Research - Institute of Biochemistry - Eukaryotic Molecular Biology Unit - Laboratory of Extracellular Matrix Biology

Ibolya KISS
Head, Principal Investigator

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Ferenc DEÁK Principal Investigator
Viktória SZŰTS Staff Scientist
Tibor SZÉNÁSI PhD Student
Ferencné SIMON Technician

MOLECULAR BIOLOGY OF THE EXTRACELLULAR MATRIX

Multicellular organisms deposit a highly organized extracellular matrix (ECM) around the cells, which provides physical support and the necessary milieu for normal cell metabolism and development, and delineates pathways during differentiation and tissue regeneration. The ECM performs essential, but very divergent functions in the various tissues and organs. We investigate how matrilins and other noncollagenous glycoproteins involved in the organization of the ECM contribute to tissue integrity, differentiation and carcinogenesis using molecular biology methods, cell culture and transgenesis. We also investigate how the tissue-specific expression of these genes is regulated in the developing musculo-skeletal system.


The function of ECM proteins in muscle development, muscle regeneration, healthy and diseased heart, and carcinogenesis

By cloning the matrilin-1 and matrilin-2 genes, we discovered the matrilin family of noncollagenous proteins, which form collagen-dependent and independent filamentous networks in the ECM of various tissues. Matrilin-1 and -3 are deposited only in cartilage and bone, whereas matrilin-2 and -4 are found in a large variety of tissues including heart, skin, several epithelia, muscle and the nerve system. We study the expression and function of matrilins during differentiation and regeneration processes and under pathological conditions. To this end, we generated cell lines that produce a variety of matrilin-2 recombinants to pinpoint the role of the various protein modules in macromolecular interactions during the organization of the ECM. In the framework of international collaborations, transgenic mice have been generated that are deficient of the matrilin-2 gene. At present our efforts are focused on two projects based on national and international collaborations. The first project aims to study the role of matrilin-2 in muscle differentiation and regeneration. We observed transient activation of the gene in regenerating rat skeletal muscle. In vitro differentiation of myoblasts recapitulated the process. We perform comprehensive analysis to compare matrilin-2 gene expression with those of other marker genes during muscle development and skeletal muscle regeneration in wild type and matrilin-2 deficient mice as well as in differentiating myoblast cultures. It is under study whether matrilin-2 can directly or indirectly affect muscle cells, by interacting with other ECM components. We are interested in the signal transduction pathways affected by matrilin-2. In the second project, we investigate the role of ECM proteins including matrilin-2 in heart development and in dilated cardiomyopathy. We also examine the expression, distribution and interaction of ion channels (e.g. Kir2.x) in healthy and diseased heart. The third project aims to study the role of matrilin-2 in tumor formation. We have found previously that the matrilin-2 protein/RNA level can be used as a diagnostic/prognostic marker in particular neoplasias. Therefore we investigate tumor formation and the underlying molecular mechanisms in matrilin-2 deficient mice. We also participate in testing anticancer drugs.


Molecular basis of cartilage-specific gene expression and establishment of transgenic animal models for studying skeletal development and cartilage regeneration

The prevalence of joint diseases increases worldwide with the aging of the population. Inherited joint diseases disturbing longitudinal bone growth often cause dwarfism, which cannot be cured by current medical treatments. Arthritis is a chronic degenerative arthropathy that frequently leads to persistent pain and disability. Due to the large burden on the society, finding better treatments for arthritis is a major focus of medically oriented research. There is a great need for new therapies that address not only the reduction of pain and inflammation, but also the promotion of cartilage matrix regeneration. Development of new drugs requires the development of animal models suitable for testing cartilage-specific gene expression and cartilage regeneration and protocols need to be tested in. Therefore, we work on establishing transgenic animals for monitoring cartilage wear and regeneration.

Previously we cloned genes for cartilage link protein and matrilin-1 specifically expressed in cartilage. The matrilin-1 gene has the unique property among cartilage protein genes that its expression is restricted to the late proliferative and prehypertrophic zones of the growth plate, which play an important role in the regulation of bone growth. By means of recombinant DNA technology, in vitro DNA-protein interaction studies, transiently transfected cell cultures and transgenic mice, we characterized the regulatory regions and transcription factors responsible for the tissue- and zone-specific expression of the gene. Our data demonstrate a complex regulation based on the modular arrangement of the tissue-specific control elements. We constructed cartilage-specific vectors and generated transgenic mice, which express reporter gene in distinct zones of the growth plate. These animal models are suitable for testing the effect of drugs and drug candidates on skeletal development in safety pharmacology.

We also aim at inducing arthritis in transgenic mice expressing the reporter gene in each chondrocyte and developing an animal model suitable for testing the effect of anti-inflammatory drugs and potential antirheumaticums on articular cartilage degradation and regeneration.

Selected publications

Kiss, I., Deák, F., Mestric, S., Delius, H., Soós, J., Dékány, K., Argraves, W.S., Sparks, K.J. and Goetinck, P.F. (1987). Structure of the chicken link protein gene: Exons correlate with the protein domains. Proc. Natl. Acad. Sci. U.S.A. 84: 6399 6403.

Kiss, I., Deák, F., Holloway, R.G., Jr., Delius, H., Mebust, K.A., Frimberger, E., Argraves, W.S., Tsonis, P.A., Winterbottom, N. and Goetinck, P.F. (1989). Structure of the gene for cartilage matrix protein, a modular protein of the extracellular matrix. J. Biol. Chem. 264: 8126 8134.

