YIGSR Modified - Laminin Functional Hydrogel
Laminins are extracellular matrix components of the basement membrane that play a critical role in tissue development and function. Laminins are ubiquitously expressed and have multiple biological and pathological functions, including cell adhesion, cell differentiation, cell migration, neurite outgrowth, and cancer cell metastasis. Laminins consist of α, β, and γ subunits, which assemble to form a cruciform heteromeric glycoprotein1, 2. The laminin β1 chain has seven structurally distinct domains and several important active sites that have been identified and used to produce synthetic peptides with functional readouts3. One such synthetic peptide, YIGSR (Tyr–Ile–Gly–Ser–Arg), which consists of residues 929–933 on the β1 chain, has multiple functions. This pentapeptide can bind to laminin receptors to activate signaling pathways and promote cell adhesion4.
YIGSR synthetic peptides are commonly used to promote cell attachment and may be used cooperatively with a variety of different biomimetic materials, including three-dimensional tunable hydrogels (VitroGel YIGRS, VitroGel LDP1, VitroGel LDP2). These peptides can facilitate cell adhesion, similar to laminin, in a number of cell types, such as cancer cells, fibroblasts, neurons, and glia, without altering the physiological response5. Promotion of cell spreading and colocalization with laminin receptors and adhesion signaling proteins has been observed in the presence of immobilized, covalently attached YIGSR4. When coated on a number of different surfaces, such as polystyrene, YIGSR can influence cell attachment and spreading of different cancer cell lines, such as mouse melanoma and human rhabdomyosarcoma cells6. Multiple other effects of YIGSR on cell lines have been shown as well; for example, YIGSR increased collagen synthesis in Hs27 human dermal fibroblasts7. Furthermore, endotheliocytes were shown to migrate toward a gradient of YIGSR, demonstrating the value of YIGSR in cell migration studies8.
More recently, synthetic peptides, such as YIGSR, have been used to aid in creating three-dimensional stem cell spheroids and hydrogels in order to mimic a variety of tissues. By recapitulate a basement membrane-like environment, researchers have been able to induce asymmetric stem cell division in order to form stratified tissue-like spheroids9,10. In a recent study, YIGSR was tethered to hyperbranched poly-lysine peptides to form an environment conducive to stem cell survival and asymmetric division10. This environment allowed for the formation of three-dimensional human bone marrow mesenchymal stem cell spheroids. This basement membrane-like microenvironment leads to the clustering of integrin receptors, activation of intracellular signaling pathways, and regulation of the cytoskeleton. In addition to the spheroid formation, YIGSR may be used to study a number of cellular processes, such as cell survival, differentiation, morphogenesis, and regeneration, via covalent immobilization in three-dimensional hydrogels. Recently, a study using polyethylene glycol norbornene as a biomaterial made use of YIGSR as an adhesive substratum11. The researchers used YIGSR to investigate the combinatory effects of a three-dimensional environment and soluble biochemical cues on myofibrogenesis of human mesenchymal stromal cells. Moreover, other studies have tested the ability of YIGSR to act as both an adhesive source and cell survival factor for stem cells and neural progenitors in three-dimensional environments12,13. In another study utilizing three-dimensional matrices, YIGSR promoted the formation of compact aggregates of human pancreatic ductal epithelial cells14. These data illustrate the utility of YIGSR in studying a wide variety of biological processes in spheroids and three-dimensional hydrogel systems.
The extracellular matrix plays a critical role in neuronal differentiation and neuron morphogenesis. Immobilized, covalently bound YIGSR has proven to be a useful tool in studying these processes in two- and three-dimensional environments. YIGSR enhances neuronal cell attachment15. Nanofibers, functionalized with YIGSR, were shown to significantly increase differentiation of mouse embryonic stem cells into mature neurons, as well as promote neurite extension16. Furthermore, when used in conjunction with RGD peptide, YIGSR regulates neuronal differentiation and morphogenesis of PC12 cells in vitro17. Additional studies have used YIGSR to functionalize a number of different nanofiber and hydrogel systems further illustrating its ability to promote neuronal differentiation and regulate morphogenesis and neurite formation18-20.
The immobilization, via covalent attachment to a biomaterial or surface, of YIGSR is required to yield the effects observed on cell attachment, spreading, and migration. In contrast, soluble, or non-covalently bound, YIGSR acts as an inhibitor to these cellular processes, due to laminin receptor competition. For example, human neuroblastoma cell attachment to laminin was inhibited by YIGSR, due to competitive binding21. Additionally, YIGSR was found to inhibit laminin induced hypercontractility of airway smooth muscle cells in both in vitro and in vivo models of asthma22. The ability of soluble YIGSR to act in an inhibitory manner, due to laminin receptor competition, has been used in a number of studies attempting to influence tumor growth, cancer cell invasion, and metastasis. Basement membrane extracts, such as Matrigel, and glycoproteins, such as laminin, create an attractive environment for cancer cell invasion. Furthermore, the 32/67 kDa laminin receptor correlates positively with tumor malignancy23. However, non-covalently bound YIGSR can bind to this laminin receptor resulting in reduced metastasis and tumor enlargement. In mice injected with melanoma cells, YIGSR was found to strongly reduce lung colony formation24. YIGSR inhibits growth and metastasis of melanoma and leukemic cells25. Human pre-B acute lymphoblastic leukemia cell colony formation and invasion was inhibited in the presence of YIGSR26. Laminin induced cell attachment of RCT sarcoma cells was decreased with YIGSR27. Additionally, YIGSR has been found to promote fibrosarcoma cell apoptosis28. Furthermore, when conjugated with polyvinyl pyrrolidone, YIGSR, showed exceptional anti-metastatic effects in B16-BL6 melanoma cells29. These data illustrate the utility of soluble, non-covalently bound YIGSR as a metastatic and tumor growth inhibitor.
