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Article ; Online: Effect of cytokine-induced alterations in extracellular matrix composition on diabetic retinopathy-relevant endothelial cell behaviors

Meredith J. Giblin / Cayla D. Ontko / John S. Penn

Scientific Reports, Vol 12, Iss 1, Pp 1-

2022 Volume 14

Abstract: Abstract Retinal vascular basement membrane (BM) thickening is an early structural abnormality of diabetic retinopathy (DR). Recent studies suggest that BM thickening contributes to the DR pathological cascade; however, much remains to be elucidated ... ...

Abstract	Abstract Retinal vascular basement membrane (BM) thickening is an early structural abnormality of diabetic retinopathy (DR). Recent studies suggest that BM thickening contributes to the DR pathological cascade; however, much remains to be elucidated about the exact mechanisms by which BM thickening develops and subsequently drives other pathogenic events in DR. Therefore, we undertook a systematic analysis to understand how human retinal microvascular endothelial cells (hRMEC) and human retinal pericytes (hRP) change their expression of key extracellular matrix (ECM) constituents when treated with diabetes-relevant stimuli designed to model the three major insults of the diabetic environment: hyperglycemia, dyslipidemia, and inflammation. TNFα and IL-1β caused the most potent and consistent changes in ECM expression in both hRMEC and hRP. We also demonstrate that conditioned media from IL-1β-treated human Müller cells caused dose-dependent, significant increases in collagen IV and agrin expression in hRMEC. After narrowing our focus to inflammation-induced changes, we sought to understand how ECM deposited by hRMEC and hRP under inflammatory conditions affects the behavior of naïve hRMEC. Our data demonstrated that diabetes-relevant alterations in ECM composition alone cause both increased adhesion molecule expression by and increased peripheral blood mononuclear cell (PBMC) adhesion to naïve hRMEC. Taken together, these data demonstrate novel roles for inflammation and pericytes in driving BM pathology and suggest that inflammation-induced ECM alterations may advance other pathogenic behaviors in DR, including leukostasis.
Keywords	Medicine ; R ; Science ; Q
Subject code	571
Language	English
Publishing date	2022-07-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Knowledge of an inflammatory biomarker of cardiovascular risk leads to biomarker-based decreased risk in pre-diabetic and diabetic patients

Diego Alcivar-Franco / Scott Purvis / Marc S. Penn / Andrea Klemes

Journal of International Medical Research, Vol

2020 Volume 48

Abstract: Objective Diabetes is a risk equivalent for cardiovascular events. The increase in vascular inflammation with diabetes is believed to be responsible for increased risk of ischemic events in diabetic patients. Our goal was to assess whether knowledge of ... ...

Abstract	Objective Diabetes is a risk equivalent for cardiovascular events. The increase in vascular inflammation with diabetes is believed to be responsible for increased risk of ischemic events in diabetic patients. Our goal was to assess whether knowledge of vascular inflammation alters cardiovascular risk over time, and how knowledge of vascular inflammation changes risk in non-diabetic, pre-diabetic and diabetic patients. Methods We retrospectively studied >100,000 primary-care patients per annum for 5 years (baseline in 2011 through 2015) with tests including lipoprotein profile, hemoglobin A1C and the vascular-specific inflammation risk marker myeloperoxidase. Results were obtained during the patient’s MD Value In Prevention (MDVIP) annual wellness program physical. Results We show that rates of patients with elevated myeloperoxidase levels were reduced from 14.4%, 15.2% and 21.3% to 4.0%, 4.0% and 6.7% in non-diabetic, pre-diabetic and diabetic patients, respectively, over the 5-year period. Decreases in vascular inflammation were achieved without decreases in the prevalence of pre-diabetes (hemoglobin A1C 5.7%–6.4%) or diabetes (hemoglobin A1C >6.4%) and were observed in patients below or above guideline low-density lipoprotein targets. Conclusions These data demonstrate that physicians informed of elevated markers of vascular inflammation can lower vascular inflammation correlating with biomarker-based decreased risk of cardiovascular events.
Keywords	Medicine (General) ; R5-920
Subject code	610
Language	English
Publishing date	2020-01-01T00:00:00Z
Publisher	SAGE Publishing
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Palmitic Acid Induces Müller Cell Inflammation that is Potentiated by Co-treatment with Glucose

Megan E. Capozzi / Meredith J. Giblin / John S. Penn

Scientific Reports, Vol 8, Iss 1, Pp 1-

2018 Volume 9

Abstract: Abstract Chronic hyperglycemia is thought to be the major stimulator of retinal dysfunction in diabetic retinopathy (DR). Thus, many diabetes-related systemic factors have been overlooked as inducers of DR pathology. Cell culture models of retinal cell ... ...

