Adenovirus-mediated gene transfer of hIGF-IB in mouse lungs induced prolonged inflammation but no fibroproliferation

Pulmonary fibrosis (PF), the end stage of a variety of fibroproliferative lung diseases, is characterized by excessive lung mesenchymal cell activation and extracellular matrix deposition. Most PF is induced after repetitive or chronic lung inflammation; however, a significant portion of PF occurs without apparent inflammation. The mechanisms of fibroproliferation are poorly understood. Studies have shown that cytokines regulating inflammation and tissue repair processes play essential roles in the development of PF. Insulin-like growth factor I (IGF-I) has been shown to stimulate lung mesenchymal cell proliferation and extracellular matrix synthesis in vitro and is significantly elevated in patients with PF. In this study, we investigated whether human IGF-IB (hIGF-IB) expression in the lungs induces PF in a C57BL/6 mouse model. Mice were subjected to adenoviral gene transfer, and the effects of hIGF-IB expression on the lungs were examined 3, 7, 14, 21, and 42 days after gene delivery. hIGF-IB expression induced significant and prolonged inflammatory cell infiltration into the lungs, with an early neutrophil infiltration followed by a late macrophage infiltration. No significant fibroblast or matrix accumulation could be detected in the lungs of these mice. No significant collagen accumulation could be detected in vivo, despite in vitro evidence that hIGF-IB induces collagen mRNA expression in fibroblasts. Therefore, IGF-IB alone is not sufficient to induce fibrosis, and it is possible that a coactivator is required to induce significant fibroproliferation in vivo.

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Formulation and in vitro and in vivo evaluation of a cationic emulsion as a vehicle for improving adenoviral gene transfer

International Journal of Pharmaceutics ◽

10.1016/j.ijpharm.2014.08.024 ◽

2014 ◽

Vol 475 (1-2) ◽

pp. 49-59 ◽

Cited By ~ 11

Author(s):

Soo-Yeon Kim ◽

Sang-Jin Lee ◽

Soo-Jeong Lim

Keyword(s):

Gene Transfer ◽

In Vivo Evaluation ◽

Adenoviral Gene Transfer

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Administration of cardiac mesenchymal cells modulates innate immunity in the acute phase of myocardial infarction in mice

Scientific Reports ◽

10.1038/s41598-020-71580-z ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Yi Kang ◽

Marjan Nasr ◽

Yiru Guo ◽

Shizuka Uchida ◽

Tyler Weirick ◽

...

Keyword(s):

Myocardial Infarction ◽

Innate Immunity ◽

Mechanism Of Action ◽

Cell Activation ◽

Cell Function ◽

Immune Cell ◽

Mesenchymal Cell ◽

Cell Types

Abstract Although cardiac mesenchymal cell (CMC) therapy mitigates post-infarct cardiac dysfunction, the underlying mechanisms remain unidentified. It is acknowledged that donor cells are neither appreciably retained nor meaningfully contribute to tissue regeneration—suggesting a paracrine-mediated mechanism of action. As the immune system is inextricably linked to wound healing/remodeling in the ischemically injured heart, the reparative actions of CMCs may be attributed to their immunoregulatory properties. The current study evaluated the consequences of CMC administration on post myocardial infarction (MI) immune responses in vivo and paracrine-mediated immune cell function in vitro. CMC administration preferentially elicited the recruitment of cell types associated with innate immunity (e.g., monocytes/macrophages and neutrophils). CMC paracrine signaling assays revealed enhancement in innate immune cell chemoattraction, survival, and phagocytosis, and diminished pro-inflammatory immune cell activation; data that identifies and catalogues fundamental immunomodulatory properties of CMCs, which have broad implications regarding the mechanism of action of CMCs in cardiac repair.

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An adenoviral gene transfer system to express high levels of soluble allogeneic MHC class I molecules in vitro and in vivo

Transplantation Proceedings ◽

10.1016/s0041-1345(02)03252-9 ◽

2002 ◽

Vol 34 (6) ◽

pp. 2318-2319

Author(s):

C Graeb ◽

M Justl ◽

S Tange ◽

K.-W Jauch ◽

E.K Geissler

Keyword(s):

Gene Transfer ◽

Mhc Class I ◽

Class I ◽

Transfer System ◽

Adenoviral Gene Transfer ◽

Gene Transfer System ◽

Mhc Class I Molecules

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Adenovirus capsid-display of the retro-oriented human complement inhibitor DAF reduces Ad vector–triggered immune responses in vitro and in vivo

Blood ◽

10.1182/blood-2010-03-276949 ◽

2010 ◽

Vol 116 (10) ◽

pp. 1669-1677 ◽

Cited By ~ 25

Author(s):

Sergey S. Seregin ◽

Yasser A. Aldhamen ◽

Daniel M. Appledorn ◽

Zachary C. Hartman ◽

Nathaniel J. Schuldt ◽

...