Multiple studies have further demonstrated the usefulness of YIGSR by investigating, and comparing, the effects of this peptide with the RGD peptide. Non-covalent binding of YIGSR and RGD was shown to have synergistic positive effects on sciatic nerve generation30. Moreover, inhibitory effects on cell adhesion have been observed upon application of YIGSR and RGD in the presence of the basement membrane molecule laminin and fibronectin, respectively, due to receptor competition31. The synergistic effects of YIGSR and RGD have also been demonstrated in hydrogels. A recent study aimed at optimizing neuronal differentiation in three-dimensional hydrogels found an increase in the number of mature neurons, differentiated from induced pluripotent stem cells, in hydrogels treated with these peptides32. Multiple other studies have compared biological responses to YIGSR and RGD and found differential effects on cell adhesion5, cell mobility and migration33, 34, and tissue engineering35, 36. Taken together these data display the combinatory, and contrasting, effects of YIGSR and RGD on a number of processes and different cell types.
Cell Type Behavior Reference Table for VitroGel YIGSR
|Human vein endotheliocytes||Increased cell migration|
|Human endothelial||Promoted cell differentiation|
|Human umbilical vein endothelial cells||Upregulation in gene expression|
|Rat neonatal cardiac||Promoted cell attachment similar to laminin|
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- Venstrom, K.A. and L.F. Reichardt, Extracellular matrix. 2: Role of extracellular matrix molecules and their receptors in the nervous system. FASEB J, 1993. 7(11): p. 996-1003.
- Aumailley, M., The laminin family. Cell Adh Migr, 2013. 7(1): p. 48-55.
- Massia, S.P., S.S. Rao, and J.A. Hubbell, Covalently immobilized laminin peptide Tyr-Ile-Gly-Ser-Arg (YIGSR) supports cell spreading and co-localization of the 67-kilodalton laminin receptor with alpha-actinin and vinculin. J Biol Chem, 1993. 268(11): p. 8053-9.
- Boateng, S.Y., et al., RGD and YIGSR synthetic peptides facilitate cellular adhesion identical to that of laminin and fibronectin but alter the physiology of neonatal cardiac myocytes. Am J Physiol Cell Physiol, 2005. 288(1): p. C30-8.
- Maeda, T., K. Titani, and K. Sekiguchi, Cell-adhesive activity and receptor-binding specificity of the laminin-derived YIGSR sequence grafted onto Staphylococcal protein A. J Biochem, 1994. 115(2): p. 182-9.
- Yoon, J.H., et al., Laminin peptide YIGSR induces collagen synthesis in Hs27 human dermal fibroblasts. Biochem Biophys Res Commun, 2012. 428(3): p. 416-21.
- Ren, T., et al., Complementary density gradient of Poly(hydroxyethyl methacrylate) and YIGSR selectively guides migration of endotheliocytes. Biomacromolecules, 2014. 15(6): p. 2256-64.
- Spradling, A., D. Drummond-Barbosa, and T. Kai, Stem cells find their niche. Nature, 2001. 414(6859): p. 98-104.
- Perugini, V., et al., Hyperbranched poly(-lysine) substrate presenting the laminin sequence YIGSR induces the formation of spheroids in adult bone marrow stem cells. PLoS One, 2017. 12(12): p. e0187182.
- Usprech, J., et al., Combinatorial screening of 3D biomaterial properties that promote myofibrogenesis for mesenchymal stromal cell-based heart valve tissue engineering. Acta Biomater, 2017. 58: p. 34-43.
- Ovadia, E.M., D.W. Colby, and A.M. Kloxin, Designing well-defined photopolymerized synthetic matrices for three-dimensional culture and differentiation of induced pluripotent stem cells. Biomater Sci, 2018. 6(6): p. 1358-1370.
- Zhao, T., et al., Tunable, Injectable Hydrogels Based on Peptide-Cross-Linked, Cyclized Polymer Nanoparticles for Neural Progenitor Cell Delivery. Biomacromolecules, 2017. 18(9): p. 2723-2731.
- Raza, A., C.S. Ki, and C.C. Lin, The influence of matrix properties on growth and morphogenesis of human pancreatic ductal epithelial cells in 3D. Biomaterials, 2013. 34(21): p. 5117-27.
