276 results on '"Jyrki Heino"'
Search Results
2. Embigin deficiency leads to delayed embryonic lung development and high neonatal mortality in mice
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Salli Talvi, Johanna Jokinen, Kalle Sipilä, Pekka Rappu, Fu-Ping Zhang, Matti Poutanen, Pia Rantakari, and Jyrki Heino
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Developmental genetics ,Developmental biology ,Transcriptomics ,Science - Abstract
Summary: Embigin (Gp70), a receptor for fibronectin and an ancillary protein for monocarboxylate transporters, is known to regulate stem cell niches in sebaceous gland and bone marrow. Here, we show that embigin expression is at high level during early mouse embryogenesis and that embigin is essential for lung development. Markedly increased neonatal mortality of Emb−/− mice can be explained by the compromised lung maturation: in Emb−/− mice (E17.5) the number and the size of the small airways and distal airspace are significantly smaller, there are fewer ATI and ATII cells, and the alkaline phosphatase activity in amniotic fluid is lower. Emb−/− lungs show less peripheral branching already at E12.5, and embigin is highly expressed in lung primordium. Thus, embigin function is essential at early pseudoglandular stage or even earlier. Furthermore, our RNA-seq analysis and Ki67 staining results support the idea that the development of Emb−/− lungs is rather delayed than defected.
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- 2024
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3. Inflammation-related citrullination of matrisome proteins in human cancer
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Pekka Rappu, Ujjwal Suwal, Elina Siljamäki, and Jyrki Heino
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citrullination ,inflammation ,extracellular matrix ,cancer ,proteomics ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
IntroductionProtein arginine deiminases (PADs) are intracellular enzymes that may, especially in pathological conditions, also citrullinate extracellular substrates, including matrisome proteins such as structural proteins in extracellular matrix (ECM). PADs are abundantly expressed in human cancer cells. Citrullination of matrisome proteins has been reported in colon cancer but the phenomenon has never been systematically studied.MethodsTo gain a broader view of citrullination of matrisome proteins in cancer, we analyzed cancer proteomics data sets in 3 public databases for citrullinated matrisome proteins. In addition, we used three-dimensional cell cocultures of fibroblasts and cancer cells and analyzed citrullination of ECM.Results and discussionOur new analysis indicate that citrullination of ECM occurs in human cancer, and there is a significant variation between tumors. Most frequently citrullinated proteins included fibrinogen and fibronectin, which are typically citrullinated in rheumatoid inflammation. We also detected correlation between immune cell marker proteins, matrix metalloproteinases and ECM citrullination, which suggests that in cancer, citrullination of matrisome proteins is predominantly an inflammation-related phenomenon. This was further supported by our analysis of three-dimensional spheroid co-cultures of nine human cancer cell lines and fibroblasts by mass spectrometry, which gave no evidence that cancer cells or fibroblasts could citrullinate matrisome proteins in tumor stroma. It also appears that in the spheroid cultures, matrisome proteins are protected from citrullination.
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- 2022
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4. Unravelling the proteomic landscape of extracellular vesicles in prostate cancer by density-based fractionation of urine
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Bert Dhondt, Edward Geeurickx, Joeri Tulkens, Jan Van Deun, Glenn Vergauwen, Lien Lippens, Ilkka Miinalainen, Pekka Rappu, Jyrki Heino, Piet Ost, Nicolaas Lumen, Olivier De Wever, and An Hendrix
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extracellular vesicles ,exosomes ,separation ,isolation ,urine ,cancer ,biomarkers ,proteomics ,Cytology ,QH573-671 - Abstract
Extracellular vesicles (EV) are increasingly being recognized as important vehicles of intercellular communication and promising diagnostic and prognostic biomarkers in cancer. Despite this enormous clinical potential, the plethora of methods to separate EV from biofluids, providing material of highly variable purity, and lacking knowledge regarding methodological repeatability pose a barrier to clinical translation. Urine is considered an ideal proximal fluid for the study of EV in urological cancers due to its direct contact with the urogenital system. We demonstrate that density-based fractionation of urine by bottom-up Optiprep density gradient centrifugation separates EV and soluble proteins with high specificity and repeatability. Mass spectrometry-based proteomic analysis of urinary EV (uEV) in men with benign and malignant prostate disease allowed us to significantly expand the known human uEV proteome with high specificity and identifies a unique biological profile in prostate cancer not uncovered by the analysis of soluble proteins. In addition, profiling the proteome of EV separated from prostate tumour conditioned medium and matched uEV confirms the specificity of the identified uEV proteome for prostate cancer. Finally, a comparative proteomic analysis with uEV from patients with bladder and renal cancer provided additional evidence of the selective enrichment of protein signatures in uEV reflecting their respective cancer tissues of origin. In conclusion, this study identifies hundreds of previously undetected proteins in uEV of prostate cancer patients and provides a powerful toolbox to map uEV content and contaminants ultimately allowing biomarker discovery in urological cancers.
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- 2020
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5. Joint inflammation related citrullination of functional arginines in extracellular proteins
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Kalle H. Sipilä, Vipin Ranga, Pekka Rappu, Markku Mali, Laura Pirilä, Ilona Heino, Johanna Jokinen, Jarmo Käpylä, Mark S. Johnson, and Jyrki Heino
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Medicine ,Science - Abstract
Abstract We report the extent, specific sites and structural requirements of joint inflammation related citrullination in extracellular proteins. A total of 40 synovial fluid samples derived from chronically inflamed human joints were analysed by heparin-agarose fractionation and LC-MS/MS. Citrullination of 55 arginines in extracellular proteins was detected. Importantly, 20% of the sites have a characterized function related to the hallmarks of destructive joint inflammation. E.g. four arginine residues, shown here to be citrullinated, are also affected by mutations in inherited diseases causing haemolysis or blood clotting dysfunction. Citrullination of integrin ligands was selected for further studies since fibronectin R234 in isoDGR was among the most frequently citrullinated arginines in synovial fluid. Assays with synovial fibroblasts and integrin αVβ3 indicated decreased affinity to the enzymatically citrullinated integrin binding sites. To conclude, our data indicate that in inflamed joints extensive citrullination affects the functional arginine residues in extracellular proteins.
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- 2017
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6. Integrin α2β1 in nonactivated conformation can induce focal adhesion kinase signaling
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Maria Salmela, Johanna Jokinen, Silja Tiitta, Pekka Rappu, R. Holland Cheng, and Jyrki Heino
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Medicine ,Science - Abstract
Abstract Conformational activation of integrins is generally required for ligand binding and cellular signalling. However, we have previously reported that the nonactivated conformation of α2β1 integrin can also bind to large ligands, such as human echovirus 1. In this study, we show that the interaction between the nonactivated integrin and a ligand resulted in the activation of focal adhesion kinase (FAK) in a protein kinase C dependent manner. A loss-of-function mutation, α2E336A, in the α2-integrin did not prevent the activation of FAK, nor did EDTA-mediated inactivation of the integrin. Full FAK activation was observed, since phosphorylation was not only confirmed in residue Y397, but also in residues Y576/7. Furthermore, initiation of downstream signaling by paxillin phosphorylation in residue Y118 was evident, even though this activation was transient by nature, probably due to the lack of talin involvement in FAK activation and the absence of vinculin in the adhesion complexes formed by the nonactivated integrins. Altogether these results indicate that the nonactivated integrins can induce cellular signaling, but the outcome of the signaling differs from conventional integrin signaling.
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- 2017
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7. FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy
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Carlos J Diaz Osterman, Duygu Ozmadenci, Elizabeth G Kleinschmidt, Kristin N Taylor, Allison M Barrie, Shulin Jiang, Lisa M Bean, Florian J Sulzmaier, Christine Jean, Isabelle Tancioni, Kristen Anderson, Sean Uryu, Edward A Cordasco, Jian Li, Xiao Lei Chen, Guo Fu, Marjaana Ojalill, Pekka Rappu, Jyrki Heino, Adam M Mark, Guorong Xu, Kathleen M Fisch, Vihren N Kolev, David T Weaver, Jonathan A Pachter, Balázs Győrffy, Michael T McHale, Denise C Connolly, Alfredo Molinolo, Dwayne G Stupack, and David D Schlaepfer
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beta-catenin ,focal adhesion kinase FAK ,ovarian cancer ,platinum chemotherapy ,pluripotency ,tumorspheres ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Gene copy number alterations, tumor cell stemness, and the development of platinum chemotherapy resistance contribute to high-grade serous ovarian cancer (HGSOC) recurrence. Stem phenotypes involving Wnt-β-catenin, aldehyde dehydrogenase activities, intrinsic platinum resistance, and tumorsphere formation are here associated with spontaneous gains in Kras, Myc and FAK (KMF) genes in a new aggressive murine model of ovarian cancer. Adhesion-independent FAK signaling sustained KMF and human tumorsphere proliferation as well as resistance to cisplatin cytotoxicity. Platinum-resistant tumorspheres can acquire a dependence on FAK for growth. Accordingly, increased FAK tyrosine phosphorylation was observed within HGSOC patient tumors surviving neo-adjuvant chemotherapy. Combining a FAK inhibitor with platinum overcame chemoresistance and triggered cell apoptosis. FAK transcriptomic analyses across knockout and reconstituted cells identified 135 targets, elevated in HGSOC, that were regulated by FAK activity and β-catenin including Myc, pluripotency and DNA repair genes. These studies reveal an oncogenic FAK signaling role supporting chemoresistance.
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- 2019
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8. Long non-coding RNA PICSAR decreases adhesion and promotes migration of squamous carcinoma cells by downregulating α2β1 and α5β1 integrin expression
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Minna Piipponen, Jyrki Heino, Veli-Matti Kähäri, and Liisa Nissinen
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Long non-coding RNA ,PICSAR ,Integrin ,Adhesion ,Cutaneous squamous cell carcinoma ,Cancer ,Science ,Biology (General) ,QH301-705.5 - Abstract
Long non-coding RNAs (lncRNAs) regulate various cellular processes, and they have emerged as potential biomarkers and therapeutic targets in cancer. We have previously characterized the oncogenic role of lncRNA PICSAR (p38 inhibited cutaneous squamous cell carcinoma associated lincRNA) in cutaneous squamous cell carcinoma (cSCC), the most common metastatic skin cancer. In this study, we show that knockdown of PICSAR in cSCC cells upregulates expression of α2, α5 and β1 integrins, resulting in increased cell adhesion and decreased cell migration on collagen I and fibronectin. In contrast, overexpression of PICSAR in cSCC cells downregulates expression of α2, α5 and β1 integrins on cell surface, resulting in decreased cell adhesion on collagen I and fibronectin and increased cell migration. These results demonstrate a novel mechanism for regulation of the expression of collagen and fibronectin binding integrins by lncRNA PICSAR, leading to altered adhesion and migration of cSCC cells. This article has an associated First Person interview with the first author of the paper.
