scholarly journals Functional Expression of Mouse Relaxin and Mouse Relaxin-3 in the Lung from an Ebola Virus Glycoprotein-Pseudotyped Lentivirus via Tracheal Delivery

Endocrinology ◽  
2006 ◽  
Vol 147 (8) ◽  
pp. 3797-3808 ◽  
Author(s):  
Josh D. Silvertown ◽  
Jagdeep S. Walia ◽  
Alastair J. Summerlee ◽  
Jeffrey A. Medin
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Ebola Virus ◽  
Peptide Hormone ◽  
Biologically Active ◽  
Extracellular Matrix Protein ◽  
Mouse Lung ◽  
Protein Accumulation ◽  
Virus Glycoprotein ◽  
Age Related

The peptide hormone relaxin is a known modulator of connective tissue and the extracellular matrix by virtue of its ability to regulate matrix metalloproteinases (MMPs). Relaxin knockout mice exhibit age-related pulmonary fibrosis, and delivery of recombinant human H2 relaxin ameliorates fibrotic-like conditions in the mouse lung. We investigated whether lentiviral vectors (LVs) engineering the expression of murine relaxins could induce MMP activity in the mouse lung. Mouse relaxin and mouse relaxin-3 peptides engineered by recombinant LVs were biologically active as shown by stimulation of cAMP from both THP-1 and 293T cells stably expressing relaxin receptor LGR7 and by up-regulation of MMP-2 activity from primary C57BL/6 lung cell cultures. To provide the virions with enhanced tropism for the lung, LVs were pseudotyped with the Zaire strain of the Ebola virus glycoprotein (EboZ GP) and delivered by endotracheal intubation. LVs engineering luciferase pseudotyped with EboZ GP, but not with vesicular stomatitis virus glycoprotein resulted in successful LV transduction and transgene expression in C57BL/6 mouse lung by as early as d 4. Mice treated via tracheal delivery with EboZ GP pseudotyped LVs that engineered expression of mouse relaxins exhibited increased MMP-2 and MMP-9 activity in lung tissue up until the end of our study at d 21. Taken together, this study provides proof-of- principle that relaxin gene expression targeted to the mouse lungs can result in enhanced MMP activity offering potential for alleviating disease conditions characterized by dysregulation of extracellular matrix protein accumulation.

A novel age-related venous amyloidosis derived from EGF-containing fibulin-like extracellular matrix protein 1

2018 ◽  
Vol 247 (4) ◽  
pp. 444-455 ◽  
Author(s):  
Masayoshi Tasaki ◽  
Mitsuharu Ueda ◽  
Yoshinobu Hoshii ◽  
Mayumi Mizukami ◽  
Sayaka Matsumoto ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Extracellular Matrix Protein ◽  
Extracellular Matrix Protein 1 ◽  
Age Related

Effect of Age and Frequency on Intervertebral Disc Cell Response to Dynamic Compression

2007 ◽  
Author(s):  
Casey L. Korecki ◽  
Catherine K. Kuo ◽  
Rocky S. Tuan ◽  
James C. Iatridis
Keyword(s):  
Extracellular Matrix ◽  
Intervertebral Disc ◽  
Mechanical Stimulation ◽  
Matrix Protein ◽  
Dynamic Compression ◽  
Model Systems ◽  
Extracellular Matrix Protein ◽  
Tissue Culture Cell ◽  
Protein Accumulation

The intervertebral disc (IVD) is a unique orthopaedic tissue consisting of at least two cell types: fibroblast-like annulus fibrosus (AF) cells and chondrocyte-like nucleus pulposus (NP) cells. Culture of cells in 3D gel matrices (such as alginate or agarose), maintains the normal morphology and ECM molecule production of chondrocytes for extended periods of time and also allows the application of various forms of mechanical stimulation, such as hydrostatic or compressive loading. In vivo studies have shown IVD cells to be responsive to frequency, duration, and amplitude of mechanical load [1]. IVD literature on mechanobiology uses varying methodologies to apply dynamic loads (compression, hydrostatic forces), with different times of mechanical stimulation, differences in model systems (in vivo, tissue culture, cell culture), species, and ages, and an optimal loading protocol to stimulate extracellular matrix protein accumulation is unknown. The overall goal of this work is to evaluate the potential, and perhaps even feasibility, of mechanical stimulation for extracellular matrix (ECM) regeneration using intervertebral disc cells. Also of interest is whether cells from mature tissue are capable of serving as a potential cell source for future IVD regeneration [2,3].

scholarly journals Evolutionarily conserved sequence motif analysis guides development of chemically defined hydrogels for therapeutic vascularization