Keyword(s):

Gene Transfer ◽

Immune Responses ◽

Cell Activation ◽

Innate Immune ◽

Complement Inhibitor ◽

Human Complement ◽

Endothelial Cell Activation ◽

Capsid Display

Abstract Adenovirus (Ad) vectors are widely used in human clinical trials. However, at higher dosages, Ad vector–triggered innate toxicities remain a major obstacle to many applications. Ad interactions with the complement system significantly contribute to innate immune responses in several models of Ad-mediated gene transfer. We constructed a novel class of Ad vectors, genetically engineered to “capsid-display” native and retro-oriented versions of the human complement inhibitor decay-accelerating factor (DAF), as a fusion protein from the C-terminus of the Ad capsid protein IX. In contrast to conventional Ad vectors, DAF-displaying Ads dramatically minimized complement activation in vitro and complement-dependent immune responses in vivo. DAF-displaying Ads did not trigger thrombocytopenia, minimized endothelial cell activation, and had diminished inductions of proinflammatory cytokine and chemokine responses. The retro-oriented display of DAF facilitated the greatest improvements in vivo, with diminished activation of innate immune cells, such as dendritic and natural killer cells. In conclusion, Ad vectors can capsid-display proteins in a manner that not only retains the functionality of the displayed proteins but also potentially can be harnessed to improve the efficacy of this important gene transfer platform for numerous gene transfer applications.

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CircSMAD4 alleviates high glucose-induced inflammation, extracellular matrix deposition and apoptosis in mouse glomerulus mesangial cells by relieving miR-377-3p-mediated BMP7 inhibition

Diabetology & Metabolic Syndrome ◽

10.1186/s13098-021-00753-1 ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Rina Wu ◽

Zheli Niu ◽

Guangwei Ren ◽

Lin Ruan ◽

Lijun Sun

Keyword(s):

Extracellular Matrix ◽

High Glucose ◽

Mesangial Cells ◽

Circular Rna ◽

Inflammatory Factors ◽

Luciferase Reporter ◽

Protein Levels ◽

Matrix Deposition

Abstract Background Diabetic nephropathy (DN) is a common complication of diabetes mellitus. Accumulating studies suggest that the deregulation of circular RNA (circRNA) is involved in DN pathogenesis. This study aimed to investigate the role of circSMAD4 in DN models. Methods Mice were treated with streptozotocin to establish DN models in vivo. Mouse glomerulus mesangial cells (SV40-MES13) were treated with high glucose to establish DN models in vitro. The expression of circSMAD4, miR-377-3p and bone morphogenetic protein 7 (BMP7) mRNA was measured by quantitative real-time PCR (qPCR). The releases of inflammatory factors were examined by ELISA. The protein levels of fibrosis-related markers, apoptosis-related markers and BMP7 were checked by western blot. Cell apoptosis was monitored by flow cytometry assay. The predicted relationship between miR-377-3p and circSMAD4 or BMP7 was validated by dual-luciferase reporter assay or pull-down assay. Results CircSMAD4 was poorly expressed in DN mice and HG-treated SV40-MES13 cells. HG induced SV40-MES13 cell inflammation, extracellular matrix (ECM) deposition and apoptosis. CircSMAD4 overexpression alleviated, while circSMAD4 knockdown aggravated HG-induced SV40-MES13 cell injuries. MiR-377-3p was targeted by circSMAD4, and miR-377-3p enrichment partly reversed the effects of circSMAD4 overexpression. BMP7 was a target of miR-377-3p, and circSMAD4 regulated BMP7 expression by targeting miR-377-3p. MiR-377-3p overexpression aggravated HG-induced injuries by suppressing BMP7. Conclusion CircSMAD4 alleviates HG-induced SV40-MES13 cell inflammation, ECM deposition and apoptosis by relieving miR-377-3p-mediated inhibition on BMP7 in DN progression.

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Ultrasound targeted gene therapy in a mouse model of prostate cancer: Evaluation of immune response.

