nanoparticles for delivery to abdominal aortic aneurysms

Yau,

Chukwu,

Jedlicka

et al. 2024

J Biomedical Materials Res

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Abdominal aortic aneurysms (AAAs) are localized, rupture‐prone expansions of the abdominal aorta wall. In this condition, structural extracellular matrix (ECM) proteins of the aorta wall, elastic fibers and collagen fibers, that impart elasticity and stiffness respectively, are slowly degraded by overexpressed matrix metalloproteinases (MMPs) following an injury stimulus. We are seeking to deliver therapeutics to the AAA wall using polymer nanoparticles (NPs) that are capable of stimulating on‐site matrix regeneration and repair. This study aimed to determine how NP shape and size impacts endocytosis and transmigration past the endothelial cell (EC) layer from circulation into the medial layer of the AAA wall. First, rod‐shaped NPs were shown to be created based mechanical stretching of PLGA NPs while embedded in a PVA film with longer rod‐shaped NPs created based of the degree in which the PVA films are stretched. Live/dead assay reveals that our PLGA NPs are safe and do not cause cell death. Immunofluorescence staining reveal cytokine activation causes endothelial dysfunction in ECs by increasing expression of inflammatory marker Integrin αVβ3 and decreasing expression of adhesion protein vascular endothelial (VE)‐cadherin. We showed this disruption enable greater EC uptake and translocation of NPs. Fluorescence studies demonstrate high endothelial transmigration and endocytosis with rod‐shaped NPs in cytokine activated ECs compared to healthy control cells, arguing for the benefits of using higher aspect ratio (AR) NPs for accumulation at the aneurysm site. We also demonstrated that the mechanisms of NP transmigration across an activated EC layer depend on NP AR. These results show the potential of using shape as a modality for enhancing permeation of NPs into the aneurysm wall. These studies are also significance to understanding the mechanisms that are likely engaged by NPs for penetrating the endothelial lining of aneurysmal wall segments.

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JNK2Gene Silencing for Elastic Matrix Regenerative Repair

Cited by 2 publications

References 48 publications

JNK2 silencing lipid nanoparticles for elastic matrix repair

JNK2 silencing lipid nanoparticles for elastic matrix repair

Assessing trans‐endothelial transport of nanoparticles for delivery to abdominal aortic aneurysms

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