Szabó, P., Moitra, J., Rencendorj, A., Rákhely, G., Rauch, T. and Kiss, I. (1995). Identification of a nuclear factor-I family protein-binding site in the silencer region of the cartilage matrix protein gene. J. Biol. Chem. 270: 10212-10221.

Deák, F., Piecha, D., Bachrati, C., Paulsson, M. and Kiss, I. (1997). Primary structure and expression of matrilin-2, the closest relative of cartilage matrix protein within the von Willebrand factor type A-like module superfamily. J. Biol. Chem. 272: 9268-9274.

Deák, F., Wagener, R., Kiss, I. and Paulsson, M. (1999). The matrilins: a novel family of oligomeric extracellular proteins. Matrix Biol. 18: 55-64.

Piecha, D., Muratoglu, S., Mörgelin, M., Hauser, N., Studer, D., Kiss, I., Paulsson, M., Deák, F. (1999). Matrilin-2, a large oligomeric matrix protein, is expressed by a great variety of cells and forms fibrillar networks. J. Biol. Chem. 274: 13353-13361.

Mátés, L., Deák, F., Korpos, P.G.É., Liu, Z., Beier, D.R., Aszódi, A. and Kiss, I. (2002). Comparative analysis of the mouse and human genes (Matn2 and NATN2) for matrilin-2, a filament-forming protein widely distributed in extracellular matrices. Matrix Biol. 21: 163-174.

Karcagi, I., Rauch, T., Hiripi, L., Rentsendorj, O., Nagy, A., Bősze, Zs. and Kiss, I. (2004). Functional analysis of the regulatory regions of the matrilin-1 gene in transgenic mice reveals modular arrangement of tissue-specific control elements. Matrix Biol. 22: 605-618.

Mátés, L., Mörgelin, M., Deák, F., Kiss, I. and Aszódi, A. (2004). Mice lacking the extracellular matrix adaptor protein matrilin-2 develop without obvious abnormalities. Matrix Biol. 23: 195-204.

Korpos, É., Molnár, A., Papp, P., Kiss, I., Orosz, L. and Deák, F. (2005). Expression pattern of matrilins and other extracellular matrix proteins characterize distinct stages of cell differentiation during antler development. Matrix Biol. 24: 124-135.

Rentsendorj, O., Nagy, A., Sinkó, I., Daraba, A., Barta, E. and Kiss, I. (2005). Highly conserved proximal promoter element harbouring paired Sox9-binding sites contributes to the tissue- and developmental stage-specific activity of the matrilin-1 gene. Biochem. J. 389: 705-716.

Sharma, M.K., Watson, M.A., Lyman, M., Perry, A., Aldape, K.D., Deák, F. and Gutmann, D.H. (2006). Matrilin-2 expression distinguishes clinically relevant subsets of pylocytic astrocytoma. Neurology 66: 127-130.

Szabó, E., Lódi, C., Korpos, É., Batmunkh, E., Rottenberger, Z., Deák, F., Kiss, I., Tőkés, A., Lotz, G., László, V., Kiss, A., Schaff, Z. and Nagy, P. (2007). Expression of matrilin-2 in oval cells during rat liver regeneration. Matrix Biol. 26: 554-560.

Molnár, A., Gyurján, I., Korpos, É., Borsy, A., Stéger, V., Buzás, Z., Kiss, I., Zomborszky, Z., Papp, P., Deák, F., Orosz, L. (2007) Identification of differentially expressed genes in the developing antler of red deer Cervus elaphus. Mol. Genet. Genomics 277, 237-248

Keller-Pinter A, Bottka S, Timar J, Kulka J, Katona R, Dux L, Deak F, Szilak L. (2010) Syndecan-4 promotes cytokinesis in a phosphorylation-dependent manner. Cell Mol Life Sci. In press, Mar 14. [Epub ahead of print] PMID: 20229236

S. Bendahhou, C. Marionneau, K. Haurogne, M-M Laroque, R. Derand, V. Szűts, D. Escande, Sophie Demolombe. In vitro molecular interactions and distribution of KCNE family with KCNQ1 in the human heart. Cardiovascular Research, 2005, 67, 529-538.

Molnár A., Tóth A., Bagi Z., Papp Z., Edes I., Vaszily M., Galajda Z., Papp J.G., Varró A., Szűts V., Lacza Z., Gero D., Szabo C. (2006) Activation of the poly (ADP-ribose) polymerase pathway in human heart failure. Mol. Med., 12,143-152.

Nathalie Gaborit, Sabrina Le Bouter, Viktoria Szűts, Andras Varro, Stanley Nattel, Denis Escande, Sophie Demolombe (2007) Regional and Tissue Specific Transcript Signatures of Ion Channel Genes in the Normal Human Heart. J. Physiol., 582, 675-693.

Nagy, N., Szuts, V., Horvath, Z., Seprenyi, G., Farkas, AS., Acsai, K., Prorok, J, Bitay, M,Kun, A., Pataricza, J.,Papp, JG.,Nanasi, PP., Varro, A. Toth, A. (2009) Does small-conductance calcium-activated potassium channel contribute to cardiac repolarization? J. Mol. and Cell. Card., 47, 656-663.

Gaborit, N , Wichter, T , Varro, A, Szuts, V. Lamirault, G., Eckardt, L., Paul, M., Breithardt, G.m, Schulze-Bahr, E ., Escande, D., Nattel, S., Demolombe, S (2009) Transcriptional profiling of ion channel genes in Brugada syndrome and other right ventricular arrhythmogenic diseases. Eur. Heart J., 30, 487-496