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- 2018
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9. Feasibility of Mechanical Extrusion to Coat Nanoparticles with Extracellular Vesicle Membranes
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Jan Van Deun, Quentin Roux, Sarah Deville, Thibaut Van Acker, Pekka Rappu, Ilkka Miinalainen, Jyrki Heino, Frank Vanhaecke, Bruno G. De Geest, Olivier De Wever, and An Hendrix
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extracellular vesicle ,exosome ,microvesicle ,biomimetics ,gold nanoparticle ,functionalization ,Cytology ,QH573-671 - Abstract
Biomimetic functionalization to confer stealth and targeting properties to nanoparticles is a field of intense study. Extracellular vesicles (EV), sub-micron delivery vehicles for intercellular communication, have unique characteristics for drug delivery. We investigated the top-down functionalization of gold nanoparticles with extracellular vesicle membranes, including both lipids and associated membrane proteins, through mechanical extrusion. EV surface-exposed membrane proteins were confirmed to help avoid unwanted elimination by macrophages, while improving autologous uptake. EV membrane morphology, protein composition and orientation were found to be unaffected by mechanical extrusion. We implemented complementary EV characterization methods, including transmission- and immune-electron microscopy, and nanoparticle tracking analysis, to verify membrane coating, size and zeta potential of the EV membrane-cloaked nanoparticles. While successful EV membrane coating of the gold nanoparticles resulted in lower macrophage uptake, low yield was found to be a significant downside of the extrusion approach. Our data incentivize more research to leverage EV membrane biomimicking as a unique drug delivery approach in the near future.
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- 2020
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10. Early chordate origin of the vertebrate integrin αI domains.
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Bhanupratap Singh Chouhan, Jarmo Käpylä, Konstantin Denessiouk, Alexander Denesyuk, Jyrki Heino, and Mark S Johnson
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Medicine ,Science - Abstract
Half of the 18 human integrins α subunits have an inserted αI domain yet none have been observed in species that have diverged prior to the appearance of the urochordates (ascidians). The urochordate integrin αI domains are not human orthologues but paralogues, but orthologues of human αI domains extend throughout later-diverging vertebrates and are observed in the bony fish with duplicate isoforms. Here, we report evidence for orthologues of human integrins with αI domains in the agnathostomes (jawless vertebrates) and later diverging species. Sequence comparisons, phylogenetic analyses and molecular modeling show that one nearly full-length sequence from lamprey and two additional fragments include the entire integrin αI domain region, have the hallmarks of collagen-binding integrin αI domains, and we show that the corresponding recombinant proteins recognize the collagen GFOGER motifs in a metal dependent manner, unlike the α1I domain of the ascidian C. intestinalis. The presence of a functional collagen receptor integrin αI domain supports the origin of orthologues of the human integrins with αI domains prior to the earliest diverging extant vertebrates, a domain that has been conserved and diversified throughout the vertebrate lineage.
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- 2014
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11. Melanoma-associated cancer-testis antigen 16 (CT16) regulates the expression of apoptotic and antiapoptotic genes and promotes cell survival.
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Camilla Nylund, Pekka Rappu, Eveliina Pakula, Aleksi Heino, Laura Laato, Laura L Elo, Pia Vihinen, Seppo Pyrhönen, Gethin R Owen, Hannu Larjava, Markku Kallajoki, and Jyrki Heino
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Medicine ,Science - Abstract
Cancer-testis (CT) antigens are predominantly expressed in testis or placenta, but absent in most adult tissues. During malignant transformation CT genes are often activated. CT antigen 16 (CT16, PAGE5) is frequently expressed in advanced melanoma but its biological function has been unknown. To examine the role of CT16 in cell survival we knocked it down in A2058 melanoma cells using specific siRNAs and exposed the cells to cancer drug cisplatin known to induce apoptosis. As a result, cell survival was markedly decreased. To study the effects of CT16 on cell survival in more detail, the cellular gene expression profiles were investigated after CT16 silencing in CT16 positive A2058 melanoma cells, as well as after CT16 overexpression in CT16 negative WM-266-4 melanoma cells. Among the 11 genes both upregulated by CT16 silencing and downregulated by CT16 overexpression or vice versa, 4 genes were potentially apoptotic or antiapoptotic genes. CT16 was recognized as a positive regulator of antiapoptotic metallothionein 2A and interleukin 8 genes, whereas it inhibited the expression of apoptosis inducing dickkopf 1 (DKK1) gene. In addition CT16 enhanced the expression of fatty acid binding protein 7, a known promoter of melanoma progression. The effect of CT16 on DKK1 expression was p53 independent. Furthermore, CT16 did not regulate apoptotic genes via DNA methylation. In twenty melanoma metastasis tissue samples average DKK1 mRNA level was shown to be significantly (p
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- 2012
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12. Conservation of the human integrin-type beta-propeller domain in bacteria.
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Bhanupratap Chouhan, Alexander Denesyuk, Jyrki Heino, Mark S Johnson, and Konstantin Denessiouk
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Medicine ,Science - Abstract
Integrins are heterodimeric cell-surface receptors with key functions in cell-cell and cell-matrix adhesion. Integrin α and β subunits are present throughout the metazoans, but it is unclear whether the subunits predate the origin of multicellular organisms. Several component domains have been detected in bacteria, one of which, a specific 7-bladed β-propeller domain, is a unique feature of the integrin α subunits. Here, we describe a structure-derived motif, which incorporates key features of each blade from the X-ray structures of human αIIbβ3 and αVβ3, includes elements of the FG-GAP/Cage and Ca(2+)-binding motifs, and is specific only for the metazoan integrin domains. Separately, we searched for the metazoan integrin type β-propeller domains among all available sequences from bacteria and unicellular eukaryotic organisms, which must incorporate seven repeats, corresponding to the seven blades of the β-propeller domain, and so that the newly found structure-derived motif would exist in every repeat. As the result, among 47 available genomes of unicellular eukaryotes we could not find a single instance of seven repeats with the motif. Several sequences contained three repeats, a predicted transmembrane segment, and a short cytoplasmic motif associated with some integrins, but otherwise differ from the metazoan integrin α subunits. Among the available bacterial sequences, we found five examples containing seven sequential metazoan integrin-specific motifs within the seven repeats. The motifs differ in having one Ca(2+)-binding site per repeat, whereas metazoan integrins have three or four sites. The bacterial sequences are more conserved in terms of motif conservation and loop length, suggesting that the structure is more regular and compact than those example structures from human integrins. Although the bacterial examples are not full-length integrins, the full-length metazoan-type 7-bladed β-propeller domains are present, and sometimes two tandem copies are found.
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- 2011
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13. Characterization of intrinsically disordered prostate associated gene (PAGE5) at single residue resolution by NMR spectroscopy.
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Maarit Hellman, Helena Tossavainen, Pekka Rappu, Jyrki Heino, and Perttu Permi
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Medicine ,Science - Abstract
BACKGROUND: The Cancer-Testis antigens (CTA) are proteins expressed in human germ line and certain cancer cells. CTAs form a large gene family, representing 10% of X-chromosomal genes. They have high potential for cancer-specific immunotherapy. However, their biological functions are currently unknown. Prostate associated genes (PAGE) are characterized as CTAs. PAGE5 is one of six proteins belonging to this protein family, also called CT16. METHODOLOGY/PRINCIPAL FINDINGS: In this study we show, using bioinformatics, chromatographic and solution state NMR spectroscopic methods, that PAGE5 is an intrinsically disordered protein (IDP). CONCLUSION/SIGNIFICANCE: The study stands out as the first time structural characterization of the PAGE family protein and introduces how solution state NMR spectroscopy can be effectively utilized for identification of molecular recognition regions (MoRF) in IDPs, known often as transiently populated secondary structures.
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- 2011
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14. What does it take to be a collagen receptor?
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Richard Farndale, Arnoud Sonnenberg, C. Michael DiPersio, Johannes A Eble, Jyrki Heino, and Donald Gullberg
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Molecular Biology - Published
- 2023
15. Embigin deficiency leads to delayed embryonic lung development and high neonatal mortality
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Salli Talvi, Matti Poutanen, Kalle Sipilä, Jyrki Heino, Pia Rantakari, Pekka Rappu, Johanna Jokinen, and Fu-Ping Zhang
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0303 health sciences ,Morphogenesis ,Hematopoietic stem cell ,Embryo ,Biology ,Embryonic stem cell ,3. Good health ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Placenta ,Gene expression ,medicine ,Progenitor cell ,Homeostasis ,030304 developmental biology - Abstract
Embigin (gp70), a transmembrane glycoprotein, has been shown to regulate hematopoietic stem cell and progenitor cell niche. Still, little is known about its expression and function in other organ systems during development or adulthood. By combining immunofluorescence, RNA sequencing, and in vivo mouse models, we show that embigin is highly expressed during development and in adult lung, kidney, epididymis, skin, and testis. Adult Emb-/- mice have a normal lifespan and fertility without apparent pathologies. In contrast, the Emb-/- embryos are significantly smaller than their WT littermates. Markedly increased mortality of the Emb-/- embryos is seen especially during the neonatal period. Embigin is present in the placenta, but placental morphology and gene expression patterns stay unaltered. At E17.5, Emb-/- mice show defective morphogenesis of the lung, low alkaline phosphatase activity in amniotic fluid, and remarkable activation of genes involved in cell proliferation in the lungs. Thus, lung underdevelopment explains the high neonatal mortality. Our work demonstrates the crucial role of embigin during development, and it paves the way to further characterization of embigin in specific organ systems in development and homeostasis.Summary statementEmbigin is a basigin-group transmembrane glycoprotein. In vivo mouse model shows that embigin is crucial for embryonic lung development and neonatal survival.
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- 2021
16. Embigin is a fibronectin receptor that affects sebaceous gland differentiation and metabolism
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Kalle Sipilä, Emanuel Rognoni, Johanna Jokinen, Mukul Tewary, Matteo Vietri Rudan, Salli Talvi, Ville Jokinen, Käthe M. Dahlström, Kif Liakath-Ali, Atefeh Mobasseri, Xinyi Du-Harpur, Jarmo Käpylä, Stephen L. Nutt, Tiina A. Salminen, Jyrki Heino, and Fiona M. Watt
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Integrins ,Mice ,Sebaceous Glands ,Integrin beta1 ,Cell Adhesion ,Animals ,Cell Differentiation ,Cell Biology ,Molecular Biology ,General Biochemistry, Genetics and Molecular Biology ,Fibronectins ,Integrin alpha5beta1 ,Developmental Biology - Abstract
Stem cell renewal and differentiation are regulated by interactions with the niche. Although multiple cell populations have been identified in distinct anatomical compartments, little is known about niche-specific molecular factors. Using skin as a model system and combining single-cell RNA-seq data analysis, immunofluorescence, and transgenic mouse models, we show that the transmembrane protein embigin is specifically expressed in the sebaceous gland and that the number of embigin-expressing cells is negatively regulated by Wnt. The loss of embigin promotes exit from the progenitor compartment and progression toward differentiation, and also compromises lipid metabolism. Embigin modulates sebaceous niche architecture by affecting extracellular matrix organization and basolateral targeting of monocarboxylate transport. We discover through ligand screening that embigin is a direct fibronectin receptor, binding to the N-terminal fibronectin domain without impairing integrin function. Our results solve the long-standing question of how embigin regulates cell adhesion and demonstrate a mechanism that couples adhesion and metabolism.