- Ranieri, J.P., et al., Neuronal cell attachment to fluorinated ethylene propylene films with covalently immobilized laminin oligopeptides YIGSR and IKVAV. II. J Biomed Mater Res, 1995. 29(6): p. 779-85.
- Callahan, L.A., et al., Directed differentiation and neurite extension of mouse embryonic stem cell on aligned poly(lactide) nanofibers functionalized with YIGSR peptide. Biomaterials, 2013. 34(36): p. 9089-95.
- Lee, J.W. and K.Y. Lee, Dual peptide-presenting hydrogels for controlling the phenotype of PC12 cells. Colloids Surf B Biointerfaces, 2017. 152: p. 36-41.
- Manchineella, S., et al., Surface-Functionalized Silk Fibroin Films as a Platform To Guide Neuron-like Differentiation of Human Mesenchymal Stem Cells. ACS Appl Mater Interfaces, 2016. 8(35): p. 22849-59.
- Silantyeva, E.A., et al., Accelerated neural differentiation of mouse embryonic stem cells on aligned GYIGSR-functionalized nanofibers. Acta Biomater, 2018. 75: p. 129-139.
- Cui, G.H., et al., Designer Self-Assemble Peptides Maximize the Therapeutic Benefits of Neural Stem Cell Transplantation for Alzheimer’s Disease via Enhancing Neuron Differentiation and Paracrine Action. Mol Neurobiol, 2016. 53(2): p. 1108-1123.
- Bushkin-Harav, I., N.B. Garty, and U.Z. Littauer, Down-regulation of a 67-kDa YIGSR-binding protein upon differentiation of human neuroblastoma cells. J Biol Chem, 1995. 270(22): p. 13422-8.
- Dekkers, B.G., et al., The laminin beta1-competing peptide YIGSR induces a hypercontractile, hypoproliferative airway smooth muscle phenotype in an animal model of allergic asthma. Respir Res, 2010. 11: p. 170.
- Kim, W.H., et al., Expression of 32/67-kDa laminin receptor in laminin adhesion-selected human colon cancer cell lines. Br J Cancer, 1998. 77(1): p. 15-20.
- Iwamoto, Y., et al., YIGSR, a synthetic laminin pentapeptide, inhibits experimental metastasis formation. Science, 1987. 238(4830): p. 1132-4.
- Nomizu, M., et al., Multimeric forms of Tyr-Ile-Gly-Ser-Arg (YIGSR) peptide enhance the inhibition of tumor growth and metastasis. Cancer Res, 1993. 53(15): p. 3459-61.
- Yoshida, N., et al., The laminin-derived peptide YIGSR (Tyr-Ile-Gly-Ser-Arg) inhibits human pre-B leukaemic cell growth and dissemination to organs in SCID mice. Br J Cancer, 1999. 80(12): p. 1898-904.
- Yudoh, K., et al., Tumor cell attachment to laminin promotes degradation of the extracellular matrix and cell migration in high-metastatic clone cells of RCT sarcoma in vitro. Jpn J Cancer Res, 1995. 86(7): p. 685-90.
- Kim, W.H., et al., Apoptosis in human fibrosarcoma cells is induced by a multimeric synthetic Tyr-Ile-Gly-Ser-Arg (YIGSR)-containing polypeptide from laminin. Cancer Res, 1994. 54(18): p. 5005-10.
- Mu, Y., et al., Bioconjugation of laminin-related peptide YIGSR with polyvinyl pyrrolidone increases its antimetastatic effect due to a longer plasma half-life. Biochem Biophys Res Commun, 1999. 264(3): p. 763-7.
- Zhu, L., et al., Noncovalent Bonding of RGD and YIGSR to an Electrospun Poly(epsilon-Caprolactone) Conduit through Peptide Self-Assembly to Synergistically Promote Sciatic Nerve Regeneration in Rats. Adv Healthc Mater, 2017. 6(8).
- Wu, Z.Z., et al., Inhibition of adhesion of hepatocellular carcinoma cells to basement membrane components by receptor competition with RGD- or YIGSR-containing synthetic peptides. Biorheology, 2003. 40(4): p. 489-502.
- Lam, J., et al., Hydrogel design of experiments methodology to optimize hydrogel for iPSC-NPC culture. Adv Healthc Mater, 2015. 4(4): p. 534-9.
- Motta, C.M.M., et al., Enhancing Schwann cell migration using concentration gradients of laminin-derived peptides. Biomaterials, 2019. 218: p. 119335.
- Fittkau, M.H., et al., The selective modulation of endothelial cell mobility on RGD peptide containing surfaces by YIGSR peptides. Biomaterials, 2005. 26(2): p. 167-74.
- Wang, P.Y., et al., Grooved PLGA films incorporated with RGD/YIGSR peptides for potential application on skeletal muscle tissue engineering. Colloids Surf B Biointerfaces, 2013. 110: p. 88-95.
- Zustiak, S.P., R. Durbal, and J.B. Leach, Influence of cell-adhesive peptide ligands on poly(ethylene glycol) hydrogel physical, mechanical and transport properties. Acta Biomater, 2010. 6(9): p. 3404-14.