The Experts below are selected from a list of 42813 Experts worldwide ranked by ideXlab platform
Dean J. Burkin - One of the best experts on this subject based on the ideXlab platform.
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Laminin-111 Improves the Anabolic Response to Mechanical Load in Aged Skeletal Muscle.
The journals of gerontology. Series A Biological sciences and medical sciences, 2020Co-Authors: Koyal Garg, Dean J. Burkin, Heather D. Huntsman, Ziad Mahmassani, Svyatoslav Dvoretskiy, M. Carmen Valero, Samuel Lapp, M S Stephen D Hauschka, Marni D. BoppartAbstract:Anabolic resistance to a mechanical stimulus may contribute to the loss of skeletal muscle mass observed with age. In this study, young and aged mice were injected with saline or human LM-111 (1 mg/kg). One week later, the myotendinous junction of the gastrocnemius muscle was removed via myotenectomy (MTE), thus placing a chronic mechanical stimulus on the remaining plantaris muscle for two weeks. LM-111 increased α7B integrin protein expression and clustering of the α7B integrin near DAPI + nuclei in aged muscle in response to MTE. LM-111 reduced CD11b + immune cells, enhanced repair, and improved the growth response to loading in aged plantaris muscle. These results suggest that LM-111 may represent a novel therapeutic approach to prevent and/or treat sarcopenia.
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Human Laminin-111 and Laminin-211 protein therapy prevents muscle disease progression in an immunodeficient mouse model of LAMA2-CMD.
Skeletal muscle, 2020Co-Authors: Pamela Barraza-flores, Hailey J. Hermann, Christina R. Bates, Tyler G. Allen, Timothy T. Grunert, Dean J. BurkinAbstract:Laminin-α2-related congenital muscular dystrophy (LAMA2-CMD) is a devastating genetic disease caused by mutations in the LAMA2 gene. These mutations result in progressive muscle wasting and inflammation leading to delayed milestones, and reduced lifespan in affected patients. There is currently no cure or treatment for LAMA2-CMD. Preclinical studies have demonstrated that mouse Laminin-111 can serve as an effective protein replacement therapy in a mouse model of LAMA2-CMD. In this study, we generated a novel immunocompromised dyW mouse model of LAMA2-CMD to study the role the immune system plays in muscle disease progression. We used this immune-deficient dyW mouse model to test the therapeutic benefits of recombinant human Laminin-111 and Laminin-211 protein therapy on Laminin-α2-deficient muscle disease progression. We show that immunodeficient Laminin-α2 null mice demonstrate subtle differences in muscle regeneration compared to immunocompetent animals during early disease stages but overall exhibit a comparable muscle disease progression. We found human Laminin-111 and Laminin-211 could serve as effective protein replacement strategies with mice showing improvements in muscle pathology and function. We observed that human Laminin-111 and Laminin-211 exhibit differences on satellite and myoblast cell populations and differentially affect muscle repair. This study describes the generation of a novel immunodeficient mouse model that allows investigation of the role the immune system plays in LAMA2-CMD. This model can be used to assess the therapeutic potential of heterologous therapies that would elicit an immune response. Using this model, we show that recombinant human Laminin-111 can serve as effective protein replacement therapy for the treatment of LAMA2-CMD.
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Laminin-111 protein therapy after disease onset slows muscle disease in a mouse model of Laminin-α2 related congenital muscular dystrophy.