Journal of Clinical Oncology ◽

10.1200/jco.2018.36.6_suppl.120 ◽

2018 ◽

Vol 36 (6_suppl) ◽

pp. 120-120

Author(s):

Flavia De Carlo ◽

Litty Thomas ◽

Rounak Nande ◽

Olivia Boskovic ◽

Gailen Marshall ◽

...

Keyword(s):

Prostate Cancer ◽

Gene Transfer ◽

Cancer Cells ◽

Transduction Efficiency ◽

Acquired Immunity ◽

Prostate Cancer Cells ◽

Adenoviral Gene Transfer ◽

Specific Tissue

120 Background: Gene transfer to malignant sites using human adenoviruses (hAd) has been limited because of their immunogenicity. Murine cells often lack some of the receptors needed for hAd infection; therefore, are generally non-permissive for hAd infection and replication, which limits translational studies of adenoviral gene transfer techniques. We developed a gene transfer method, which uses a combination of lipid-encapsulated perfluorocarbon microbubbles (MBs) and ultrasound (US) to shield and deliver hAds to a specific tissue bypassing the requirement of the coxsackie and adenovirus receptor (CAR). Methods: Transduction efficiency and GFP protein expression of hAd.GFP was assessed by flow cytometry and fluorescence microscopy in murine TRAMP-C2 and human DU145 prostate cancer cells. Innate and acquired immunity response was determined by ELISA and CTL assay in C57BL/6 mice bearing TRAMP-C2 syngeneic tumor grafts following injections of MBs-Ad.GFP complexes in the presence or absence of ultrasound. Results: We observed that the murine prostate cancer cells TRAMP-C2 were transduced less efficiently by hAd.GFP than the human DU145 cells. We showed in vitro that the transduction rate was increased significantly in both TRAMP-C2 and DU145 prostate cancer cells when delivering the Ad particles by a combination of MBs and US. Moreover, we observed expression of the GFP transgene in both cell lines at 48 hours and 72 hours. Lack of activation of the innate and acquired immunity was observed in vivo by quantifying IL-6 and TNF-α cytokines, and by assaying neutralizing IgG antibodies and CTLs activity, following intratumoral or intravenous injections of MBs-Ad.GFP complexes in the presence or absence of ultrasound. Conclusions: This study demonstrates the feasibility of using the TRAMP-C2 murine model of prostate adenocarcinoma to translate our ultrasound-mediated MB-Ad delivery system from the bench to the clinic. Our data provides evidence that the TRAMP-C2 prostate cancer graft model is a suitable system to study in immune competent animals the capacity of lipid-encapsulated perfluorocarbon MBs and US, to shield and deliver hAds to a site-specific tissue bypassing the requirement of specific receptors.

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pVHL Function Is Essential for Endothelial Extracellular Matrix Deposition

Molecular and Cellular Biology ◽

10.1128/mcb.26.7.2519-2530.2006 ◽

2006 ◽

Vol 26 (7) ◽

pp. 2519-2530 ◽

Cited By ~ 63

Author(s):

Nan Tang ◽

Fiona Mack ◽

Volker H. Haase ◽

M. Celeste Simon ◽

Randall S. Johnson

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Critical Role ◽

Matrix Assembly ◽

Developmental Defects ◽

Vhl Gene ◽

Matrix Deposition ◽

Fibronectin Deposition

ABSTRACT The tumor suppressor von Hippel-Lindau protein (pVHL) is critical for cellular molecular oxygen sensing, acting to target degradation of the hypoxia-inducible factor alpha transcription factor subunits under normoxic conditions. We have found that independent of its function in regulating hypoxic response, the VHL gene plays a critical role in embryonic endothelium development through regulation of vascular extracellular matrix assembly. We created mice lacking the VHL gene in endothelial cells; these conditional null mice died at the same stage as homozygous VHL-null mice, with similar vascular developmental defects. These included defective vasculogenesis in the placental labyrinth, a collapsed endocardium, and impaired vessel network patterning. The defects in embryonic vascularization were correlated with a diminished vascular fibronectin deposition in vivo and defective endothelial extracellular fibronectin assembly in vitro. We found that the impaired migration and adhesion of VHL-null endothelial cells can be partially rescued by the addition of back exogenous fibronectin, which indicates that pVHL regulation of fibronectin deposition plays an important functional role in vascular patterning and maintenance of vascular integrity.