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- 2022
17. C1r Upregulates Production of Matrix Metalloproteinase-13 and Promotes Invasion of Cutaneous Squamous Cell Carcinoma
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Liisa Nissinen, Marjaana Ojalill, Markku Kallajoki, Pilvi Riihilä, Jyrki Heino, Kristina Viiklepp, Veli-Matti Kähäri, Seppo Meri, HUSLAB, Seppo Meri / Principal Investigator, Department of Bacteriology and Immunology, University of Helsinki, and TRIMM - Translational Immunology Research Program
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EXPRESSION ,Skin Neoplasms ,MMP1 ,MMP10 ,Extracellular matrix component ,PROGRESSION ,Dermatology ,Complement factor I ,Matrix metalloproteinase ,Biochemistry ,COMPLEMENT ,HETEROGENEOUS DISORDER ,03 medical and health sciences ,Classical complement pathway ,0302 clinical medicine ,Cell Movement ,Cell Line, Tumor ,Matrix Metalloproteinase 13 ,Animals ,Humans ,Molecular Biology ,Cell Proliferation ,030304 developmental biology ,0303 health sciences ,Complement C1r ,Chemistry ,PERIODONTAL-DISEASE ,Cell migration ,KERATINOCYTE ,Cell Biology ,3. Good health ,Gene Expression Regulation, Neoplastic ,COLLAGENASE-3 MMP-13 ,3121 General medicine, internal medicine and other clinical medicine ,030220 oncology & carcinogenesis ,Metalloendopeptidase activity ,Carcinoma, Squamous Cell ,Cancer research ,GROWTH ,Heterografts ,TUMOR MICROENVIRONMENT ,EHLERS-DANLOS-SYNDROME - Abstract
Cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer, with increasing incidence worldwide. Previous studies have shown the role of the complement system in cSCC progression. In this study, we have investigated the mechanistic role of serine proteinase C1r, a component of the classical pathway of the complement system, in cSCC. Knockout of C1r in cSCC cells using CRISPR/Cas9 resulted in a significant decrease in their proliferation, migration, and invasion through collagen type I compared with that of wild-type cSCC cells. Knockout of C1r suppressed the growth and vascularization of cSCC xenograft tumors and promoted apoptosis of tumor cells in vivo. mRNA-sequencing analysis after C1r knockdown revealed significantly regulated Gene Ontology terms cell-matrix adhesion, extracellular matrix component, basement membrane, and metalloendopeptidase activity and Kyoto Encyclopedia of Genes and Genomes pathway extracellular matrix-receptor interaction. Among the significantly regulated genes were invasion-associated matrix metalloproteinases (MMPs) MMP1, MMP13, MMP10, and MMP12. Knockout of C1r resulted in decreased production of MMP-1, MMP-13, MMP-10, and MMP-12 by cSCC cells in culture. Knockout of C1r inhibited the expression of MMP-13 by tumor cells, suppressed invasion, and reduced the amount of degraded collagen in vivo in xenografts. These results provide evidence for the role of C1r in promoting the invasion of cSCC cells by increasing MMP production.
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- 2022
18. Integrin α11β1 is a receptor for collagen XIII
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Jarkko Koivunen, Anne Heikkinen, Mikko A. J. Finnilä, Padmanabhan Anbazhagan, André H. Juffer, Hongmin Tu, Donald Gullberg, Jarmo Käpylä, Antti Kemppainen, Jyrki Heino, Ning Lu, Simo Saarakkala, and Taina Pihlajaniemi
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Integrins ,Receptors, Collagen ,Histology ,ECM receptors ,Integrin ,Collagen Type XIII ,Bone and Bones ,Cell Line ,Pathology and Forensic Medicine ,Collagen receptor ,Mice ,Hydroxyproline ,chemistry.chemical_compound ,Animals ,Humans ,Bone homeostasis ,Cell adhesion ,Receptor ,Mice, Knockout ,biology ,Mesenchymal stem cell ,Regular Article ,Cell Biology ,Transfection ,Cell biology ,chemistry ,biology.protein ,Collagen ,Integrin alpha Chains ,C2C12 - Abstract
Collagen XIII is a conserved transmembrane collagen mainly expressed in mesenchymal tissues. Previously, we have shown that collagen XIII modulates tissue development and homeostasis. Integrins are a family of receptors that mediate signals from the environment into the cells and vice versa. Integrin α11β1 is a collagen receptor known to recognize the GFOGER (O=hydroxyproline) sequence in collagens. Interestingly, collagen XIII and integrin α11β1 both have a role in the regulation of bone homeostasis. To study whether α11β1 is a receptor for collagen XIII, we utilized C2C12 cells transfected to express α11β1 as their only collagen receptor. The interaction between collagen XIII and integrin α11β1 was also confirmed by surface plasmon resonance and pull-down assays. We discovered that integrin α11β1 mediates cell adhesion to two collagenous motifs, namely GPKGER and GF(S)QGEK, that were shown to act as the recognition sites for the integrin α11-I domain. Furthermore, we studied the in vivo significance of the α11β1-collagen XIII interaction by crossbreeding α11 null mice (Itga11−/−) with mice overexpressing Col13a1 (Col13a1oe). When we evaluated the bone morphology by microcomputed tomography, Col13a1oe mice had a drastic bone overgrowth followed by severe osteoporosis, whereas the double mutant mouse line showed a much milder bone phenotype. To conclude, our data identifies integrin α11β1 as a new collagen XIII receptor and demonstrates that this ligand-receptor pair has a role in the maintenance of bone homeostasis.
- Published
- 2021
19. Interaction between prostate cancer cells and prostate fibroblasts promotes accumulation and proteolytic processing of basement membrane proteins
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Pekka Rappu, Elina Siljamäki, Marjaana Ojalill, Jyrki Heino, Pekka Taimen, and Noora Virtanen
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0301 basic medicine ,Male ,Proteomics ,Urology ,Cathepsin L ,Cell ,Cell Communication ,Autoantigens ,Basement Membrane ,Mass Spectrometry ,Extracellular matrix ,03 medical and health sciences ,0302 clinical medicine ,DU145 ,Cancer-Associated Fibroblasts ,Cell Movement ,Cell Line, Tumor ,Spheroids, Cellular ,medicine ,Humans ,Neoplasm Invasiveness ,Fibroblast ,Basement membrane ,Tumor microenvironment ,Chemistry ,Membrane Proteins ,Prostatic Neoplasms ,Non-Fibrillar Collagens ,Cell biology ,Extracellular Matrix ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Cancer cell ,PC-3 Cells ,Endostatin ,Heparan Sulfate Proteoglycans - Abstract
Background Tumor microenvironment or stroma has the potency to regulate the behavior of malignant cells. Fibroblast-like cells are abundant in tumor stroma and they are also responsible for the synthesis of many extracellular matrix components. Fibroblast-cancer cell interplay can modify the functions of both cell types. Methods We applied mass spectrometry and proteomics to unveil the matrisome in 3D spheroids formed by DU145 prostate cancer cells, PC3 prostate cancer cells, or prostate-derived fibroblasts. Similarly, DU145/fibroblast and PC3/fibroblast coculture spheroids were also analyzed. Western blot analysis and immunofluorescence were used to confirm the presence of specific proteins in spheroids. Cancer dissemination was studied by utilizing "out of spheroids" migration and invasion assays. Results In the spheroid model cancer cell-fibroblast interplay caused remarkable changes in the extracellular matrix and accelerated the invasion of DU145 cells. Fibroblasts produced structural matrix proteins, growth factors, and matrix metalloproteinases. In cancer cell/fibroblast cocultures basement membrane components, including laminins (α3, α5, β2, and β3), heparan sulfate proteoglycan (HSPG2 gene product), and collagen XVIII accumulated in a prominent manner when compared with spheroids that contained fibroblasts or cancer cells only. Furthermore, collagen XVIII was intensively processed to different endostatin-containing isoforms by cancer cell-derived cathepsin L. Conclusions Fibroblasts can promote carcinoma cell dissemination by several different mechanisms. Extracellular matrix and basement membrane proteins provide attachment sites for cell locomotion promoting adhesion receptors. Growth factors and metalloproteinases are known to accelerate cell invasion. In addition, cancer cell-fibroblast interplay generates biologically active fragments of basement membrane proteins, such as endostatin.
- Published
- 2020
20. Integration of Matrisome Omics: Towards System Biology of the Tumor Matrisome
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Taina Pihlajaniemi, Valerio Izzi, Jyrki Heino, Pekka Rappu, and Jarkko Koivunen
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Tumor microenvironment ,Computer science ,business.industry ,Systems biology ,Big data ,Identification (biology) ,Computational biology ,business ,Proteomics ,Omics - Abstract
The tumor matrisome, the collection of structural and functional extracellular matrix (ECM) proteins found in tumors together with ECM-associated proteins, growth factors, cytokines and chemokines is an extremely dynamic entity whose composition and regulation depends on both local, cell-specific processes and overarching pan-cancer microenvironmental landscapes. A deeper knowledge and understanding of the tumor matrisome would grant us novel diagnostic, prognostic and therapeutic possibilities but to reach that point there is a pressing need to combine fine-grained molecular studies with system biology approaches to build a complete map of what actually happens in the tumor microenvironment. In this chapter, we will review the most salient findings on the tumor matrisome at the transcriptomics, regulomics and proteomics level and show how the use of big data enables the identification of biological features and characteristics that cross different tumors and the discovery of novel molecular mechanisms connecting the different omics level. Also, we will provide a simple walkthrough to jumpstart matrisome data analysis for complete beginners in a bid to ignite more research in this fascinating field.
- Published
- 2020
21. Unravelling the proteomic landscape of extracellular vesicles in prostate cancer by density-based fractionation of urine
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Piet Ost, Bert Dhondt, Nicolaas Lumen, Edward Geeurickx, An Hendrix, Joeri Tulkens, Jyrki Heino, Glenn Vergauwen, Olivier De Wever, Jan Van Deun, Lien Lippens, Pekka Rappu, and Ilkka Miinalainen
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0301 basic medicine ,EXPRESSION ,separation ,Histology ,BIOMARKERS ,PROTEIN ,exosomes ,Urine ,Proteomics ,Extracellular vesicles ,03 medical and health sciences ,Prostate cancer ,0302 clinical medicine ,proteomics ,Prostate ,medicine ,Medicine and Health Sciences ,cancer ,lcsh:QH573-671 ,Biomarker discovery ,MICROVESICLES ,EXOSOMES ,IDENTIFICATION ,lcsh:Cytology ,Chemistry ,biomarkers ,Cell Biology ,medicine.disease ,Microvesicles ,urine ,030104 developmental biology ,medicine.anatomical_structure ,DIFFERENTIATION ,030220 oncology & carcinogenesis ,Proteome ,Cancer research ,GROWTH ,extracellular vesicles ,isolation ,Research Article - Abstract
Extracellular vesicles (EV) are increasingly being recognized as important vehicles of intercellular communication and promising diagnostic and prognostic biomarkers in cancer. Despite this enormous clinical potential, the plethora of methods to separate EV from biofluids, providing material of highly variable purity, and lacking knowledge regarding methodological repeatability pose a barrier to clinical translation. Urine is considered an ideal proximal fluid for the study of EV in urological cancers due to its direct contact with the urogenital system. We demonstrate that density-based fractionation of urine by bottom-up Optiprep density gradient centrifugation separates EV and soluble proteins with high specificity and repeatability. Mass spectrometry-based proteomic analysis of urinary EV (uEV) in men with benign and malignant prostate disease allowed us to significantly expand the known human uEV proteome with high specificity and identifies a unique biological profile in prostate cancer not uncovered by the analysis of soluble proteins. In addition, profiling the proteome of EV separated from prostate tumour conditioned medium and matched uEV confirms the specificity of the identified uEV proteome for prostate cancer. Finally, a comparative proteomic analysis with uEV from patients with bladder and renal cancer provided additional evidence of the selective enrichment of protein signatures in uEV reflecting their respective cancer tissues of origin. In conclusion, this study identifies hundreds of previously undetected proteins in uEV of prostate cancer patients and provides a powerful toolbox to map uEV content and contaminants ultimately allowing biomarker discovery in urological cancers.