Human molecular genetics, 2020Co-Authors: Pamela Barraza-flores, Katherine E. Bukovec, Marisela Dagda, Brandon W Conner, Ariany Oliveira-santos, Robert W. Grange, Dean J. BurkinAbstract:Laminin-α2 related congenital muscular dystrophy (LAMA2-CMD) is a fatal muscle disease caused by mutations in the LAMA2 gene. Laminin-α2 is critical for the formation of Laminin-211 and -221 heterotrimers in the muscle basal lamina. LAMA2-CMD patients exhibit hypotonia from birth and progressive muscle loss that results in developmental delay, confinement to a wheelchair, respiratory insufficiency and premature death. There is currently no cure or effective treatment for LAMA2-CMD. Several studies have shown Laminin-111 can serve as an effective protein-replacement therapy for LAMA2-CMD. Studies have demonstrated early treatment with Laminin-111 protein results in an increase in life expectancy and improvements in muscle pathology and function. Since LAMA2-CMD patients are often diagnosed after advanced disease, it is unclear if Laminin-111 protein therapy at an advanced stage of the disease can have beneficial outcomes. In this study, we tested the efficacy of Laminin-111 protein therapy after disease onset in a mouse model of LAMA2-CMD. Our results showed Laminin-111 treatment after muscle disease onset increased life expectancy, promoted muscle growth and increased muscle stiffness. Together these studies indicate Laminin-111 protein therapy either early or late in the disease process could serve as an effective protein replacement therapy for LAMA2-CMD.
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Human Laminin-111 and Laminin-211 protein therapy prevents muscle disease progression in an immune deficient mouse model of LAMA2-CMD
2020Co-Authors: Pamela Barraza-flores, Hailey J. Hermann, Christina R. Bates, Tyler G. Allen, Timothy T. Grunert, Dean J. BurkinAbstract:Abstract Background Laminin-α2 related Congenital Muscular dystrophy (LAMA2-CMD) is a devastating genetic disease caused by mutations in the LAMA2 gene. These mutations result in progressive muscle wasting and inflammation leading to delayed milestones, and reduced lifespan in affected patients. There is currently no cure or treatment for LAMA2-CMD. Preclinical studies have demonstrated that mouse Laminin-111 can serve as an effective protein replacement therapy in a mouse model of LAMA2-CMD. Methods In this study, we generated a novel immunocompromised dy W mouse model of LAMA2-CMD to study the role the immune system plays in muscle disease progression. We used this immune deficient dy W mouse model to test the therapeutic benefits of recombinant human Laminin-111 and Laminin-211 protein therapy on Laminin-α2 deficient muscle disease progression. Results We show that immune deficient Laminin-α2 null mice demonstrate subtle differences in muscle regeneration compared to immune competent animals during early disease stages, but overall exhibit a comparable muscle disease progression. We found human Laminin-111 and Laminin-211 could serve as effective protein replacement strategies with mice showing improvements in muscle pathology and function. We observed that human Laminin-111 and Laminin-211 exhibit differences on satellite and myoblast cell populations and differentially affect muscle repair. Conclusions This study describes the generation of a novel immune deficient mouse model that allows investigation of the role the immune system plays in LAMA2-CMD. This model can be used to assess the therapeutic potential of heterologous therapies that would illicit an immune response. Using this model, we show that recombinant human Laminin-111 can serve as effective protein replacement therapy for the treatment of LAMA2-CMD.
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Laminin and Integrin in LAMA2-Related Congenital Muscular Dystrophy: From Disease to Therapeutics.
Frontiers in molecular neuroscience, 2020Co-Authors: Pamela Barraza-flores, Ariany Oliveira-santos, Christina R. Bates, Dean J. BurkinAbstract:Laminin-α2-related congenital muscular dystrophy (LAMA2-CMD) is a devastating neuromuscular disease caused by mutations in the LAMA2 gene. These mutations result in the complete absence or truncated expression of the Laminin-α2 chain. The α2-chain is a major component of the Laminin-211 and Laminin-221 isoforms, the predominant Laminin isoforms in healthy adult skeletal muscle. Mutations in this chain result in progressive skeletal muscle degeneration as early as neonatally. Laminin-211/221 is a ligand for muscle cell receptors integrin-α7β1 and α-dystroglycan. LAMA2 mutations are correlated with integrin-α7β1 disruption in skeletal muscle. In this review, we will summarize Laminin-211/221 interactions with integrin-α7β1 in LAMA2-CMD muscle. Additionally, we will summarize recent developments using upregulation of Laminin-111 in the sarcolemma of Laminin-α2-deficient muscle. We will discuss potential mechanisms of action by which Laminin-111 is able to prevent myopathy. These published studies demonstrate that Laminin-111 is a disease modifier of LAMA2-CMD through different methods of delivery. Together, these studies show the potential for Laminin-111 therapy as a novel paradigm for the treatment of LAMA2-CMD.
