Here is a selection of publications where different laminin isoforms are being used to create more authentic cell culture systems
Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer
Berg K., Lange T., Mittelberger F., Schumacher U., Hahn U.Molecular Therapy—Nucleic Acids, 2016
Cancer cells use the α6β4 integrin/laminin-332 interaction to activate signaling pathways promoting tumor cell growth, invasion and metastasis, the inhibition of this interaction is of high therapeutic interest. Here, the authors report on the selection of a DNA aptamer inhibiting the interaction between α6β4 integrin and laminin-332. This Integrin α6β4-specific DNA Aptamer inhibits the adhesion of prostate cancer cells (PC-3) to laminin-332 with an IC50 value of 149 nmol/l. the aptamer was internalized into PC- 3-cells. Further characterization showed specificity to α6 integrins and a half-life in the murine blood plasma of 6 hours.
Monoclonal antibodies to human laminin α4 chain globular domain inhibit tumor cell adhesion and migration on laminins 411 and 421, and binding of α6β1 integrin and MCAM to α4-laminins
Ishikawa T., Wondimu Z., Oikawa Y., Ingerpuu S., Virtanen I., Patarroyo M.Matrix Biology, 2014
α4-Laminins, such as laminins -411 and -421, are mesenchymal laminins expressed by vascular and lymphatic endothelial cells, leukocytes and other normal cell types. These laminins are recognized by α6β1 and α6β4 integrins and MCAM (CD146) and promote adhesion and migration of the cells. α4-Laminins are also expressed and secreted by some tumor cells and strongly promote tumor cell migration. Moreover, the abluminal side of blood and/or lymphatic vessels and the nerve perineurium, common tracks of tumor cell dissemination, express α4-laminins, and these laminin isoforms, when expressed in the stroma, may contribute to tumor invasion. In the present study, we examined ten mAbs to human laminin α4 chain for their reactivity with the isolated laminin α4 globular domain, their ability to inhibit tumor cell adhesion and migration on laminin-411 and -421, and their effect on the binding of α6β1 integrin and MCAM to both α4-laminins. The results indicate that mAbs to the laminin α4 globular domain are able to inhibit tumor cell adhesion and migration on laminin-411 and -421 and that α6β1 integrin and MCAM bind α4-laminins at very close sites on the globular domain. These reagents contribute to a better understanding of the biology of α4-laminins and may have a therapeutic potential in malignant and inflammatory diseases.
Laminin-332 sustains chemoresistance and quiescence as part of the human hepatic cancer stem cell niche
Govaere O., Wouters J., Petz M., Vandewynckel Y-P., Van den Eynde K., Van den broeck A., Verhulst S., Dollé L., Gremeaux L., Ceulemans A., Nevens F., van Grunsven L.A., Topal B., Vankelecom H., Giannelli G., Van Vlierberghe H. Mikulits W., Komuta M., Roskams T.Journal of hepatology, 2015
This study demonstrates that tumor behavior is plastic and depends on the microenvironment of the tumor cell. We particularly identified an important role for laminin-332 and more specifically its gamma2-chain as part of the specialized cancer stem cell niche in maintaining and supporting ‘stemness’, e.g. quiescence and chemo-resistance. laminin-332 induces K19 expression, quiescence, and chemo-resistance in vitro. Laminin-332 not only protects hepatic cancer cells against chemotherapy but stimulates cell proliferation upon sorafenib exposure. Therefore, monoclonal antibody treatment targeting the gamma2-chain of laminin-332 could provide an innovative therapy of hepatic cancer.
Ultraviolet-radiation-induced inflammation promotes angiotropism and metastasis in melanoma
Bald T., Quast T., Landsberg J., Rogava M., Glodde n., Lopez-Ramos D., Kohlmeyer J., Riesenberg S., van den Boorn-Konijnenberg D., Hömig-Hölzel C., Reuten R., Schadow B., Weighardt H., Wenzel D., Helfrich I., Schadendorf D., Bloch W., Bianchi M.E., Lugassy C., Barnhill R.L., Koch M., Fleischmann B.K., Förster I., Kastenmüller W., Kolanus W., Hölzel M., Gaffal E., Tüting T. Nature Letter. 2014
The authors report that repetitive UV exposure of primary cutaneous melanomas in a genetically engineered mouse model promotes metastatic progression. UV irradiation enhanced the expansion of tumor cells along abluminal blood vessel surfaces and increased the number of lung metastases, depended on the recruitment and activation of neutrophils. In a static cell adhesion assays, cells were allowed to adhere to various matrices: fibronectin-1, collagen type I, collagen type IV, laminin-111 (Sigma), laminin-411 or laminin-511. An inflammatory environment promotes the ability of mouse and human melanoma cells to migrate towards endothelial cells and use selective motility cues on their surfaces.
Laminin Interactions with Head and Neck Cancer Cells under Low Fluid Shear Conditions Lead to Integrin Activation and Binding
Fennewald S.M., Kantara C., Sastry S.K., Resto V.AJournal of biological chemistry, 2012
Lymphatic metastasis of cancer cells involves movement from the primary tumor site to the lymph node, where the cells must be able to productively lodge and grow. Head and neck squamous cell carcinoma (HNSCC) cell lines cultured on placental laminin (laminin-511 is the major laminin), laminin-332 purified from human foreskin keratinocytes and human recombinant laminin-511, -211, -111, and -411. HNSCC cell lines bound to laminin-511 and -211 but also to -411 to a lower extent, under lymphodynamic low shear stress (0.07 dynes/cm2), consistent with lymph flow. Binding only occurred in the presence of shear stress and not in the absence of flow. The authors conclude that B1 integrins mediate tumor cell/lymph node interactions active under lymphodynamic flow. These interactions may drive growth and immunomodulation in this niche.