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GSK-3 Inhibition Modulates Metalloproteases in a Model of Lung Inflammation and Fibrosis

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.633054 ◽

2021 ◽

Vol 8 ◽

Author(s):

Francesco Cinetto ◽

Jessica Ceccato ◽

Ilaria Caputo ◽

Daniela Cangiano ◽

Barbara Montini ◽

...

Keyword(s):

Extracellular Matrix ◽

Lung Inflammation ◽

Therapeutic Strategy ◽

Glycogen Synthase ◽

Alveolar Cells ◽

Human Lung Fibroblast ◽

Matrix Deposition ◽

Extracellular Matrix Deposition

Idiopathic pulmonary fibrosis (IPF) is mainly characterized by aberrant extracellular matrix deposition, consequent to epithelial lung injury and myofibroblast activation, and inflammatory response. Glycogen synthase kinase 3 (GSK-3) is a serine–threonine kinase involved in several pathways, and its inhibition has been already suggested as a therapeutic strategy for IPF patients. There is evidence that GSK-3 is able to induce matrix metalloproteinase (MMP) expression and that its inhibition modulates MMP expression in the tissues. The aim of our study was to investigate the role of GSK-3 and its inhibition in the modulation of MMP-9 and -2 in an in vivo mouse model of lung fibrosis and in vitro using different cell lines exposed to pro-inflammatory or pro-fibrotic stimuli. We found that GSK-3 inhibition down-modulates gene expression and protein levels of MMP-9, MMP-2, and their inhibitors TIMP-1 and TIMP-2 in inflammatory cells harvested from bronchoalveolar lavage fluid (BALF) of mice treated with bleomycin as well as in interstitial alveolar macrophages and cuboidalized epithelial alveolar cells. To the same extent, GSK-3 inhibition blunted the increased MMP-9 and MMP-2 activity induced by pro-fibrotic stimuli in a human lung fibroblast cell line. Moreover, the αSMA protein level, a marker of fibroblast-to-myofibroblast transition involved in fibrosis, was decreased in primary fibroblasts treated with TGFβ following GSK-3 inhibition. Our results confirm the implication of GSK-3 in lung inflammation and fibrosis, suggesting that it might play its role by modulating MMP expression and activity but also pushing fibroblasts toward a myofibroblast phenotype and therefore enhancing extracellular matrix deposition. Thus, its inhibition could represent a possible therapeutic strategy.

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Extracellular Matrix and Fibrocyte Accumulation in BALB/c Mouse Lung upon Transient Overexpression of Oncostatin M

Cells ◽

10.3390/cells8020126 ◽

2019 ◽

Vol 8 (2) ◽

pp. 126 ◽

Cited By ~ 3

Author(s):

Fernando M. Botelho ◽

Rebecca Rodrigues ◽

Jessica Guerette ◽

Steven Wong ◽

Dominik K. Fritz ◽

...

Keyword(s):

Extracellular Matrix ◽

Cell Activation ◽

Adenovirus Vector ◽

Oncostatin M ◽

Mouse Lung ◽

Independent Manner ◽

Ecm Remodeling ◽

Transient Overexpression

The accumulation of extracellular matrix in lung diseases involves numerous factors, including cytokines and chemokines that participate in cell activation in lung tissues and the circulation of fibrocytes that contribute to local fibrotic responses. The transient overexpression of the gp130 cytokine Oncostatin M can induce extracellular matrix (ECM) accumulation in mouse lungs, and here, we assess a role for IL-13 in this activity using gene deficient mice. The endotracheal administration of an adenovirus vector encoding Oncostatin M (AdOSM) caused increases in parenchymal lung collagen accumulation, neutrophil numbers, and CXCL1/KC chemokine elevation in bronchioalveolar lavage fluids. These effects were similar in IL-13-/- mice at day 7; however, the ECM matrix induced by Oncostatin M (OSM) was reduced at day 14 in the IL-13-/- mice. CD45+col1+ fibrocyte numbers were elevated at day 7 due to AdOSM whereas macrophages were not. Day 14 levels of CD45+col1+ fibrocytes were maintained in the wildtype mice treated with AdOSM but were reduced in IL-13-/- mice. The expression of the fibrocyte chemotactic factor CXCL12/SDF-1 was suppressed marginally by AdOSM in vivo and significantly in vitro in mouse lung fibroblast cell cultures. Thus, Oncostatin M can stimulate inflammation in an IL-13-independent manner in BALB/c lungs; however, the ECM remodeling and fibrocyte accumulation is reduced in IL-13 deficiency.

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