Anne-gaëlle Borycki - One of the best experts on this subject based on the ideXlab platform.
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Basal lamina remodeling at the skeletal muscle stem cell niche mediates stem cell self-renewal
Nature communications, 2018Co-Authors: Shantisree Sandeepani Rayagiri, Daniele Ranaldi, Alexander Raven, Nur Izzah Farhana Mohamad Azhar, Olivier Lefebvre, Peter S. Zammit, Anne-gaëlle BoryckiAbstract:A central question in stem cell biology is the relationship between stem cells and their niche. Although previous reports have uncovered how signaling molecules released by niche cells support stem cell function, the role of the extra-cellular matrix (ECM) within the niche is unclear. Here, we show that upon activation, skeletal muscle stem cells (satellite cells) induce local remodeling of the ECM and the deposition of Laminin-α1 and Laminin-α5 into the basal lamina of the satellite cell niche. Genetic ablation of Laminin-α1, disruption of integrin-α6 signaling or blocking matrix metalloproteinase activity impairs satellite cell expansion and self-renewal. Collectively, our findings establish that remodeling of the ECM is an integral process of stem cell activity to support propagation and self-renewal, and may explain the effect Laminin-α1-containing supports have on embryonic and adult stem cells, as well as the regenerative activity of exogenous Laminin-111 therapy.
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Hedgehog signalling acts upstream of Laminin alpha1 transcription in the zebrafish paraxial mesoderm.
Matrix biology : journal of the International Society for Matrix Biology, 2016Co-Authors: Joseph Pickering, Vincent T. Cunliffe, Freek Van Eeden, Anne-gaëlle BoryckiAbstract:Laminin-111 (α1β1γ1) is a member of the Laminin family of extra-cellular matrix proteins that comprises 16 members, components of basement membranes. Laminin-111, one of the first Laminin proteins synthesised during embryogenesis, is required for basement membrane deposition and has essential roles in tissue morphogenesis and patterning. Yet, the mechanisms controlling Laminin-111 expression are poorly understood. We generated a zebrafish transgenic reporter line that reproduces faithfully the expression pattern of lama1, the gene encoding Laminin α1, and we used this reporter line to investigate lama1 transcriptional regulation. Our findings established that lama1 expression is controlled by intronic enhancers, including an enhancer directing expression in the paraxial mesoderm, anterior spinal cord and hindbrain, located in intron 1. We show that Hedgehog signalling is necessary and sufficient for lama1 transcription in the paraxial mesoderm and identify putative Gli/Zic binding sites that may mediate this control. These findings uncover a conserved role for Hedgehog signalling in the control of basement membrane assembly via its transcriptional regulation of lama1, and provide a mechanism to coordinate muscle cell fate specification in the zebrafish embryo.
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The myotomal basement membrane: insight into Laminin-111 function and its control by Sonic hedgehog signaling.
Cell adhesion & migration, 2013Co-Authors: Anne-gaëlle BoryckiAbstract:The importance of Laminin-containing basement membranes (BM) for adult muscle function is well established, in particular due to the severe phenotype of congenital muscular dystrophies in patients with mutations disrupting the BM-muscle cell interaction. Developing muscles in the embryo are also dependent on an intact BM. However, the processes controlled by BM-muscle cell interactions in the embryo are only beginning to be elucidated. In this review, we focus on the myotomal BM to illustrate the critical role of Laminin-111 in BM assembly and function at the surface of embryonic muscle cells. The myotomal BM provides also an interesting paradigm to study the complex interplay between Laminins-containing BM and growth factor-mediated signaling and activity.