Abstract	Abstract Chronic hyperglycemia is thought to be the major stimulator of retinal dysfunction in diabetic retinopathy (DR). Thus, many diabetes-related systemic factors have been overlooked as inducers of DR pathology. Cell culture models of retinal cell types are frequently used to mechanistically study DR, but appropriate stimulators of DR-like factors are difficult to identify. Furthermore, elevated glucose, a gold standard for cell culture treatments, yields little to no response from many primary human retinal cells. Thus, the goal of this project was to demonstrate the effectiveness of the free fatty acid, palmitic acid and compare its use alone and in combination with elevated glucose as a stimulus for human Müller cells, a retinal glial cell type that is activated early in DR pathogenesis and uniquely responsive to fatty acids. Using RNA sequencing, we identified a variety of DR-relevant pathways, including NFκB signaling and inflammation, intracellular lipid signaling, angiogenesis, and MAPK signaling, that were stimulated by palmitic acid, while elevated glucose alone did not significantly alter any diabetes-relevant pathways. Co-treatment of high glucose with palmitic acid potentiated the expression of several DR-relevant angiogenic and inflammatory targets, including PTGS2 (COX-2) and CXCL8 (IL-8).
Keywords	Medicine ; R ; Science ; Q
Subject code	571
Language	English
Publishing date	2018-04-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Cytochrome P450-epoxygenated fatty acids inhibit Müller glial inflammation

Cayla D. Ontko / Megan E. Capozzi / Minjae J. Kim / Gary W. McCollum / John S. Penn

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 11

Abstract: Abstract Free fatty acid dysregulation in diabetics may elicit the release of inflammatory cytokines from Müller cells (MC), promoting the onset and progression of diabetic retinopathy (DR). Palmitic acid (PA) is elevated in the sera of diabetics and ... ...

Abstract	Abstract Free fatty acid dysregulation in diabetics may elicit the release of inflammatory cytokines from Müller cells (MC), promoting the onset and progression of diabetic retinopathy (DR). Palmitic acid (PA) is elevated in the sera of diabetics and stimulates the production of the DR-relevant cytokines by MC, including IL-1β, which induces the production of itself and other inflammatory cytokines in the retina as well. In this study we propose that experimental elevation of cytochrome P450 epoxygenase (CYP)-derived epoxygenated fatty acids, epoxyeicosatrienoic acid (EET) and epoxydocosapentaenoic acid (EDP), will reduce PA- and IL-1β-induced MC inflammation. Broad-spectrum CYP inhibition by SKF-525a increased MC expression of inflammatory cytokines. Exogenous 11,12-EET and 19,20-EDP significantly decreased PA- and IL-1β-induced MC expression of IL-1β and IL-6. Both epoxygenated fatty acids significantly decreased IL-8 expression in IL-1β-induced MC and TNFα in PA-induced MC. Interestingly, 11,12-EET and 19,20-EDP significantly increased TNFα in IL-1β-treated MC. GSK2256294, a soluble epoxide hydrolase (sEH) inhibitor, significantly reduced PA- and IL-1β-stimulated MC cytokine expression. 11,12-EET and 19,20-EDP were also found to decrease PA- and IL-1β-induced NFκB-dependent transcriptional activity. These data suggest that experimental elevation of 11,12-EET and 19,20-EDP decreases MC inflammation in part by blocking NFκB-dependent transcription and may represent a viable therapeutic strategy for inhibition of early retinal inflammation in DR.
Keywords	Medicine ; R ; Science ; Q
Subject code	571
Language	English
Publishing date	2021-05-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: A novel method for visualizing and tracking endogenous mRNA in a specific cell population in pathological neovascularization

Md Imam Uddin / Tyler C. Kilburn / Sara Z. Jamal / Craig L. Duvall / John S. Penn

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 16

Abstract: Abstract Diabetic retinopathy, retinopathy of prematurity and retinal vein occlusion are potentially blinding conditions largely due to their respective neovascular components. The development of real-time in vivo molecular imaging methods, to assess ... ...