2020 ◽  
Vol 6 (28) ◽  
pp. eaaz5894
Author(s):  
Jia Jia ◽  
Eun Je Jeon ◽  
Mei Li ◽  
Dylan J. Richards ◽  
Soojin Lee ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Network Formation ◽  
Blood Perfusion ◽  
Biologically Active ◽  
Extracellular Matrix Protein ◽  
Sequence Motif ◽  
Motif Analysis ◽  
Conserved Sequence ◽  
Evolutionarily Conserved

Biologically active ligands (e.g., RGDS from fibronectin) play critical roles in the development of chemically defined biomaterials. However, recent decades have shown only limited progress in discovering novel extracellular matrix–protein–derived ligands for translational applications. Through motif analysis of evolutionarily conserved RGD-containing regions in laminin (LM) and peptide-functionalized hydrogel microarray screening, we identified a peptide (a1) that showed superior supports for endothelial cell (EC) functions. Mechanistic studies attributed the results to the capacity of a1 engaging both LM- and Fn-binding integrins. RNA sequencing of ECs in a1-functionalized hydrogels showed ~60% similarities with Matrigel in “vasculature development” gene ontology terms. Vasculogenesis assays revealed the capacity of a1-formulated hydrogels to improve EC network formation. Injectable alginates functionalized with a1 and MMPQK (a vascular endothelial growth factor–mimetic peptide with a matrix metalloproteinase–degradable linker) increased blood perfusion and functional recovery over decellularized extracellular matrix and (RGDS + MMPQK)–functionalized hydrogels in an ischemic hindlimb model, illustrating the power of this approach.

1280: Increased Susceptibility to Genitovaginal Prolapse Associated with a Polymorphism in the Promoter of the Extracellular Matrix Protein LAMC1

2007 ◽  
Vol 177 (4S) ◽  
pp. 421-422
Author(s):  
Ganka Nikolova ◽  
Christian O. Twiss ◽  
Hane Lee ◽  
Nelson Stanley ◽  
Janet Sinsheimer ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Extracellular Matrix Protein ◽  
Increased Susceptibility

scholarly journals Homogenous overexpression of the extracellular matrix protein Netrin-1 in a hollow fiber bioreactor

2021 ◽  
Author(s):  
Aniel Moya-Torres ◽  
Monika Gupta ◽  
Fabian Heide ◽  
Natalie Krahn ◽  
Scott Legare ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hollow Fiber ◽  
Mammalian Cells ◽  
Matrix Protein ◽  
Continuous Production ◽  
Extracellular Matrix Protein ◽  
Close Monitoring ◽  
High Quality ◽  
Biolayer Interferometry ◽  
Hollow Fiber Bioreactor

Abstract The production of recombinant proteins for functional and biophysical studies, especially in the field of structural determination, still represents a challenge as high quality and quantities are needed to adequately perform experiments. This is in part solved by optimizing protein constructs and expression conditions to maximize the yields in regular flask expression systems. Still, work flow and effort can be substantial with no guarantee to obtain improvements. This study presents a combination of workflows that can be used to dramatically increase protein production and improve processing results, specifically for the extracellular matrix protein Netrin-1. This proteoglycan is an axon guidance cue which interacts with various receptors to initiate downstream signaling cascades affecting cell differentiation, proliferation, metabolism, and survival. We were able to produce large glycoprotein quantities in mammalian cells, which were engineered for protein overexpression and secretion into the media using the controlled environment provided by a hollow fiber bioreactor. Close monitoring of the internal bioreactor conditions allowed for stable production over an extended period of time. In addition to this, Netrin-1 concentrations were monitored in expression media through biolayer interferometry which allowed us to increase Netrin-1 media concentrations tenfold over our current flask systems while preserving excellent protein quality and in solution behavior. Our particular combination of genetic engineering, cell culture system, protein purification, and biophysical characterization permitted us to establish an efficient and continuous production of high-quality protein suitable for structural biology studies that can be translated to various biological systems. Key points • Hollow fiber bioreactor produces substantial yields of homogenous Netrin-1 • Biolayer interferometry allows target protein quantitation in expression media • High production yields in the bioreactor do not impair Netrin-1 proteoglycan quality Graphical abstract

Direct interaction of the extracellular matrix protein DANCE with apolipoprotein(a) mediated by the kringle IV-type 2 domain

2002 ◽  
Vol 267 (4) ◽  
pp. 440-446 ◽  
Author(s):  
A. Kapetanopoulos ◽  
F. Fresser ◽  
G. Millonig ◽  
Y. Shaul ◽  
G. Baier ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Direct Interaction ◽  
Extracellular Matrix Protein ◽  
Apolipoprotein A
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