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Sonic hedgehog-dependent synthesis of Laminin α1 controls basement membrane assembly in the myotome
Development (Cambridge England), 2009Co-Authors: Claire Anderson, Sólveig Thorsteinsdóttir, Anne-gaëlle BoryckiAbstract:Basement membranes have essential structural and signalling roles in tissue morphogenesis during embryonic development, but the mechanisms that control their formation are still poorly understood. Laminins are key components of basement membranes and are thought to be essential for initiation of basement membrane assembly. Here, we report that muscle progenitor cells populating the myotome migrate aberrantly in the ventral somite in the absence of sonic hedgehog (Shh) signalling, and we show that this defect is due to the failure to form a myotomal basement membrane. We reveal that expression of Lama1, which encodes Laminin alpha1, a subunit of Laminin-111, is not activated in Shh(-/-) embryos. Recovery of Lama1 expression or addition of exogenous Laminin-111 to Shh(-/-);Gli3(-/-) embryos restores the myotomal basement membrane, demonstrating that Laminin-111 is necessary and sufficient to initiate assembly of the myotomal basement membrane. This study uncovers an essential role for Shh signalling in the control of Laminin-111 synthesis and in the initiation of basement membrane assembly in the myotome. Furthermore, our data indicate that Laminin-111 function cannot be compensated by Laminin-511.
Koyal Garg - One of the best experts on this subject based on the ideXlab platform.
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Laminin-111 Improves the Anabolic Response to Mechanical Load in Aged Skeletal Muscle.
The journals of gerontology. Series A Biological sciences and medical sciences, 2020Co-Authors: Koyal Garg, Dean J. Burkin, Heather D. Huntsman, Ziad Mahmassani, Svyatoslav Dvoretskiy, M. Carmen Valero, Samuel Lapp, M S Stephen D Hauschka, Marni D. BoppartAbstract:Anabolic resistance to a mechanical stimulus may contribute to the loss of skeletal muscle mass observed with age. In this study, young and aged mice were injected with saline or human LM-111 (1 mg/kg). One week later, the myotendinous junction of the gastrocnemius muscle was removed via myotenectomy (MTE), thus placing a chronic mechanical stimulus on the remaining plantaris muscle for two weeks. LM-111 increased α7B integrin protein expression and clustering of the α7B integrin near DAPI + nuclei in aged muscle in response to MTE. LM-111 reduced CD11b + immune cells, enhanced repair, and improved the growth response to loading in aged plantaris muscle. These results suggest that LM-111 may represent a novel therapeutic approach to prevent and/or treat sarcopenia.
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Biomimetic sponges improve muscle structure and function following volumetric muscle loss
2020Co-Authors: Gabriel Haas, Andrew J. Dunn, Josh Madsen, Peter Genovese, Andrew Lin, Hannah Chauvin, Allison Paoli, Koyal GargAbstract:AbstractSkeletal muscle is inept in regenerating after traumatic injuries such as volumetric muscle loss (VML) due to significant loss of basal lamina and the resident satellite cells. Currently, there are no approved therapies for the treatment of muscle tissue following trauma. In this study, biomimetic sponges composed of gelatin, collagen, Laminin-111, and FK-506 were used for the treatment of VML in a rodent model. We observed that biomimetic sponge treatment improved muscle structure and function while modulating inflammation and limiting the extent of fibrotic tissue deposition. Specifically, sponge treatment increased the total number of myofibers, type 2B fiber cross-sectional area, myosin: collagen ratio, myofibers with central nuclei, and peak isometric torque compared to untreated VML injured muscles. As an acellular scaffold, biomimetic sponges provide a promising “off-the-shelf” clinical therapy for VML.
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the effect of Laminin 111 hydrogels on muscle regeneration in a murine model of injury
Tissue Engineering Part A, 2019Co-Authors: Madison Marcinczyk, Andrew J. Dunn, Gabriel Haas, Josh Madsen, Robert Scheidt, Krishna Patel, Muhamed Talovic, Koyal GargAbstract:Volumetric muscle loss (VML) is characterized by a critical loss of muscle tissue that is accompanied by severe functional impairment and often long-term disability. Clinical therapies currently us...
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The Effect of Laminin-111 Hydrogels on Muscle Regeneration in a Murine Model of Injury
Tissue engineering. Part A, 2018Co-Authors: Madison Marcinczyk, Andrew J. Dunn, Gabriel Haas, Josh Madsen, Robert Scheidt, Krishna Patel, Muhamed Talovic, Koyal GargAbstract:Extremity injuries make up the most common survivable injuries in vehicular accidents and modern military conflicts. A majority of these injuries involve volumetric muscle loss (VML). The potential for donor site morbidity may limit the clinical use of autologous muscle grafts for VML injuries. Treatments that can improve the regeneration of functional muscle tissue are critically needed to improve limb salvage and reduce the rate of delayed amputations. The development of a Laminin-111-enriched fibrin hydrogel will offer a potentially transformative and "off-the-shelf" clinically relevant therapy for functional skeletal muscle regeneration.