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- 2020
22. The binding capacity of α1β1-, α2β1- and α10β1-integrins depends on non-collagenous surface macromolecules rather than the collagens in cartilage fibrils
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Peter Bruckner, Mikko Huhtala, Uwe Hansen, Johannes A. Eble, Christian Gil Girol, Jarmo Käpylä, Jyrki Heino, Birgit Leitinger, Rita Dreier, Christian Woltersdorf, Stephan Niland, Melanie Bonk, and Biotechnology and Biological Sciences Research Council (BBSRC)
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Cartilage, Articular ,0301 basic medicine ,Biochemistry & Molecular Biology ,Fibrillar Collagens ,Integrin ,Chick Embryo ,Fibril ,Chondrocyte ,Integrin alpha1beta1 ,Extracellular matrix ,03 medical and health sciences ,Chondrocytes ,0302 clinical medicine ,Mechanoreception ,Adaptor proteins ,Cell Adhesion ,medicine ,Animals ,Humans ,Cell adhesion ,Discoidin Domain Receptors ,Molecular Biology ,Cells, Cultured ,Integrin binding ,Suprastructure ,biology ,Cell adhesion molecule ,Chemistry ,Cell-matrix-interactions ,Fibrillogenesis ,06 Biological Sciences ,Cell biology ,Immobilized Proteins ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,Cattle ,Integrin alpha2beta1 ,Integrin alpha Chains ,Protein Binding - Abstract
Interactions of cells with supramolecular aggregates of the extracellular matrix (ECM) are mediated, in part, by cell surface receptors of the integrin family. These are important molecular components of cell surface-suprastructures regulating cellular activities in general. A subfamily of β1-integrins with von Willebrand-factor A-like domains (I-domains) in their α-chains can bind to collagen molecules and, therefore, are considered as important cellular mechano-receptors. Here we show that chondrocytes strongly bind to cartilage collagens in the form of individual triple helical molecules but very weakly to fibrils formed by the same molecules. We also find that chondrocyte integrins α1β1-, α2β1- and α10β1-integrins and their I-domains have the same characteristics. Nevertheless we find integrin binding to mechanically generated cartilage fibril fragments, which also comprise peripheral non-collagenous material. We conclude that cell adhesion results from binding of integrin-containing adhesion suprastructures to the non-collagenous fibril periphery but not to the collagenous fibril cores. The biological importance of the well-investigated recognition of collagen molecules by integrins is unknown. Possible scenarios may include fibrillogenesis, fibril degradation and/or phagocytosis, recruitment of cells to remodeling sites, or molecular signaling across cytoplasmic membranes. In these circumstances, collagen molecules may lack a fibrillar organization. However, other processes requiring robust biomechanical functions, such as fibril organization in tissues, cell division, adhesion, or migration, do not involve direct integrin-collagen interactions.
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- 2017
23. Effect of micro-pores on cracks formation in metallurgical coke
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Timo Fabritius, Stanislav S. Gornostayev, and Jyrki Heino
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chemistry.chemical_classification ,metallurgical coke ,Materials science ,Thermoplastic ,Metallurgy ,Metals and Alloys ,pyrometallurgy ,Metallurgical coke ,02 engineering and technology ,Coke ,Industrial and Manufacturing Engineering ,020501 mining & metallurgy ,Matrix (geology) ,law.invention ,Field emission microscopy ,0205 materials engineering ,chemistry ,Resist ,Optical microscope ,pores ,law ,cracks - Abstract
The relationships of micro-pores and cracks in metallurgical coke have been investigated by optical microscope and field emission scanning electron microscope, using surface section samples. The pores have circular, elliptical and irregular shapes with smooth outlines, formed during the thermoplastic stage of the coking process. They often associate with connecting cracks between neighbouring pores. In case of elliptical pores, the connecting cracks are usually oriented along the longer axis of the pore. The connecting cracks can be developed between the pores, depending on their size and the distance between them. The coke with a large number of small pores rather than with a small number of larger pores will have lower strength due to the increased amount of connecting cracks. When compared with circular pores, elliptical and flattened pores have a lower ability to resist load pressure. Nano-sized pores have polygonal outlines, indicating an ‘explosion’-type formation in the solidified matrix. Résumé On a examiné la relation entre les micropores et les fissures du coke métallurgique au moyen de la microscopie optique et de la FESEM, en utilisant des échantillons de section de la surface. Les pores ont des formes circulaires, elliptiques ou irrégulières avec des contours lisses, et sont formés lors de l’étape thermoplastique du procédé de cokéfaction. Ils sont souvent associés à la mise en connexion des fissures entre les pores voisins. Dans le cas des pores elliptiques, les fissures de connexion sont habituellement orientées dans la direction de l’axe le plus long du pore. Les fissures de connexion peuvent se développer entre les pores, dépendant de leurs tailles et de la distance entre eux. Le coke ayant un grand nombre de petit pores plutôt qu’un petit nombre de pores plus grands aura une plus faible résistance à cause de la plus grande quantité de fissures de connexion. Lorsque comparés aux pores circulaires, les pores elliptiques et aplatis ont une habilité plus faible de résister à la pression de la charge. Les pores nanométriques ont des contours polygonaux, indiquant une formation de type ‘explosion’ dans la matrice solidifiée.
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- 2017
24. Complement component C1r upregulates the expression of matrix metalloproteinase-1, -13 and -10 and promotes invasion of cutaneous squamous carcinoma cells
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Veli-Matti Kähäri, Jyrki Heino, Seppo Meri, Pilvi Riihilä, Marjaana Ojalill, Liisa Nissinen, and Kristina Viiklepp
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- 2019
25. FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy
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Isabelle Tancioni, Duygu Ozmadenci, Sean Uryu, Michael T. McHale, K.M. Anderson, Guorong Xu, Dwayne G. Stupack, Vihren N. Kolev, Edward A. Cordasco, Denise C. Connolly, Jonathan A. Pachter, Jyrki Heino, Shulin Jiang, L.M. Bean, Kristin N. Taylor, Christine Jean, Balázs Győrffy, Elizabeth G. Kleinschmidt, Xiao Lei Chen, Jian Li, David D. Schlaepfer, A. Barrie, Alfredo A. Molinolo, David T. Weaver, Marjaana Ojalill, Adam Mark, Pekka Rappu, Carlos J. Díaz Osterman, Guo Fu, Florian J. Sulzmaier, and Kathleen M. Fisch
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0301 basic medicine ,Mouse ,endocrine system diseases ,medicine.medical_treatment ,Drug Resistance ,Disease ,medicine.disease_cause ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,Ovarian carcinoma ,2.1 Biological and endogenous factors ,Biology (General) ,Aetiology ,Cancer Biology ,cancer biology ,Cancer ,Ovarian Neoplasms ,0303 health sciences ,biology ,Stem Cells ,General Neuroscience ,General Medicine ,platinum chemotherapy ,female genital diseases and pregnancy complications ,Ovarian Cancer ,3. Good health ,ovarian cancer ,Paclitaxel ,030220 oncology & carcinogenesis ,Medicine ,Female ,KRAS ,Research Article ,Signal Transduction ,focal adhesion kinase FAK ,Beta-catenin ,tumorspheres ,QH301-705.5 ,DNA repair ,Science ,PTK2 ,Antineoplastic Agents ,General Biochemistry, Genetics and Molecular Biology ,Proto-Oncogene Proteins c-myc ,Proto-Oncogene Proteins p21(ras) ,03 medical and health sciences ,Rare Diseases ,Cancer stem cell ,Genetics ,medicine ,Animals ,Humans ,Gene ,mouse ,Platinum ,030304 developmental biology ,Chemotherapy ,General Immunology and Microbiology ,Animal ,business.industry ,beta-catenin ,pluripotency ,Stem Cell Research ,medicine.disease ,Disease Models, Animal ,030104 developmental biology ,chemistry ,Drug Resistance, Neoplasm ,Focal Adhesion Kinase 1 ,Catenin ,Disease Models ,Cancer cell ,biology.protein ,Cancer research ,Neoplasm ,Biochemistry and Cell Biology ,Cisplatin ,business ,Ovarian cancer ,DNA - Abstract
Gene copy number alterations, tumor cell stemness, and the development of platinum chemotherapy resistance contribute to high-grade serous ovarian cancer (HGSOC) recurrence. Stem phenotypes involving Wnt-β-catenin, aldehyde dehydrogenase activities, intrinsic platinum resistance, and tumorsphere formation are here associated with spontaneous gains in Kras, Myc and FAK (KMF) genes in a new aggressive murine model of ovarian cancer. Adhesion-independent FAK signaling sustained KMF and human tumorsphere proliferation as well as resistance to cisplatin cytotoxicity. Platinum-resistant tumorspheres can acquire a dependence on FAK for growth. Accordingly, increased FAK tyrosine phosphorylation was observed within HGSOC patient tumors surviving neo-adjuvant chemotherapy. Combining a FAK inhibitor with platinum overcame chemoresistance and triggered cell apoptosis. FAK transcriptomic analyses across knockout and reconstituted cells identified 135 targets, elevated in HGSOC, that were regulated by FAK activity and β-catenin including Myc, pluripotency and DNA repair genes. These studies reveal an oncogenic FAK signaling role supporting chemoresistance., eLife digest Ovarian cancer is one of the deadliest types of cancer in women. There are two main reasons for the aggressiveness of this cancer. First, ovarian cancer cells can spread to other parts of a woman’s body before she has been diagnosed, where the cells grow as tiny clumps or spheres of tumor cells, also called tumorspheres. Second, in the majority of patients, some ovarian cancer cells will develop resistance to the chemotherapy used. It is not clear exactly how these tumor cells become resistant to therapy. One way in which cells could do this is by gaining extra copies of genes that remove toxic substances or repair DNA, which help them withstand the therapy. Here, Osterman, Ozmadenci, Keinschmidt, Taylor, Barrie, Jiang, Bean, Sulzmaier et al. set up a new experimental method to study how some ovarian cancer cells resist chemotherapy. Comparing ovarian cancer cells from mice at early and late stages of the disease showed that the later-stage, more aggressive cells had more genetic changes. One of these changes affected the gene for a protein called FAK, which was found to have more copies than normal. The FAK protein is an enzyme that helps cancer cells move around. In cells from mice with late-stage cancer, FAK was over-active and present at high levels. When these cells grew as tumorspheres, the tumors were more resistant to chemotherapy than their early-stage counterparts. In patients who have received chemotherapy, surviving tumor cells also exhibit high levels of FAK activity. Human ovarian cancer cells that are resistant to chemotherapy can be grown into tumors in mice, where they retain their resistance to chemotherapy. However, if chemotherapy is combined with a drug that targets the FAK enzyme, the tumors shrink. This experiment highlights a possible weak spot of these tumor cells. To understand how FAK makes ovarian cancer cells resistant to chemotherapy, Osterman et al. deleted the gene for FAK from the cells and then looked at how this changed the levels of activation of different genes. They found that, in addition to its effects on cell movement, FAK also activated a group of genes that increase resistance to chemotherapy and repair damaged DNA. This better understanding of how ovarian cancer cells resist chemotherapy could lead to new therapies. In particular, there is now a clinical trial for women with chemo-resistant ovarian cancer in which standard chemotherapy is combined with an inhibitor of the FAK protein.
