Jean Claude Monboisse

The Antitumor Properties of the α3(IV)-(185-203) Peptide from the NC1 Domain of Type IV Collagen (Tumstatin) Are Conformation-dependent

Tumor progression may be controlled by various fragments derived from noncollagenous 1 (NC1) C-terminal domains of type IV collagen. We demonstrated previously that a peptide sequence from the NC1 domain of the α3(IV) collagen chain inhibits the in vitro expression of matrix metalloproteinases in human melanoma cells through RGD-independent binding to αvβ3 integrin. In the present paper, we demonstrate that in a mouse melanoma model, the NC1 α3(IV)-(185-203) peptide inhibits in vivo tumor growth in a conformation-dependent manner. The decrease of tumor growth is the result of an inhibition of cell proliferation and a decrease of cell invasive properties by down-regulation of proteolytic cascades, mainly matrix metalloproteinases and the plasminogen activation system. A shorter peptide comprising the seven N-terminal residues 185-191 (CNYYSNS) shares the same inhibitory profile. The three-dimensional structures of the CNYYSNS and NC1 α3(IV)-(185-203) peptides show a β-turn at the YSNS (188-191) sequence level, which is crucial for biological activity. As well, the homologous MNYYSNS heptapeptide keeps the β-turn and the inhibitory activity. In contrast, the DNYYSNS heptapeptide, which does not form the β-turn at the YSNS level, is devoid of inhibitory activity. Structural studies indicate a strong structure-function relationship of the peptides and point to the YSNS turn as necessary for biological activity. These peptides could act as potent and specific antitumor antagonists of αvβ3 integrin in melanoma progression.

Matrikines from basement membrane collagens: a new anti-cancer strategy.

BACKGROUNDnTumor microenvironment is a complex system composed of a largely altered extracellular matrix with different cell types that determine angiogenic responses and tumor progression. Upon the influence of hypoxia, tumor cells secrete cytokines that activate stromal cells to produce proteases and angiogenic factors. In addition to stromal ECM breakdown, proteases exert various pro- or anti-tumorigenic functions and participate in the release of various ECM fragments, named matrikines or matricryptins, capable to act as endogenous angiogenesis inhibitors and to limit tumor progression.nnnSCOPE OF REVIEWnWe will focus on the matrikines derived from the NC1 domains of the different constitutive chains of basement membrane-associated collagens and mainly collagen IV.nnnMAJOR CONCLUSIONSnThe putative targets of the matrikine control are the proliferation and invasive properties of tumor or inflammatory cells, and the angiogenic and lymphangiogenic responses. Collagen-derived matrikines such as canstatin, tumstatin or tetrastatin for example, decrease tumor growth in various cancer models. Their anti-cancer activities comprise anti-proliferative effects on tumor or endothelial cells by induction of apoptosis or cell cycle blockade and the induction of a loss of their migratory phenotype. They were used in various preclinical therapeutic strategies: i) induction of their overexpression by cancer cells or by the host cells, ii) use of recombinant proteins or synthetic peptides or structural analogues designed from the structure of the active sequences, iii) used in combined therapies with conventional chemotherapy or radiotherapy.nnnGENERAL SIGNIFICANCEnCollagen-derived matrikines strongly inhibited tumor growth in many preclinical cancer models in mouse. They constitute a new family of anti-cancer agents able to limit cancer progression. This article is part of a Special Issue entitled Matrix-mediated cell behaviour and properties.

Transforming growth factor-β1 inhibits tumor growth in a mouse melanoma model by down-regulating the plasminogen activation system

The degradation of basement membranes by tumor cells involves secretion and activation of proteinases, such as matrix metalloproteinases (MMPs) and the plasminogen activation system (uPA, tPA, PAI-1), and results from an imbalance between their inhibitors and activators, controlled by various growth factors or cytokines. Among them, the TGF-beta family is one of the most intriguing because it has been reported either to decrease or promote cancer progression. In the present paper, we studied the effect of TGF-beta1 in a mouse melanoma model. In vivo, TGF-beta1 inhibited tumor growth after subcutaneous injection of B16F1 cells in syngenic mice. In vitro, TGF-beta1 did not alter B16F1 cell proliferation, but strongly decreased their migration through Matrigel-coated membranes. The protease production was analyzed by zymography, Western blot, or RT-PCR. MMP-2 and TIMP-2 expression were not altered by TGF-beta1. In contrast, TGF-beta1 triggered a large decrease of uPA and tPA, as well as a decrease of uPA and uPAR mRNAs. By Western blot and RT-PCR analyses, TGF-beta1 was shown to induce a strong increase of PAI-1 synthesis. Collectively, these results suggest that TGF-beta1 may inhibit melanoma tumor growth by specifically decreasing plasmin activity of tumor cells and play a protective role during the earliest stages of tumor progression.