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Laminin-111 functionalized polyethylene glycol hydrogels support myogenic activity in vitro.
Biomedical materials (Bristol England), 2018Co-Authors: Natalia Ziemkiewicz, Madison Marcinczyk, Gabriel Haas, Josh Madsen, Robert Scheidt, Muhamed Talovic, Lindsay Hill, Anjali Patel, Silviya P. Zustiak, Koyal GargAbstract:Skeletal muscle has a remarkable regenerative capability following mild physical or chemical insult. However, following a critical loss of muscle tissue, the regeneration process is impaired due to the inadequate myogenic activity of muscle resident stem cells (i.e., satellite cells). Laminin (LM) is a heterotrimeric structural protein in the satellite cell niche that is crucial for maintaining its function. In this study, we created hydrogels composed of poly (ethylene glycol) (PEG) and LM-111 to provide an elastic substrate for satellite cell proliferation at the site of injury. The PEG-LM111 conjugates were mixed with 5% and 10% (w/v) pure PEG-diacrylate (PEGDA) and photopolymerized to form 5% and 10% PEGLM gels. Pure 5% and 10% PEGDA gels were used as controls. The modulus of both hydrogels containing 10% (w/v) PEGDA was significantly higher than the hydrogels containing 5% (w/v) PEGDA. The 5% PEGLM hydrogels showed significantly higher swelling in aqueous medium suggesting a more porous structure. C2C12 myoblasts cultured on the softer 5% PEGLM hydrogels showed a flat and spread-out morphology when compared to the rounded, multicell clusters formed on the 5% PEGDA, 10% PEGDA, and 10% PEGLM hydrogels. The 5% PEGLM hydrogels also promoted a significant increase in both vascular endothelial growth factor and interleukin-6 (IL-6) production from the myoblasts. Additionally, the expression of MyoD was significantly higher while that of myogenin and α-actinin trended higher on the 5% PEGLM hydrogels compared to 5% PEGDA on day 5. Our data suggests that the introduction of LM-111 into compliant PEG hydrogels promoted myoblast adhesion, survival, pro-regenerative growth factor production, and myogenic activity.
Mina J Bissell - One of the best experts on this subject based on the ideXlab platform.
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Generating a Fractal Microstructure of Laminin-111 to Signal to Cells.
Journal of visualized experiments : JoVE, 2020Co-Authors: Camilla Hochman-mendez, Ariel J Kent, Jamie L Inman, Mina J Bissell, Tatiana Coelho-sampaio, Claire RobertsonAbstract:Laminin-111 (Ln1) is an essential part of the extracellular matrix in epithelia, muscle and neural systems. We have previously demonstrated that the microstructure of Ln1 alters the way that it signals to cells, possibly because Ln1 assembly into networks exposes different adhesive domains. In this protocol, we describe three methods to generate polymerized Ln1.
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The microstructure of Laminin-111 compensates for dystroglycan loss in mammary epithelial cells in downstream expression of milk proteins.
Biomaterials, 2019Co-Authors: Ariel J Kent, Nasima Mayer, Camilla Hochman-mendez, Jamie L Inman, Mina J Bissell, Claire RobertsonAbstract:Laminin-111 (Ln-1), an extracellular matrix (ECM) glycoprotein found in the basement membrane of mammary gland epithelia, is essential for lactation. In mammary epithelial cells (MECs), dystroglycan (Dg) is believed to be necessary for polymerization of Laminin-111 into networks., thus we asked whether correct polymerization could compensate for Dg loss. Artificially polymerized Laminin-111 and the Laminin-glycoprotein mix Matrigel, both formed branching, spread networks with fractal dimensions from 1.7 to 1.8, whereas Laminin-111 in neutral buffers formed small aggregates without fractal properties (a fractal dimension of 2). In Dg knockout cells, either polymerized Laminin-111 or Matrigel readily attached to the cell surface, whereas aggregated Laminin-111 did not. In contrast, polymerized and aggregated Laminin-111 bound similarly to Dg knock-ins. Both polymerized Laminin-111 and Matrigel promoted cell rounding, clustering, formation of tight junctions, and expression of milk proteins, whereas aggregated Ln-1 did not attach to cells or promote functional differentiation. These findings support that the microstructure of Ln-1 networks in the basement membrane regulates mammary epithelial cell function.