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- 2019
26. Author response: FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy
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Marjaana Ojalill, Kathleen M. Fisch, Jian Li, Alfredo A. Molinolo, David T. Weaver, Denise C. Connolly, Dwayne G. Stupack, Elizabeth G. Kleinschmidt, Isabelle Tancioni, Duygu Ozmadenci, David D. Schlaepfer, Sean Uryu, Adam Mark, Carlos J. Díaz Osterman, Vihren N. Kolev, Michael T. McHale, L.M. Bean, Guo Fu, Florian J. Sulzmaier, Pekka Rappu, A. Barrie, Christine Jean, Kristin N. Taylor, Balázs Győrffy, Jyrki Heino, Xiao Lei Chen, Edward A. Cordasco, Jonathan A. Pachter, Shulin Jiang, Guorong Xu, and Kristen G. Anderson
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business.industry ,Platinum chemotherapy ,Cancer research ,Medicine ,FAK activity ,business ,Ovarian cancer ,medicine.disease - Published
- 2019
27. H-Ras activation and fibroblast-induced TGF-β signaling promote laminin-332 accumulation and invasion in cutaneous squamous cell carcinoma
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Jyrki Heino, Veli-Matti Kähäri, Pekka Rappu, Elina Siljamäki, Pilvi Riihilä, and Liisa Nissinen
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0301 basic medicine ,Adult ,Male ,Skin Neoplasms ,Adolescent ,Proto-Oncogene Proteins p21(ras) ,03 medical and health sciences ,Mice ,Young Adult ,0302 clinical medicine ,Laminin ,Transforming Growth Factor beta ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Neoplasm Invasiveness ,Fibroblast ,Molecular Biology ,biology ,Chemistry ,Spheroid ,Fibroblasts ,Coculture Techniques ,Up-Regulation ,HaCaT ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer cell ,biology.protein ,Cancer research ,Carcinoma, Squamous Cell ,Immunohistochemistry ,Female ,Cell Adhesion Molecules ,Immunostaining ,Neoplasm Transplantation ,Transforming growth factor ,Signal Transduction - Abstract
Cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer, with increasing incidence worldwide. The molecular basis of cSCC progression to invasive and metastatic disease is still incompletely understood. Here, we show that fibroblasts and transforming growth factor-β (TGF-β) signaling promote laminin-332 synthesis in cancer cells in an activated H-Ras-dependent manner, which in turn promotes cancer cell invasion. Immunohistochemical analysis of sporadic UV-induced invasive human cSCCs (n = 208) revealed prominent cSCC cell specific immunostaining for laminin-332 γ2 chain, located in the majority of cases (90%, n = 173) in the invasive edge of the tumors. To mimic the progression of cSCC we established 3D spheroid cocultures using primary skin fibroblasts and HaCaT/ras-HaCaT human keratinocytes. Our results indicate that in 3D spheroids, unlike in monolayer cultures, TGF-β upregulates laminin-332 production, but only in cells that harbour oncogenic H-Ras. Accumulation of laminin-332 was prevented by both H-Ras knock down and inhibition of TGF-β signaling by SB431542 or RAdKD-ALK5 kinase-defective adenovirus. Furthermore, fibroblasts accelerated the invasion of ras-HaCaT cells through collagen I gels in a Ras/TGF-β signaling dependent manner. In conclusion, we demonstrate the presence of laminin-332 in the invasive front of cSCC tumors and report a new Ras/TGF-β-dependent mechanism that promotes laminin-332 accumulation and cancer cell invasion.
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- 2019
28. PD05-01 EXPLORATION OF THE URINARY PROTEOME IN THE SEARCH FOR NEW BIOMARKERS FOR PROSTATE CANCER: A PROMISING ROLE FOR EXTRACELLULAR VESICLES
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Joeri Tulkens, Glenn Vergauwen, lkka Miinalainen, Finland Jyrki Heino Oulu, Pekka Rappu, Olivier De Wever, Piet Ost, Nicolaas Lumen, Bert Dhondt, Edward Geeurickx, Jan Van Deun, Lien Lippens, and An Hendrix
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Prostate cancer ,business.industry ,Urology ,Urinary system ,Proteome ,medicine ,Cancer research ,Potential source ,Biomarker discovery ,medicine.disease ,business ,Tumor tissue ,Extracellular vesicles - Abstract
INTRODUCTION AND OBJECTIVES:Extracellular vesicles (EV) have raised interest as a potential source of biomarker discovery. We developed techniques for high-purity isolation of tumor tissue EV (tEV)...
- Published
- 2019
29. Epithelial Microvesicles Promote an Inflammatory Phenotype in Fibroblasts
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Gethin Rhys Owen, P. Huang, Liangjia Bi, Ya Shen, Jyrki Heino, Jiarui Bi, Leeni Koivisto, Anne Rokka, Zhejun Wang, Markus Haapasalo, Hannu Larjava, and Lari Häkkinen
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0301 basic medicine ,Cell signaling ,Gingiva ,Enzyme-Linked Immunosorbent Assay ,Biology ,Matrix metalloproteinase ,ta3111 ,Mass Spectrometry ,Extracellular Vesicles ,03 medical and health sciences ,medicine ,Humans ,Secretion ,Interleukin 8 ,Fibroblast ,General Dentistry ,Cells, Cultured ,Periodontal Diseases ,Inflammation ,Regulation of gene expression ,ta313 ,Interleukin-6 ,Interleukin-8 ,Epithelial Cells ,Fibroblasts ,Microvesicles ,Cell biology ,Microscopy, Electron ,Phenotype ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Microscopy, Fluorescence ,Biofilms ,Matrix Metalloproteinase 3 ,Matrix Metalloproteinase 1 ,Signal transduction ,Signal Transduction - Abstract
Microvesicles (MVs) are extracellular vesicles secreted by various cell types that are involved in intercellular communication. We hypothesized that in human periodontal disease, the pocket epithelium releases MVs, which then modulate gene expression in the underlying fibroblasts to control periodontal inflammation. MVs were isolated from culture medium of gingival epithelial cells (GECs) treated with oral bacterial biofilm extract or left untreated. Biofilm treatment significantly increased MV release from the GECs. Mass spectrometry of GEC-MVs identified a total of 2,173 proteins, of which about 80% were detected in MVs from both control and biofilm-treated GECs. Among 80 signature genes of human gingival fibroblasts, 20 were significantly regulated ( P < 0.05) by MVs from control and biofilm-treated GECs in a similar manner. Matrix metalloproteinase 1 and 3 and interleukin 6 and 8 showed the strongest regulation at the mRNA and protein levels. Several cellular signaling pathways were activated by GEC-MVs in human gingival fibroblasts, including Smad and mitogen-activated protein kinase–associated pathways ERK1/2, JNK, and p38. However, ERK1/2 signaling dominated in the MV-induced gene expression changes. The results demonstrate that GEC-MVs have a strong regulatory effect on the expression of fibroblast genes associated with inflammation and matrix degradation and that bacterial biofilm stimulates the generation of GEC-MVs. This suggests that bacterial biofilms can contribute to the initiation and progression of periodontal disease by promoting a tissue-destructive phenotype in gingival fibroblasts via the enhanced secretion of epithelial MVs.
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- 2016
30. Extracellular citrullination inhibits the function of matrix associated TGF-beta
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Hannu Larjava, Mark S. Johnson, Jarmo Käpylä, Laura Pirilä, Pekka Rappu, Vipin Ranga, Kalle Sipilä, Jyrki Heino, and Annamari Torittu
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0301 basic medicine ,Arginine ,Integrin ,Amino Acid Motifs ,CHO Cells ,Biology ,Protein Serine-Threonine Kinases ,ta3111 ,Extracellular matrix ,Arthritis, Rheumatoid ,Transforming Growth Factor beta1 ,03 medical and health sciences ,0302 clinical medicine ,Cricetulus ,Cell Line, Tumor ,Cricetinae ,Extracellular ,Animals ,Humans ,Molecular Biology ,030203 arthritis & rheumatology ,Receptor, Transforming Growth Factor-beta Type II ,ta1182 ,Citrullination ,Cell biology ,Extracellular Matrix ,Fibronectin ,030104 developmental biology ,Ectodomain ,Immunology ,biology.protein ,Protein-Arginine Deiminases ,Protein Processing, Post-Translational ,Receptors, Transforming Growth Factor beta ,Transforming growth factor ,Protein Binding ,Signal Transduction - Abstract
In inflammatory arthritis peptidyl arginine deiminase (PAD) enzymes can citrullinate arginine residues in extracellular matrix (ECM) proteins, such as collagens and fibronectin. This may lead to the generation of anti-citrullinated protein antibodies, important diagnostic markers in rheumatoid arthritis. In addition, the citrullination may directly affect protein function. Based on structural analysis, we found that most ECM-associated growth factors (GFs) have arginine residues in their receptor recognition sites. Thus, they are potential functional targets of extracellular citrullination. To examine this further, we focused on the citrullination of transforming growth factor-βs (TGF-β), well-known ECM-associated GFs. PAD-treatment of CHO-LTBP1 cell derived matrix, rich with TGF-β, decreased the level of TGF-β activity as detected by HaCaT and MLEC-PAI-1/Lu reporter cells. Additional experiments indicated that PAD-treatment inhibits the integrin-mediated TGF-β activation since PAD-treatment decreased the binding of integrin αVβ6 ectodomain as well as integrin-mediated spreading of MG-63 and HaCaT cells to β1-latency associated peptide (TGF-β1 LAP). The citrullination of the RGD site, an important integrin recognition motif, was confirmed by mass spectrometry. Furthermore, the citrullination of active TGF-β1 inhibited its binding to recombinant TGF-β receptor II, and prevented its ability to activate TGF-β signaling. Thus, extracellular PAD activity can affect the function of ECM-associated growth factors by different mechanisms. Importantly, the citrullination of both latent and active TGF-β has the potency to regulate the inflammatory process.
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- 2016
31. Tumor promoter PMA enhances kindlin-2 and decreases vimentin recruitment into cell adhesion sites
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Pekka Rappu, Maria Salmela, Elina Taipalus, Johanna Lilja, Jyrki Heino, Johanna Jokinen, and Henri Niskanen
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0301 basic medicine ,Carcinogenesis ,Integrin ,Vimentin ,macromolecular substances ,Biology ,ta3111 ,Biochemistry ,Cell Line ,Integrin alpha1beta1 ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cricetinae ,Cell Adhesion ,Animals ,Pseudopodia ,Cytoskeleton ,Cell adhesion ,Membrane Proteins ,Cell Biology ,Adhesion ,Neoplasm Proteins ,Cell biology ,Protein Transport ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Phorbol ,biology.protein ,Tetradecanoylphorbol Acetate ,Lamellipodium - Abstract
Phorbol diester PMA (phorbol 12-myristate 13-acetate) is a well-known promoter of tumor progression. PMA also regulates cell adhesion by several mechanisms including conformational activation of integrins and integrin clustering. Here, PMA was shown to induce lamellipodia formation and reorganization of the adhesion sites as well as actin and vimentin filaments independently of integrin preactivation. To further analyze the mechanism of PMA action, the protein composition in the α1β1 integrin/collagen IV adhesion sites was analyzed by mass spectrometry and proteomics. In four independent experiments we observed the reduced recruitment of vimentin in relation to integrin α1 subunit. This was in full agreement with the fact that we also detected the retraction of vimentin from cell adhesions by confocal microscopy. Furthermore, the accumulation of kindlin-2 into cell adhesions was significantly increased after PMA treatment. Kindlin-2 siRNA inhibited cell spreading as well as the formation of actin fibrils and cell adhesions, but did not prevent the effect of PMA on lamellipodia formation. Thus, kindlin-2 recruitment was considered to be a consequence rather than the primary cause for the loss of connection between vimentin and the adhesion sites.