Gelatine methacrylamide-based hydrogels – an alternative 3D cancer cell culture system
Kaemmerer E., Melchels F.P.W, Holzapfel B.M, Meckel T., Hutmacher D.W., Loessner D. Acta Biomaterialia, 2014
The authors present a 3D biomaterial platform for the analysis of ovarian cancer spheroid growth that is an efficient semi-synthetic alternative, combining native ECM components and tunable matrix properties, resulting in higher reproducibility, less complexity and better comparability between different groups than traditional cell monolayer approaches. In this study, gelatine methacrylamide-based hydrogels (GelMA) with added LN-411 were established as in vitro and in vivo spheroid-based 3D cancer models.
Laminins and cancer stem cells: Partners in crime?
Qin Y., Rodin S., Simonson O.E., Hollande F. Seminars in Cancer Biology, 2016
A review that discusses the role of laminin as a regulator of cancer stem cells, in tumor progression and drug resistance. A growing body of literature suggests that laminins may act as regulators of cancer stem cells, a tumor cell subpopulation that plays an instrumental role in long-term cancer maintenance, metastasis development, and therapeutic resistance. The accumulating evidence in this emerging research area suggests that laminins represent potential therapeutic targets for anti-cancer treatments against cancer stem cells and that they may be used as predictive and prognostic markers to inform clinical management and improve patient survival.
Laminin 521 enhances self-renewal via STAT3 activation and promotes tumor progression in colorectal cancer
Qin Y, Shembrey C, Smith J, Paquet-Fifield S, Behrenbruch C, Beyit LM, Thomson BNJ, Heriot AG, Cao Y, Hollande F.Cancer Lett. 2020
Remodeling of basement membrane proteins contributes to tumor progression towards the metastatic stage. Here, the authors show that one of these proteins, laminin 521 (LN521), promotes colorectal cancer (CRC) cell self-renewal and invasion. siRNA-mediated knockdown of endogenously-produced laminin alpha 5, as well as treatment with neutralizing antibodies against integrin α3β1 and α6β1, were able to reverse the effect of LN521 on self-renewal. Exposure of CRC cells to LN521 enhanced STAT3 phosphorylation, and incubation with STAT3 inhibitors Napabucasin and Stattic were sufficient to block the LN521-driven self-renewal increase. Robust expression of laminin alpha 5 was detected in 7/10 liver metastases tissue sections collected from CRC patients as well as in mouse liver metastasis xenografts, in most cases within areas expressing metastasis cancer stem cell markers such as c-KIT and CD44v6. Finally, retrospective analysis of multiple CRC datasets highlighted the significant association between high LN521 mRNA expression and poor clinical outcome in colorectal cancer patients. Collectively our results indicate that high Laminin 521 expression is a frequent feature of metastatic dissemination in CRC and that it promotes cell invasion and self-renewal, the latter through the engagement of integrin isoforms and activation of STAT3 signaling.
L1CAM defines the regenerative origin of metastasis-initiating cells in colorectal cancer
Ganesh K., Basnet H., Kaygusuz Y., Laughney A.M., He L., Sharma R., O’Rourke K.P., Reuter V.P., Huang Y.-H., Turkekul M., Er E.E., Masilionis I., Manova-Todorova K., Weiser M.R., Saltz L.B., Garcia-Aguilar J., Koche R., Lowe S.W., Pe’er D., Shia J., Massagué J.Nature Cancer, 2020
The authors show that L1CAM+ cells in human colorectal cancer (CRC) have metastasis-initiating capacity, and they define their relationship to tissue regeneration. By using recombinant L1CAM extracellular domain and basement membrane components, they confirmed that L1CAM bound heterophilically to laminins known to be expressed in the intestinal and endothelial cell basement membranes in addition to exhibiting homophilic interaction with L1CAM itself. L1CAM knockdown inhibited the ability of CRC organoid-derived cells to bind to laminin-coated plates. Together, these data suggest that L1CAM enables the adhesion of metastasis-initiating cells to laminin-rich basement membranes, which is required for metastasis and organoid growth.
Laminin-421 produced by lymphatic endothelial cells induces chemotaxis for human melanoma cells
Saito N., Hamada J., Furukawa H., Tsutsumida A., Oyama A., Funayama E., Saito A., Tsuji T., Tada M., Moriuchi T., Yamamoto Y.Pigment Cell Melanoma Res., 2009
Here, the authors investigate the molecular mechanism of lymphatic metastasis. They examined the influence of interactions between normal lymphatic endothelial cells (LECs) and melanoma cells on cell migration. LEC conditioned medium (LEC-CM) contained chemotactic and chemokinetic activities for human melanoma cell lines. The chemotactic activity of LEC-CM was abolished by immunodepletion with anti-laminin-1 antibody. And immunoprecipitation and Western blot analyses revealed that LEC-CM contained laminin a4 and 5, b1 and 2, and c1, corresponding to isoforms -521, -511, -421 and -411. When melanoma C8161 cells were treated with function-blocking antibodies to integrin a3 or a6, their chemotactic responses to LEC-CM were markedly reduced. Furthermore, the knock-down of tetraspanin CD151 weakened the chemotactic responses of C8161 and MeWo cells to LEC-CM. These data suggest that laminin secreted by LEC possibly facilitates lymphatic metastasis through the induction of chemotaxis of melanoma cells.