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The microstructure of Laminin-111 compensates for dystroglycan loss in functional differentiation of mammary epithelial cells
bioRxiv, 2019Co-Authors: Ariel J Kent, Nasima Mayer, Camilla Hochman-mendez, Jamie L Inman, Mina J Bissell, Claire RobertsonAbstract:Abstract: Laminin-111, an extracellular matrix (ECM) glycoprotein found in the basement membrane of mammary gland epithelia, is essential for lactation. In mammary epithelial cells, dystroglycan (Dg) is believed to be necessary for polymerization of Laminin-111 into networks, thus we asked whether correct polymerization could compensate for Dg loss. Artificially polymerized Laminin-111 and the Laminin-glycoprotein mix Matrigel, both formed branching, spread networks with fractal dimensions from 1.7-1.8, whereas Laminin-111 in media formed small aggregates without fractal properties (a fractal dimension of 2). In Dg knockout cells, either polymerized Laminin-111 or Matrigel readily attached to the cell surface, whereas aggregated Laminin-111 did not. In contrast, polymerized and aggregated Laminin-111 bound similarly to Dg knock-ins. Both polymerized Laminin-111 and Matrigel promoted cell rounding, clustering, formation of tight junctions, and expression of milk proteins, whereas aggregated Ln-1 did not attach to cells or promote functional differentiation. Highlights Laminin assembles into a fractal network when in presence of either the cell surface receptor dystroglycan or acidic glycoproteins or an acidic buffer. When this microstructure is recreated with an acidic treatment, Laminin binds readily to dystroglycan null cells and induces functional differentiation of mammary epithelial cells.
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Laminin-111 and the Level of Nuclear Actin Regulate Epithelial Quiescence via Exportin-6.
Cell reports, 2017Co-Authors: Ana Paula Zen Petisco Fiore, Mina J Bissell, Virginia A. Spencer, Hidetoshi Mori, Hernandes F. Carvalho, Alexandre Bruni-cardosoAbstract:Nuclear actin (N-actin) is known to participate in the regulation of gene expression. We showed previously that N-actin levels mediate the growth and quiescence of mouse epithelial cells in response to Laminin-111 (LN1), a component of the mammary basement membrane (BM). We know that BM is defective in malignant cells, and we show here that it is the LN1/N-actin pathway that is aberrant in human breast cancer cells, leading to continuous growth. Photobleaching assays revealed that N-actin exit in nonmalignant cells begins as early as 30 min after LN1 treatment. LN1 attenuates the PI3K pathway leading to upregulation of exportin-6 (XPO6) activity and shuttles actin out of the nucleus. Silencing XPO6 prevents quiescence. Malignant cells are impervious to LN1 signaling. These results shed light on the crucial role of LN1 in quiescence and differentiation and how defects in the LN1/PI3K/XPO6/N-actin axis explain the loss of tissue homeostasis and growth control that contributes to malignant progression.