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- 2016
32. Behavior of Alkali-Bearing Minerals in Coking and Blast Furnace Processes
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Stanislav S. Gornostayev, Jyrki Heino, Eetu-Pekka Heikkinen, Timo Fabritius, and Satu Huttunen
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Mineral ,Materials science ,Muscovite ,Metallurgy ,Metals and Alloys ,02 engineering and technology ,Coke ,engineering.material ,Condensed Matter Physics ,020501 mining & metallurgy ,Albite ,chemistry.chemical_compound ,Montmorillonite ,0205 materials engineering ,chemistry ,Illite ,Materials Chemistry ,engineering ,Physical and Theoretical Chemistry ,Leucite ,Biotite - Abstract
The distribution, structural, and textural features, and behavior of alkali-bearing (K and Na) minerals are characterized from coking coals to feed coke and blast furnace (BF) coke. In coals, they are mostly represented by phyllosilicates (illite, montmorillonite, biotite, and muscovite) and feldspars. During the coking process, phyllosilicates retain a layered texture of their aggregates. Under coking conditions, feldspars keep the original outlines of mineral grains and bulk composition. There are no substantial changes in the volume of mineral grains and these alkali-bearing minerals have no physical effect on the coke matrix at coking temperatures. Thermodynamic calculations have shown that there are two major alkali-bearing mineral phases left in the system toward the end of the coking process – albite and leucite. Under BF conditions, Na is fully transferred to the molten phase above 1000 °C, while K partly occurs in leucite up to 1434 °C. The formation of intercalation compounds under BF conditions starts at 1127–1136 °C. Since K is referred to as a main intercalate phase in coke, then attention should primarily be focused on the presence of K-bearing minerals in coking coals. The quality of coals is assessed by the amount of K-bearing phyllosilicates and K-feldspars.
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- 2015
33. Keratinocyte Microvesicles Regulate the Expression of Multiple Genes in Dermal Fibroblasts
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Gethin Owen, Hannu Larjava, Anne Rokka, Jiarui Bi, Lari Häkkinen, Weimin Chen, Jyrki Heino, Leeni Koivisto, and Ping Huang
- Subjects
Keratinocytes ,MAP Kinase Signaling System ,Dermatology ,Biology ,Biochemistry ,Fibroblast migration ,03 medical and health sciences ,0302 clinical medicine ,Transforming Growth Factor beta ,medicine ,Humans ,Extracellular Signal-Regulated MAP Kinases ,Molecular Biology ,Skin ,030304 developmental biology ,Tube formation ,Wound Healing ,0303 health sciences ,integumentary system ,ta1182 ,Granulation tissue ,Transforming growth factor beta ,Cell Biology ,Fibroblasts ,Microvesicles ,Cell biology ,HaCaT ,medicine.anatomical_structure ,Gene Expression Regulation ,030220 oncology & carcinogenesis ,biology.protein ,Keratinocyte ,Wound healing - Abstract
Extracellular vesicles released from cells regulate many normal and pathological conditions. Little is known about the role of epidermal keratinocyte microvesicles (KC-MVs) in epithelial–stromal interaction that is essential for wound healing. We investigated, therefore, whether MV-like structures are present in human wounds and whether they affect wound healing–associated gene expression in dermal fibroblasts. In human wounds, MV-like vesicles were observed during active epithelial migration and early granulation tissue formation. When KC-MVs derived from keratinocyte-like cells (HaCaT) were added to fibroblast cultures, expression of 21 genes was significantly regulated (P
- Published
- 2015
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34. Feasibility of Mechanical Extrusion to Coat Nanoparticles with Extracellular Vesicle Membranes
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Olivier De Wever, Jan Van Deun, Jyrki Heino, An Hendrix, Pekka Rappu, Quentin Roux, Frank Vanhaecke, Thibaut Van Acker, Bruno G. De Geest, Sarah Deville, and Ilkka Miinalainen
- Subjects
0301 basic medicine ,Materials science ,Metal Nanoparticles ,Nanoparticle ,02 engineering and technology ,Article ,Extracellular Vesicles ,Mice ,03 medical and health sciences ,Medicine and Health Sciences ,Animals ,Humans ,cancer ,exosome ,biomimetics ,lcsh:QH301-705.5 ,EXOSOMES ,Microvesicle ,General Medicine ,Extracellular vesicle ,021001 nanoscience & nanotechnology ,Rats ,030104 developmental biology ,Membrane ,lcsh:Biology (General) ,Membrane protein ,Colloidal gold ,GOLD NANOPARTICLES ,drug delivery ,Drug delivery ,Biophysics ,functionalization ,microvesicle ,Surface modification ,DOXORUBICIN DELIVERY ,extracellular vesicle ,0210 nano-technology ,TUMOR MICROENVIRONMENT ,gold nanoparticle - Abstract
Biomimetic functionalization to confer stealth and targeting properties to nanoparticles is a field of intense study. Extracellular vesicles (EV), sub-micron delivery vehicles for intercellular communication, have unique characteristics for drug delivery. We investigated the top-down functionalization of gold nanoparticles with extracellular vesicle membranes, including both lipids and associated membrane proteins, through mechanical extrusion. EV surface-exposed membrane proteins were confirmed to help avoid unwanted elimination by macrophages, while improving autologous uptake. EV membrane morphology, protein composition and orientation were found to be unaffected by mechanical extrusion. We implemented complementary EV characterization methods, including transmission- and immune-electron microscopy, and nanoparticle tracking analysis, to verify membrane coating, size and zeta potential of the EV membrane-cloaked nanoparticles. While successful EV membrane coating of the gold nanoparticles resulted in lower macrophage uptake, low yield was found to be a significant downside of the extrusion approach. Our data incentivize more research to leverage EV membrane biomimicking as a unique drug delivery approach in the near future.
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- 2020
35. Abstract A61: FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy
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Adam A. Mark, Marjaana Ojalill, Guo Fu, Jyrki Heino, Denise C. Connolly, Florian J. Sulzmaier, Guorong Xu, Jonathan A. Pachter, Duygu Ozmadenci, Elizabeth G. Kleinschmidt, A. Barrie, Carlos J. Díaz Osterman, Shulin Jiang, Dwayne G. Stupack, Kristin N. Taylor, Balázs Győrffy, Xiao Lei Chen, David D. Schlaepfer, Alfredo A. Molinolo, Pekka Rappu, David T. Weaver, Michael T. McHale, Kathleen M. Fisch, Jian Li, and L.M. Bean
- Subjects
Cancer Research ,Oncology ,business.industry ,Platinum chemotherapy ,Cancer research ,medicine ,FAK activity ,Ovarian cancer ,medicine.disease ,business - Abstract
Gene copy number alterations, tumor cell stemness, and development of platinum chemotherapy resistance contribute to high-grade serous ovarian cancer (HGSOC) recurrence. Stemness phenotypes involving Wnt-beta-catenin, aldehyde dehydrogenase activities, intrinsic platinum resistance, and tumorsphere formation are here associated with spontaneous genetic gains in KRAS, MYC, and FAK (KMF) genes, in a new aggressive murine model of ovarian cancer. Noncanonical signaling via FAK sustained KMF and human tumorsphere proliferation as well as resistance to cisplatin cytotoxicity. Platinum-resistant tumorspheres can acquire a dependence on FAK for growth. Accordingly, increased FAK tyrosine phosphorylation was observed within HGSOC patient tumors surviving neoadjuvant chemotherapy. Combining a FAK inhibitor with platinum overcame chemoresistance, triggering tumor cell apoptosis. FAK transcriptomic analyses across knockout and reconstituted cells identified 135 genes elevated by a FAK activity-dependent, beta-catenin, and Myc signaling axis including pluripotency and DNA repair genes. Identified target increases in HGSOC tumors may reflect oncogenic FAK signaling. Citation Format: Carlos J. Díaz Osterman, Duygu Ozmadenci, Elizabeth G. Kleinschmidt, Kristin N. Taylor, Allison M. Barrie, Shulin Jiang, Lisa M. Bean, Florian J. Sulzmaier, Jian Li, Xiao Lei Chen, Guo Fu, Marjaana Ojalill, Pekka Rappu, Jyrki Heino, Adam A. Mark, Guorong Xu, Kathleen M. Fisch, David T. Weaver, Jonathan A. Pachter, Balázs Győrffy, Michael T. McHale, Denise C. Connolly, Alfredo Molinolo, Dwayne G. Stupack, David D. Schlaepfer. FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr A61.
- Published
- 2020
36. Enteric Species F Human Adenoviruses use Laminin-Binding Integrins as Co-Receptors for Infection of Ht-29 Cells
- Author
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Niklas Arnberg, Linda Sandblad, B. David Persson, Yoshikazu Takada, Jason Gall, Lynn M. Schnapp, Jyrki Heino, Lars Frängsmyr, A. Paul Mould, Annelie Olofsson, and Anandi Rajan
- Subjects
0301 basic medicine ,Adenoviruses ,media_common.quotation_subject ,viruses ,Integrin ,lcsh:Medicine ,Virus Attachment ,CHO Cells ,Integrin alpha6 ,Article ,Microbiology in the medical area ,Cell Line ,03 medical and health sciences ,Transduction (genetics) ,Cricetulus ,Receptors ,Mikrobiologi inom det medicinska området ,Animals ,Humans ,Internalization ,Receptor ,Laminin binding ,lcsh:Science ,media_common ,RGD motif ,Integrin alpha6beta4 ,Multidisciplinary ,biology ,Chemistry ,Adenoviruses, Human ,lcsh:R ,virus diseases ,Surface Plasmon Resonance ,biology.organism_classification ,eye diseases ,3. Good health ,Cell biology ,Virus ,030104 developmental biology ,Cell culture ,biology.protein ,Receptors, Virus ,lcsh:Q ,Laminin ,HT29 Cells ,Human ,Biotechnology - Abstract
The enteric species F human adenovirus types 40 and 41 (HAdV-40 and -41) are the third most common cause of infantile gastroenteritis in the world. Knowledge about HAdV-40 and -41 cellular infection is assumed to be fundamentally different from that of other HAdVs since HAdV-40 and -41 penton bases lack the αV-integrin-interacting RGD motif. This motif is used by other HAdVs mainly for internalization and endosomal escape. We hypothesised that the penton bases of HAdV-40 and -41 interact with integrins independently of the RGD motif. HAdV-41 transduction of a library of rodent cells expressing specific human integrin subunits pointed to the use of laminin-binding α2-, α3- and α6-containing integrins as well as other integrins as candidate co-receptors. Specific laminins prevented internalisation and infection, and recombinant, soluble HAdV-41 penton base proteins prevented infection of human intestinal HT-29 cells. Surface plasmon resonance analysis demonstrated that HAdV-40 and -41 penton base proteins bind to α6-containing integrins with an affinity similar to that of previously characterised penton base:integrin interactions. With these results, we propose that laminin-binding integrins are co-receptors for HAdV-40 and -41. Originally included in thesis in manuscript form.