Abstract	Abstract Diabetic retinopathy, retinopathy of prematurity and retinal vein occlusion are potentially blinding conditions largely due to their respective neovascular components. The development of real-time in vivo molecular imaging methods, to assess levels of retinal neovascularization (NV), would greatly benefit patients afflicted with these conditions. mRNA hybridization techniques offer a potential method to image retinal NV. The success of these techniques hinges on the selection of a target mRNA whose tissue levels and spatial expression patterns correlate closely with disease burden. Using a model of oxygen-induced retinopathy (OIR), we previously observed dramatic increases in retinal endoglin that localized to neovascular structures (NV), directly correlating with levels of neovascular pathology. Based on these findings, we have investigated Endoglin mRNA as a potential marker for imaging retinal NV in OIR mice. Also of critical importance, is the application of innovative technologies capable of detecting mRNAs in living systems with high sensitivity and specificity. To detect and visualize endoglin mRNA in OIR mice, we have designed and synthesized a novel imaging probe composed of short-hairpin anti-sense (AS) endoglin RNA coupled to a fluorophore and black hole quencher (AS-Eng shRNA). This assembly allows highly sensitive fluorescence emission upon hybridization of the AS-Eng shRNA to cellular endoglin mRNA. The AS-Eng shRNA is further conjugated to a diacyl-lipid (AS-Eng shRNA–lipid referred to as probe). The lipid moiety binds to serum albumin facilitating enhanced systemic circulation of the probe. OIR mice received intraperitoneal injections of AS-Eng shRNA–lipid. Ex vivo imaging of their retinas revealed specific endoglin mRNA dependent fluorescence superimposed on neovascular structures. Room air mice receiving AS-Eng shRNA–lipid and OIR mice receiving a non-sense control probe showed little fluorescence activity. In addition, we found that cells in neovascular lesions labelled with endoglin ...
Keywords	Medicine ; R ; Science ; Q
Subject code	500
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Author Correction

Cayla D. Ontko / Megan E. Capozzi / Minjae J. Kim / Gary W. McCollum / John S. Penn

Scientific Reports, Vol 11, Iss 1, Pp 1-

Cytochrome P450-epoxygenated fatty acids inhibit Müller glial inflammation

2021 Volume 1

Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2021-09-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Quartz Crystal Microbalance: Sensing Cell-Substrate Adhesion and Beyond

Chen, Jennifer Y / Lynn S. Penn / Jun Xi

Biosensors & bioelectronics. 2017,

2017

Abstract: Cell adhesion is an essential aspect of cellular behavior. Finding innovative methods to probe the adhesion of cells in their native state can greatly advance the understanding of control and regulation of cellular behavior and their impact on human ... ...

Abstract	Cell adhesion is an essential aspect of cellular behavior. Finding innovative methods to probe the adhesion of cells in their native state can greatly advance the understanding of control and regulation of cellular behavior and their impact on human health. The quartz crystal microbalance (QCM) is a label-free, biosensing system that has, in the past fifty years, evolved from a simple acoustic based mass sensor to a powerful bioanalytical tool. Its unique capability of monitoring the cell-substrate interaction non-invasively in real time has led to the emergence of its applications in areas that are relevant to fundamental cell biology and medical research. This review is intended to provide readers an overview of the use of the QCM for examination of cell-substrate adhesion. It also describes how this innovative approach can be extended to the study of other aspects of cellular behavior, such as cell morphology, cell mechanics, cell motility, cell signaling, all of which can potentially be applied to medical diagnosis and/or pharmaceutical development. In this review a major emphasis is placed on informing readers about some of the most important practical aspects of the QCM-based cell study including data acquisition and analysis, the substrate surface manipulation, and cell manipulation.
Keywords	acoustics ; biomedical research ; biosensors ; cell adhesion ; cell movement ; human health ; mechanics ; monitoring ; quartz
Language	English
Size	p. .
Publishing place	Elsevier B.V.
Document type	Article
Note	Pre-press version
ZDB-ID	1011023-9
ISSN	1873-4235 ; 0956-5663
ISSN (online)	1873-4235
ISSN	0956-5663
DOI	10.1016/j.bios.2017.08.032
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: RNA-Seq reveals a role for NFAT-signaling in human retinal microvascular endothelial cells treated with TNFα.

Sara R Savage / Colin A Bretz / John S Penn

PLoS ONE, Vol 10, Iss 1, p e

2015 Volume 0116941

Abstract: TNFα has been identified as playing an important role in pathologic complications associated with diabetic retinopathy and retinal inflammation, such as retinal leukostasis. However, the transcriptional effects of TNFα on retinal microvascular ... ...