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Laminin regulates PI3K basal localization and activation to sustain STAT5 activation
Cell cycle (Georgetown Tex.), 2010Co-Authors: Virginia A. Spencer, Dinah Levy Groesser, Mina J BissellAbstract:Extracellular matrix (ECM) is a key regulator of tissue morphogenesis and functional differentiation in the mammary gland. We showed recently that Laminin-111 (LN1) together with prolactin induces β-casein expression in mammary epithelial cells (MECs) by sustaining STAT5 activation. Others have shown that Rac1 is required for integrin-mediated STAT5 activation, but molecules upstream of Rac1 remain to be elucidated. Here, we show that exposure to three-dimensional (3D), Laminin-rich ECM (LrECM) gels changes the localization of phosphoinositide 3-kinase (PI3K) in MECs from diffuse to basal, accompanied by the activation of the PI3K-Rac1 signaling pathway. We show by co-immunoprecipitation that Rac1 associates with STAT5 and that LrECM treatment enhances this interaction. Blocking PI3K with LY294002 inhibits LrECM-dependent Rac1 activation, attenuates sustained STAT5 phosphorylation and blocks β-casein gene transcription. These results indicate that PI3K is a key mediator of the LN1-induced signaling cascade, which controls the activity of transcription factors essential for tissue-specific gene expression.
Motoyoshi Nomizu - One of the best experts on this subject based on the ideXlab platform.
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effects of Laminin 111 peptide coatings on rat neural stem progenitor cell culture
Experimental Cell Research, 2021Co-Authors: Hideki Hayashi, Yuji Yamada, Norio Takagi, Ichiro Horinokita, Keisuke Hamada, Motoyoshi NomizuAbstract:Neurons require adhesive scaffolds for their growth and differentiation. Laminins are a major cell adhesive component of basement membranes and have various biological activities in the peripheral and central nervous systems. Here, we evaluated the biological activities of 5 peptides derived from Laminin-111 as a scaffold for mouse neuroblastoma Neuro2a cells and rat neural stem/progenitor cells (NPCs). The 5 peptides showed Neuro2a cell attachment activity similar to that of poly-d-lysine. However, when NPCs were cultured on the peptides, 2 syndecan-binding peptides, AG73 (RKRLQVQLSIRT, mouse Laminin α1 chain 2719-2730) and C16 (KAFDITYVRLKF, Laminin γ1 chain 139-150), demonstrated significantly higher cell attachment and neurite extension activities than other peptides including integrin-binding ones. Long-term cell culture experiments showed that both AG73 and C16 supported the growth of neurons and astrocytes that had differentiated from NPCs. Furthermore, C16 markedly promoted the expression of neuronal markers such as synaptosomal-associated protein-25 and syntaxin 1A. These results indicate that AG73 and C16 are useful for NPC cultures and that C16 can be applied to specialized research on synapses in differentiated neurons. These peptides have the potential for use as valuable biomaterials for NPC research.
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Effects of Laminin-111 peptide coatings on rat neural stem/progenitor cell culture.
Experimental cell research, 2020Co-Authors: Hideki Hayashi, Yuji Yamada, Norio Takagi, Ichiro Horinokita, Keisuke Hamada, Motoyoshi NomizuAbstract:Neurons require adhesive scaffolds for their growth and differentiation. Laminins are a major cell adhesive component of basement membranes and have various biological activities in the peripheral and central nervous systems. Here, we evaluated the biological activities of 5 peptides derived from Laminin-111 as a scaffold for mouse neuroblastoma Neuro2a cells and rat neural stem/progenitor cells (NPCs). The 5 peptides showed Neuro2a cell attachment activity similar to that of poly-d-lysine. However, when NPCs were cultured on the peptides, 2 syndecan-binding peptides, AG73 (RKRLQVQLSIRT, mouse Laminin α1 chain 2719-2730) and C16 (KAFDITYVRLKF, Laminin γ1 chain 139-150), demonstrated significantly higher cell attachment and neurite extension activities than other peptides including integrin-binding ones. Long-term cell culture experiments showed that both AG73 and C16 supported the growth of neurons and astrocytes that had differentiated from NPCs. Furthermore, C16 markedly promoted the expression of neuronal markers such as synaptosomal-associated protein-25 and syntaxin 1A. These results indicate that AG73 and C16 are useful for NPC cultures and that C16 can be applied to specialized research on synapses in differentiated neurons. These peptides have the potential for use as valuable biomaterials for NPC research.
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Biological activities of Laminin-111-derived peptide-chitosan matrices in a primary culture of rat cortical neurons.