- Published
- 2018
37. Proline hydroxylation in collagen supports integrin binding by two distinct mechanisms
- Author
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Pekka Rappu, Johanna Myllyharju, Jarmo Käpylä, Kalle Sipilä, Antti M. Salo, Tiina A. Salminen, Johanna Jokinen, Kati Drushinin, and Jyrki Heino
- Subjects
0301 basic medicine ,collagen ,Proline ,post-translational modification (PTM) ,integrin ,Integrin ,Integrin alpha1 ,Glycobiology and Extracellular Matrices ,Crystallography, X-Ray ,Hydroxylation ,Biochemistry ,Collagen Type I ,Prolyl Hydroxylases ,Collagen receptor ,Extracellular matrix ,03 medical and health sciences ,chemistry.chemical_compound ,Hydroxyproline ,Mice ,0302 clinical medicine ,Cell Adhesion ,Animals ,hydroxyproline ,Molecular Biology ,Integrin binding ,connective tissue ,Binding Sites ,biology ,ta1182 ,cell adhesion ,Cell Biology ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,biology.protein ,Biophysics ,Triple helix ,Protein Binding - Abstract
Collagens are the most abundant extracellular matrix proteins in vertebrates and have a characteristic triple-helix structure. Hydroxylation of proline residues is critical for helix stability, and diminished prolyl hydroxylase activity causes wide-spread defects in connective tissues. Still, the role of proline hydroxylation in the binding of collagen receptors such as integrins is unclear. Here, we isolated skin collagen from genetically modified mice having reduced prolyl 4-hydroxylase activity. At room temperature, the reduced proline hydroxylation did not affect interactions with the recombinant integrin α2I domain, but at 37 °C, collagen hydroxylation correlated with the avidity of α2I domain binding. Of note, LC–MS/MS analysis of isolated skin collagens revealed no major changes in the hydroxyproline content of the main integrin-binding sites. Thus, the disrupted α2I domain binding at physiological temperatures was most likely due to structural destabilization of the collagenous helix. Integrin α2I binding to the triple-helical GFPGER motif was slightly weaker than to GFOGER (O = hydroxyproline). This phenomenon was more prominent when α1 integrin was tested. Integrin α1β1 expressed on CHO cells and recombinant α1I domain showed remarkably slower binding velocity and weaker avidity to GFPGER when compared with GFOGER. Structural modeling revealed the critical interaction between Arg-218 in α1I and the hydroxyproline residue in the integrin-binding motif. The role of Arg-218 was further validated by testing a variant R218D α1I domain in solid-phase binding assays. Thus, our results show that the lack of proline hydroxylation in collagen can affect integrin binding by a direct mechanism and via structural destabilization of the triple helix.
- Published
- 2018
38. Integrin α1 (ITGA1)
- Author
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Jyrki Heino
- Published
- 2018
39. PD05-01 EXPLORATION OF THE URINARY PROTEOME IN THE SEARCH FOR NEW BIOMARKERS FOR PROSTATE CANCER: A PROMISING ROLE FOR EXTRACELLULAR VESICLES
- Author
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Dhondt*, Bert, primary, Vergauwen, Glenn, additional, Van Deun, Jan, additional, Geeurickx, Edward, additional, Lippens, Lien, additional, Tulkens, Joeri, additional, Miinalainen, lkka, additional, Rappu, Pekka, additional, Oulu, Finland, Jyrki Heino, additional, Ost, Piet, additional, De Wever, Olivier, additional, Hendrix, An, additional, and Lumen, Nicolaas, additional
- Published
- 2019
- Full Text
- View/download PDF
40. The composition of prostate core matrisome in vivo and in vitro unveiled by mass spectrometric analysis
- Author
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Jyrki Heino, Pekka Rappu, Elina Siljamäki, Marjaana Ojalill, Pekka Taimen, and Peter J. Boström
- Subjects
0301 basic medicine ,Male ,Proteome ,Urology ,Integrin ,Cell Culture Techniques ,Matrix (biology) ,ta3111 ,Mass Spectrometry ,Cell Line ,Extracellular matrix ,03 medical and health sciences ,0302 clinical medicine ,Collagen VI ,Laminin ,In vivo ,Humans ,Aged ,biology ,Chemistry ,Stem Cells ,Prostate ,Prostatic Neoplasms ,Fibroblasts ,Middle Aged ,In vitro ,Cell biology ,Extracellular Matrix ,030104 developmental biology ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,biology.protein ,Collagen ,Biomarkers - Abstract
Background The composition and organization of extracellular matrix (ECM) are important regulators of cell behavior. In particular in the prostate, this central role of the ECM is further stressed by the fact that several potential markers of prostate stem cells are matrix receptors. Methods We established 12 fibroblastic cell lines from cancerous and non-cancerous areas of six prostates and allowed the cells to produce ECM under cell culture conditions. We also performed a proteome wide analysis of the ECM components by mass spectrometry. To study the in vitro activation of fibroblastic cells we compared the differences between the ECM produced in cell culture by six non-cancerous-tissue-derived fibroblasts and the in vivo matrisome from the corresponding non-cancerous tissue of prostate. Results Our results suggest that at tissue level the ECM is mainly produced by fibroblastic cells and that it contains standard collagen I fibrils and fibril-associated proteins. Beaded-filament forming collagen VI is also abundant and basement membranes potentially contain five laminin subtypes and collagens XV and XVIII. As the main finding, we also detected differences when in vivo and in vitro matrisomes were compared. Only 65 out of 206 proteins were found to be common for both in vivo and in vitro samples. Majority of the 55 proteins, which were solely detected in in vivo samples, were considered to be plasma derived. Eighty-six proteins were solely found from in vitro fibroblast-derived ECM, and most of them were related to matrix remodeling or growth factor action, proposing that the activation of fibroblasts in cell culture may remarkably modify their gene expression profile. Finally, in comparison to traditional 2D in vitro cell culture, the ECM composition of 3D spheroid culture was analyzed. The matrisome in spheroid culture did not resemble the in vivo ECM more closely than in monolayer culture. Conclusions Artificial activation of ECM remodeling seems to be a distinctive feature in in vitro models. In conclusion the constitution of ECM produced by prostate derived fibroblasts in vitro is similar, but not identical to the prostate ECM in vivo as shown here by mass spectrometric analysis.
- Published
- 2017
41. Use of Symbiosis Products from Integrated Pulp and Paper and Carbon Steel Mills: Legal Status and Environmental Burdens
- Author
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Roope Husgafvel, Gary Watkins, H. Nordlund, Jyrki Heino, Olli Dahl, Mikko Mäkelä, and Inga-Liisa Paavola
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Exergy ,Legal status ,Waste management ,020209 energy ,Amendment ,General Social Sciences ,Waste ,02 engineering and technology ,010501 environmental sciences ,Raw material ,01 natural sciences ,Waste treatment ,Greenhouse gas ,0202 electrical engineering, electronic engineering, information engineering ,Environmental science ,media_common.cataloged_instance ,European union ,0105 earth and related environmental sciences ,General Environmental Science ,media_common - Abstract
This study assesses the policy/legal status of both multistream residues and potential secondary products ('symbiosis products') and whether there could be environmental benefits associated with the utilization of residues from integrated pulp and paper and carbon steel mills as raw materials for such secondary products. Waste-related European Union (EU) and Finnish policy and legal instruments were reviewed to identify potential constraints for, and suggested next steps in, the development of potential process industry residue-based symbiosis products. The products were soil amendment pellets, low-grade concrete, and mine filler. A global warming potential (GWP) assessment and an exergy analysis were applied to these potential symbiosis products. Some indicative GWP calculations of greenhouse gas emissions associating similar and/or analogous products based on virgin primary raw materials, more energy-intensive processes, and the alternative treatment of these residues as wastes are also presented. This study addresses GWP, exergy, and legal aspects in a holistic manner to determine the potential environmental benefits of secondary products within the EU legal framework. The GWP assessment and exergy analysis indicate that the utilization of multistream residues causes very low environmental burdens in terms of GWP. The utilization option can have potential environmental benefits in terms of GWP through process replacement and avoided landfilling and waste treatment impacts, as well as potentially through emission reductions from product replacement if suitable and safe applications can be identified. Waste regulation does not define the legal requirements under which utilizing residues in such novel concepts as introduced in this study would be possible, nor how waste status could be removed and product-based legislation be applied to the potential products instead. [ABSTRACT FROM AUTHOR]
- Published
- 2015
42. Fe-Si particles on the surface of blast furnace coke
- Author
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Timo Fabritius, Stanislav S. Gornostayev, Jyrki Heino, and Eetu-Pekka Heikkinen
- Subjects
Blast furnace ,Materials science ,Mechanical Engineering ,Metallurgy ,Metals and Alloys ,Analytical chemistry ,Metallurgical coke ,Coke ,Penetration (firestop) ,Mass ratio ,Surface level ,Tuyere ,Geochemistry and Petrology ,Mechanics of Materials ,Materials Chemistry ,Contact area - Abstract
This study investigates the surface of unpolished samples of blast furnace (BF) coke drilled from the tuyere zone, which hosts Fe−Si particles (mostly Fe3Si) that vary in size, shape, depth of submersion (penetration) into the coke matrix, and contact features with the surface. Based on the shape of the particles and the extent of their contact with the coke matrix, they have been grouped into three major types: (I) sphere-like droplets with limited contact area, (II) semi-spheres with a larger contact area, and (III) irregular segregations with a spherical surface, which exhibit the largest contact area among the three types of particles. Considering the ratio between the height (h) of the particles and half of their length at the surface level (l) along the cross-section, these three types can be characterized as follows: (I) h > l, (II) h ≈ l, and (III) h < l. All the three types of particles can be found near each other. The shape and the extent of the contact depend on the de- gree of penetration of the material into the matrix, which is a function of the composition of the particles. Type (I) particles were initially saturated with Si at an earlier stage and, for that reason, they can react less with carbon in the coke matrix than type (II) and (III), thereby moving faster through the coke cone. Thermodynamic calculations have shown that the temperature interval of 1250-1300°C can be consid- ered the starting point for Si entering into molten iron under quartz-dominated coke ash. Accordingly, the initial pick-up of Si by molten iron can be assumed to be mineral-related. In terms of BF practice, better conditions for sliding Fe−Si droplets through the coke cone are available when they come into contact with free SiO2 concentrated into small grains, and when the SiO2/ΣMexOy mass ratio in the coke ash is high.