Abstract	TNFα has been identified as playing an important role in pathologic complications associated with diabetic retinopathy and retinal inflammation, such as retinal leukostasis. However, the transcriptional effects of TNFα on retinal microvascular endothelial cells and the different signaling pathways involved are not yet fully understood. In the present study, RNA-seq was used to profile the transcriptome of human retinal microvascular endothelial cells (HRMEC) treated for 4 hours with TNFα in the presence or absence of the NFAT-specific inhibitor INCA-6, in order to gain insight into the specific effects of TNFα on RMEC and identify any involvement of NFAT signaling. Differential expression analysis revealed that TNFα treatment significantly upregulated the expression of 579 genes when compared to vehicle-treated controls, and subsequent pathway analysis revealed a TNFα-induced enrichment of transcripts associated with cytokine-cytokine receptor interactions, cell adhesion molecules, and leukocyte transendothelial migration. Differential expression analysis comparing TNFα-treated cells to those co-treated with INCA-6 revealed 10 genes whose expression was significantly reduced by the NFAT inhibitor, including those encoding the proteins VCAM1 and CX3CL1 and cytokines CXCL10 and CXCL11. This study identifies the transcriptional effects of TNFα on HRMEC, highlighting its involvement in multiple pathways that contribute to retinal leukostasis, and identifying a previously unknown role for NFAT-signaling downstream of TNFα.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2015-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: In vivo photothermal optical coherence tomography of endogenous and exogenous contrast agents in the eye

Maryse Lapierre-Landry / Andrew Y. Gordon / John S. Penn / Melissa C. Skala

Scientific Reports, Vol 7, Iss 1, Pp 1-

2017 Volume 9

Abstract: Abstract Optical coherence tomography (OCT) has become a standard-of-care in retinal imaging. OCT allows non-invasive imaging of the tissue structure but lacks specificity to contrast agents that could be used for in vivo molecular imaging. Photothermal ... ...

Abstract	Abstract Optical coherence tomography (OCT) has become a standard-of-care in retinal imaging. OCT allows non-invasive imaging of the tissue structure but lacks specificity to contrast agents that could be used for in vivo molecular imaging. Photothermal OCT (PT-OCT) is a functional OCT-based technique that has been developed to detect absorbers in a sample. We demonstrate in vivo PT-OCT in the eye for the first time on both endogenous (melanin) and exogenous (gold nanorods) absorbers. Pigmented mice and albino mice (n = 6 eyes) were used to isolate the photothermal signal from the melanin in the retina. Pigmented mice with laser-induced choroidal neovascularization lesions (n = 7 eyes) were also imaged after a systemic injection of gold nanorods to observe their passive accumulation in the retina. This experiment demonstrates the feasibility of PT-OCT to image the distribution of both endogenous and exogenous absorbers in the mouse retina.
Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2017-08-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Synthesis of macromolecular mimics of small leucine-rich proteoglycans with a poly(ethylene glycol) core and chondroitin sulphate bristles

Sarkar, Sumona / Carli Moorehead / Caroline L. Schauer / Katsiaryna Prudnikova / Lynn S. Penn / Michele Marcolongo

Carbohydrate polymers. 2017 June 15, v. 166

2017

Abstract: Small leucine-rich proteoglycans (SLRPs) are a class of molecules prevalent in almost all tissues types and are thought to be responsible for collagen organization and macro-scale biological properties. However, when they are dysfunctional or degraded, ... ...

Abstract	Small leucine-rich proteoglycans (SLRPs) are a class of molecules prevalent in almost all tissues types and are thought to be responsible for collagen organization and macro-scale biological properties. However, when they are dysfunctional or degraded, severe pathological phenotypes are observed. Here we investigate macromolecular mimics to SLRPs using poly(ethylene glycol) (PEG) as a core (replacing the protein core of natural SLRPs) and chondroitin sulphate (CS) bristle(s) in an end-on attachment (via epoxide-amine reactions), mimicking the physical structure of the natural SLRPs. Poly(ethylene glycol)-diglycidyl ether (PEG-DEG) and ethylene glycol-diglycidyl ether (EG-DGE) monomers were used to incorporate CS bristles into a macromolecule that closely mimics the SLRP biglycan structure in a grafting-to strategy. The kinetics of these reactions was studied along with the specific viscosity and cytocompatibility of resulting CS macromolecules. Structures were found to incorporate two CS chains (similar to biglycan) on average and exhibited cytocompatibility equivalent to or better than CS-only controls.
Keywords	chondroitin sulfate ; collagen ; ethylene glycol ; phenotype ; polyethylene glycol ; proteoglycans ; setae (animal) ; tissues ; viscosity
Language	English
Dates of publication	2017-0615
Size	p. 338-347.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	1501516-6
ISSN	1879-1344 ; 0144-8617
ISSN (online)	1879-1344
ISSN	0144-8617
DOI	10.1016/j.carbpol.2017.02.083
Database	NAL-Catalogue (AGRICOLA)

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