Archives of biochemistry and biophysics, 2018Co-Authors: Hideki Hayashi, Mariko Yamada, Jun Kumai, Norio Takagi, Motoyoshi NomizuAbstract:Cell adhesive biomaterials have been used for various cells in culture, especially for primary cultures of neurons. Here we examined Laminin-111 and its active peptides conjugated to chitosan matrices (ChtMs) for primary culture of rat cortical neurons. Laminin-111 on poly-d-lysine substrate promoted neuronal cell attachment and differentiation. The biological activity of six active Laminin-111-derived peptides was examined using a peptide-ChtM construct. When the syndecan-binding peptides, AG73 (RKRLQVQLSIRT, mouse Laminin α1 chain 2719-2730) and C16 (KAFDITYVRLKF, Laminin γ1 chain 139-150), were conjugated to chitosan, AG73-ChtM and C16-ChtM showed potent neuronal cell attachment activity and promoted axon extension by primary cultured rat cortical neurons. However, the remaining peptides, including integrin-binding peptides, did not show activity when conjugated to ChtM. AG73-ChtM and C16-ChtM also supported neuron survival for at least 4 weeks in serum-free medium without a glia feeder layer. These data suggest that AG73-ChtM and C16-ChtM are useful for primary cultures of central nervous system neurons and have a potential for use as functional biomaterials for tissue engineering in the central nervous system.
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Laminin-111-derived peptide-hyaluronate hydrogels as a synthetic basement membrane.
Biomaterials, 2013Co-Authors: Yuji Yamada, Yamato Kikkawa, Kentaro Hozumi, Fumihiko Katagiri, Motoyoshi NomizuAbstract:We have identified a number of cell-adhesive peptides from Laminins, a major component of basement membranes. Cell-adhesive peptides derived from basement membrane proteins are potential candidates for incorporating cell-binding activities into scaffold materials for tissue engineering. Our goal is development of a chemically synthetic basement membrane using Laminin-derived cell-adhesive peptides and polymeric materials. In this study, we used hyaluronic acid (HA) as a scaffold material and Laminin-derived cell-adhesive peptides, A99 (AGTFALRGDNPQG, binds to integrin αvβ3), AG73 (RKRLQVQLSIRT, binds to syndecans), and an A99/AG73 mixture (molar ratio = 9:1) conjugated to two-dimensional (2D) HA matrices. As a result, it was found that the 2D A99/AG73-HA matrices have strong biological functions, such as cell attachment, cell spreading, and neurite outgrowth, similar to that of basement membrane extract (BME)-coated plates. Next, we developed three-dimensional (3D) peptide-HA matrices using the A99/AG73 mixture. The 3D A99/AG73-HA matrices promoted cell spreading and improved cell viability and collagen gene expression. Further, PC12 neurite extension was observed in the 3D A99/AG73-HA matrices. These biological activities of the 3D A99/AG73-HA matrices were similar to those of the 3D BME matrices. These results suggest that the peptide-HA matrices are useful as 2D and 3D matrices and can be applied for tissue engineering as a synthetic basement membrane.
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Laminin-111-derived peptides and cancer
Cell adhesion & migration, 2012Co-Authors: Yamato Kikkawa, Kentaro Hozumi, Fumihiko Katagiri, Motoyoshi Nomizu, Hynda K. Kleinman, Jennifer E. KoblinskiAbstract:Laminin-111 is a large trimeric basement membrane glycoprotein with many active sites. In particular, four peptides active in tumor malignancy studies have been identified in Laminin-111 using a systematic peptide screening method followed by various assays. Two of the peptides (IKVAV and AG73) are found on the α1 chain, one (YIGSR) of the β1 chain and one (C16) on the γ1 chain. The four peptides have distinct activities and receptors. Since three of the peptides (IKVAV, AG73 and C16) strongly promote tumor growth, this may explain the potent effects Laminin-111 has on malignant cells. The peptide, YIGSR, decreases tumor growth and experimental metastasis via a 32/67 kD receptor while IKVAV increases tumor growth, angiogenesis and protease activity via integrin receptors. AG73 increases tumor growth and metastases via syndecan receptors. C16 increases tumor growth and angiogenesis via integrins. Identification of such sites on Laminin-111 will have use in defining strategies to develop therapeutics for ca...