- Published
- 2015
43. Cellular recognition and macropinocytosis-like internalization of nanoparticles targeted to integrin α2β1
- Author
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Lotta Bergman, Jyrki Heino, Silja Tiitta, A.-B. Puranen, Pasi Kankaanpää, Mika Lindén, and E. von Haartman
- Subjects
Materials science ,media_common.quotation_subject ,Integrin ,Caveolin 1 ,Nanoparticle ,Endocytosis ,Microscopy, Atomic Force ,Flow cytometry ,law.invention ,Confocal microscopy ,law ,Cell Line, Tumor ,medicine ,Humans ,General Materials Science ,Internalization ,media_common ,Microscopy, Confocal ,medicine.diagnostic_test ,biology ,Pinocytosis ,ta1182 ,Antibodies, Monoclonal ,Silicon Dioxide ,Immunohistochemistry ,Cell biology ,Enterovirus B, Human ,biology.protein ,Nanoparticles ,Integrin alpha2beta1 ,Antibodies, Immobilized ,Intracellular - Abstract
Targeting nanoparticles to desired intracellular compartments is a major challenge. Integrin-type adhesion receptors are connected to different endocytosis routes in a receptor-specific manner. According to our previous observations, the internalization of an α2β1-integrin-echovirus-1 complex takes place via a macropinocytosis-like mechanism, suggesting that the receptor could be used to target nanoparticles to this specific entry route. Here, silica-based nanoparticles, carrying monoclonal antibodies against the α2β1 integrin as address labels, were synthesized. Studies with flow cytometry, atomic force microscopy and confocal microscopy showed the particles to attach to the cell surface via the α2β1 integrin. Furthermore, quantitative analysis of nanoparticle trafficking inside the cell performed with the BioImageXD software indicated that the particles enter cells via a macropinocytosis-like process and end up in caveolin-1 positive structures. Thus, we suggest that different integrins can guide particles to distinct endocytosis routes and, subsequently, also to specific intracellular compartments. In addition, we show that with the BioImageXD software it is possible to conduct sensitive and complex analyses of the behavior of small fluorescent particles inside cells, using basic confocal microscopy images.
- Published
- 2015
44. Platelet response to a small molecule inhibitor of α2β1 integrin is associated with ITGA2 C807T dimorphism
- Author
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Jonna Nieminen, Anne Marjamäki, Liisa Nissinen, Pauli Ollikka, Jyrki Heino, and Pekka Rappu
- Subjects
Adult ,Blood Platelets ,Male ,0301 basic medicine ,Genotype ,Pharmacogenomic Variants ,Integrin ,Plasma protein binding ,Biology ,Polymorphism, Single Nucleotide ,Amino Acid Chloromethyl Ketones ,Collagen receptor ,Mice ,Young Adult ,03 medical and health sciences ,Animals ,Humans ,ta318 ,Platelet ,Allele ,Alleles ,ta1182 ,Hematology ,General Medicine ,Middle Aged ,Molecular biology ,Small molecule ,030104 developmental biology ,biology.protein ,Female ,Collagen ,Integrin alpha2beta1 ,Protein Binding - Abstract
High expression of the collagen receptor, α2β1 integrin, on platelets of ITGA2 807T-allele carriers has been identified as a risk factor for thromboembolic conditions, and α2β1 inhibitors are considered to be potential therapeutic agents. In 59 genotyped individuals, we measured α2 expression levels on platelets and analyzed platelet adhesion to collagen under flow conditions. A sulfonamide-type small-molecule inhibitor of α2β1 integrin decreased average platelet adhesion in individuals with the C/T807T genotype but not in those harboring C807C. Thus, genotype can be used to select a human subpopulation that has the highest probability of showing a positive response to α2β1 inhibitors.
- Published
- 2015
45. Integrin α2β1 in nonactivated conformation can induce focal adhesion kinase signaling
- Author
-
Maria, Salmela, Johanna, Jokinen, Silja, Tiitta, Pekka, Rappu, R Holland, Cheng, and Jyrki, Heino
- Subjects
Protein Conformation ,Science ,Hela Cells ,Focal Adhesion Protein-Tyrosine Kinases ,Medicine ,Humans ,Integrin alpha2beta1 ,Article ,HeLa Cells ,Signal Transduction - Abstract
Conformational activation of integrins is generally required for ligand binding and cellular signalling. However, we have previously reported that the nonactivated conformation of α2β1 integrin can also bind to large ligands, such as human echovirus 1. In this study, we show that the interaction between the nonactivated integrin and a ligand resulted in the activation of focal adhesion kinase (FAK) in a protein kinase C dependent manner. A loss-of-function mutation, α2E336A, in the α2-integrin did not prevent the activation of FAK, nor did EDTA-mediated inactivation of the integrin. Full FAK activation was observed, since phosphorylation was not only confirmed in residue Y397, but also in residues Y576/7. Furthermore, initiation of downstream signaling by paxillin phosphorylation in residue Y118 was evident, even though this activation was transient by nature, probably due to the lack of talin involvement in FAK activation and the absence of vinculin in the adhesion complexes formed by the nonactivated integrins. Altogether these results indicate that the nonactivated integrins can induce cellular signaling, but the outcome of the signaling differs from conventional integrin signaling.
- Published
- 2017
46. Textural changes in metallurgical coke prepared with polyethylene
- Author
-
Timo Fabritius, Satu Huttunen, Stanislav S. Gornostayev, Tommi Kokkonen, Jyrki Heino, and Hannu Makkonen
- Subjects
Materials science ,business.industry ,Mechanical Engineering ,Isotropy ,Metallurgy ,Metals and Alloys ,Coke ,Polyethylene ,Decomposition ,law.invention ,chemistry.chemical_compound ,chemistry ,Optical microscope ,Geochemistry and Petrology ,Mechanics of Materials ,law ,Materials Chemistry ,Coal ,Texture (crystalline) ,High-density polyethylene ,Composite material ,business - Abstract
The effect of high-density polyethylene (HDPE) on the textural features of experimental coke was investigated using polarized-light optical microscopy and wavelet-based image analysis. Metallurgical coke samples were prepared in a laboratory-scale furnace with 2.5%, 5.0%, 7.5%, 10.0%, and 12.5% HDPE by mass, and one sample was prepared by 100% coal. The amounts and distribution of textures (isotropic, mosaic and banded) and pores were obtained. The calculations reveal that the addition of HDPE results in a decrease of mosaic texture and an increase of isotropic texture. Ethylene formed from the decomposition of HDPE is considered as a probable reason for the texture modifications. The approach used in this study can be applied to indirect evaluation for the reactivity and strength of coke.
- Published
- 2014
47. Integrins in periodontal disease
- Author
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Hannu Larjava, Leeni Koivisto, Lari Häkkinen, and Jyrki Heino
- Subjects
Periodontitis ,Integrins ,Cell signaling ,biology ,Integrin ,Junctional epithelium ,Cell Biology ,medicine.disease ,Collagen receptor ,Cell biology ,Mice ,Integrin alpha M ,Immunology ,biology.protein ,medicine ,Animals ,Humans ,Periodontal fiber ,Cell adhesion ,Periodontal Diseases - Abstract
Cell surface integrin receptors mediate cell adhesion, migration and cellular signaling in all nucleated cells. They are activated by binding to extracellular ligands or by intracellular proteins, such as kindlins that engage with their cytoplasmic tails. Cells in the periodontal tissues express several integrins with overlapping ligand-binding capabilities. A distinct phenotype in the periodontium has only been described for knockouts or mutations of three integrin subunits, α11, β6 and β2. Integrin α11β1 appears to have some regulatory function in the periodontal ligament of continuously erupting incisors in mice. Integrin αvβ6 is expressed in the junctional epithelium (JE) of the gingiva. Animals deficient in this receptor develop classical signs of periodontal disease, including inflammation, apical migration of the JE and bone loss, suggesting that it plays a role in the regulation of periodontal inflmmation, likely through activation of transforming growth factor-β1. Lack of integrin activation in the JE is also associated with periodontitis. Patients with kindlin-1 mutations have severe early-onset periodontal disease. Finally, patients with mutations in the leukocyte-specific β2 integrin subunit have severe periodontal problems due to lack of transiting neutrophils in the periodontal tissues.
- Published
- 2014
48. 447 Complement component C1r upregulates the expression of matrix metalloproteinase-1, -13 and -10 and promotes invasion of cutaneous squamous carcinoma cells
- Author
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Marjaana Ojalill, Liisa Nissinen, Seppo Meri, Veli-Matti Kähäri, Kristina Viiklepp, Pilvi Riihilä, and Jyrki Heino
- Subjects
COMPLEMENT COMPONENT C1r ,Chemistry ,Cancer research ,Cell Biology ,Dermatology ,Matrix metalloproteinase ,Molecular Biology ,Biochemistry ,Squamous carcinoma - Published
- 2019
49. Occurrence and Behavior of Phosphorus‐Bearing Minerals in Metallurgical Coke
- Author
-
Stanislav S. Gornostayev, Eetu-Pekka Heikkinen, Jyrki Heino, and Timo Fabritius
- Subjects
Blast furnace ,Bearing (mechanical) ,Materials science ,Phosphorus ,Metallurgy ,Metals and Alloys ,chemistry.chemical_element ,Metallurgical coke ,Coke ,Condensed Matter Physics ,law.invention ,chemistry ,law ,Materials Chemistry ,Physical and Theoretical Chemistry - Published
- 2019
50. Leukocyte integrins αLβ2, αMβ2 and αXβ2 as collagen receptors—Receptor activation and recognition of GFOGER motif
- Author
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Matti Lahti, Jarmo Käpylä, and Jyrki Heino
- Subjects
Receptors, Collagen ,Amino Acid Motifs ,Integrin ,Integrin alphaXbeta2 ,Macrophage-1 Antigen ,HL-60 Cells ,Complement receptor ,Biochemistry ,Collagen receptor ,Extracellular matrix ,Leukemia, Promyelocytic, Acute ,Cell surface receptor ,Cell Line, Tumor ,Cell Adhesion ,Humans ,Binding site ,Antibodies, Blocking ,Cell adhesion ,Binding Sites ,biology ,Cell adhesion molecule ,Chemistry ,Cell Biology ,Molecular biology ,Lymphocyte Function-Associated Antigen-1 ,Protein Structure, Tertiary ,biology.protein ,Collagen ,Protein Binding - Abstract
Integrins α L β 2 , α M β 2 and α X β 2 are expressed on leukocytes. Their primary ligands are counter transmembrane receptors or plasma proteins, such as intercellular cell adhesion molecule-1 (ICAM-1) or components of complement system (iC3b, iC4b), respectively. Function blocking antibodies for these integrins may also reduce cell adhesion to collagens. To make the first systematical comparison of human α L β 2 , α M β 2 and α X β 2 as collagen receptors, we produced the corresponding integrin αI domains both in wild-type and activated form and measured their binding to collagens I–VI. In the “closed” (wild-type) conformation, the α L I and α M I domains bound with low avidity to their primary ligands, and the interaction with collagens was also very weak. Gain-of-function mutations α L I306G, α L K287C/K294C and α M I316G are considered to mimic “open”, activated αI domains. The binding of these activated αI domains to the primary ligands was clearly stronger and they also recognized collagens with moderate avidity ( K d L I domain favored collagen I ( K d ≈ 80 nM) when compared to collagen IV. The integrin α X I domain acted in a very different manner since already in native, wild-type form it bound to collagen IV and iC3b ( K d ≈ 200–400 nM). Antibodies against α X β 2 and α M β 2 blocked promyelocytic leukemia cell adhesion to the collagenous GFOGER motif, a binding site for the β 1 integrin containing collagen receptors. In brief, leukocyte β 2 integrins may act as collagen receptors in a heterodimer specific manner.
- Published
- 2013
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