Malondialdehyde binding to proteins dramatically alters fibroblast functions

The regulation of cell metabolism by the surrounding environment is deeply altered by the posttranslational nonenzymatic modifications of extracellular proteins that occur throughout lifespan in vivo and modify their structural and functional properties. Among them are protein adducts formed by components generated from oxidative processes, such as malondialdehyde (MDA). We have investigated here the effects of MDA‐binding to proteins on cultured fibroblast functions. Type I collagen and/or serum proteins were incubated with 0–100 mM MDA for 3 h before use in fibroblast cultures. In tridimensional lattice cultures, MDA‐treated collagen inhibited the contracting activity of fibroblasts. A similar inhibition of lattice contraction was reproduced by the addition of MDA‐treated serum to the culture medium. In monolayer cultures, the addition of MDA‐modified serum proteins completely inhibited fibroblast multiplication without effect on initial adhesion steps. MDA‐modified proteins decreased the proliferative capacities of cells, strongly altered cell cycle progression by blocking passage to G2/M phases, and induced apoptotic features in fibroblasts. Our results show, for the first time, that MDA‐modified proteins are potentially as deleterious as free MDA, and could be involved in aging as well as in degenerative complications of diseases with increased oxidative stress such as diabetes mellitus or atherosclerosis. J. Cell. Physiol. 191: 227–236, 2002.

Tetrastatin, the NC1 domain of the α4(IV) collagen chain: a novel potent anti-tumor matrikine.

Background NC1 domains from α1, α2, α3 and α6(IV) collagen chains were shown to exert anti-tumor or anti-angiogenic activities, whereas the NC1 domain of the α4(IV) chain did not show such activities so far. Methodology/Principal Findings We demonstrate in the present paper that the NC1 α4(IV) domain exerts a potent anti-tumor activity both in vitro and in an experimental human melanoma model in vivo. The overexpression of NC1 α4(IV) in human UACC-903 melanoma cells strongly inhibited their in vitro proliferative (–38%) and invasive (–52%) properties. MT1-MMP activation was largely decreased and its cellular distribution was modified, resulting in a loss of expression at the migration front associated with a loss of migratory phenotype. In an in vivo xenograft model in athymic nude mice, the subcutaneous injection of NC1 α4(IV)-overexpressing melanoma cells induced significantly smaller tumors (–80% tumor volume) than the Mock cells, due to a strong inhibition of tumor growth. Exogenously added recombinant human NC1 α4(IV) reproduced the inhibitory effects of NC1 α4(IV) overexpression in UACC-903 cells but not in dermal fibroblasts. An anti-αvβ3 integrin blocking antibody inhibited cell adhesion on recombinant human NC1 α4(IV) substratum. The involvement of αvβ3 integrin in mediating NC1 α4(IV) effect was confirmed by surface plasmon resonance (SPR) binding assays showing that recombinant human NC1 α4(IV) binds to αvβ3 integrin (KDu200a=u200a148±9.54 nM). Conclusion/Significance Collectively, our results demonstrate that the NC1 α4(IV) domain, named tetrastatin, is a new endogenous anti-tumor matrikine.

Analytical methods for measuring collagen XIX in human cell cultures, tissue extracts, and biological fluids

Type XIX collagen is a minor collagen associated with basement membranes in vascular, neuronal, mesenchymal, and epithelial tissues. We demonstrated that the NC1, C-terminal, domain of collagen XIX inhibits the migration capacities of tumor cells and exerts a strong inhibition of tumor growth. Other basement membrane collagens or derived fragments were measured in biological fluids such as blood and urine of patients and appeared to be useful for diagnosis, prognosis, or treatment monitoring. The aim of this study was to develop and validate methods to measure collagen XIX and its fragments in human cell cultures, tissue extracts, and human biological fluids. For that purpose, we developed real-time PCR, Western blot, and competitive enzyme-linked immunosorbent assays. We demonstrated that the methods developed in this paper are specific for collagen XIX. We showed that it is expressed in human cell cultures, tissue extracts, and various biological fluids. These methods may be used in various human tissue extracts and biological fluids such as serum, amniotic fluid, cord blood, and many other fluids. Collagen XIX or its fragments could constitute new biomarkers for human diseases as well as for diagnosis and/or prognosis.

Evaluation of Lumipulse® G1200 for the measurement of six tumor markers: Comparison with AIA® 2000

Tumor marker assays are daily practiced, for screening and follow up of cancers. Interassay precision is an important parameter for the interpretation of the kinetics of the markers, in order to conclude to the efficiency or failure of treatment. The aim of this study was to compare two automated Immunoassay analyzers, Lumipulse® G1200 and AIA® 2000. Both analyzers used an immunoassay system but with different antibodies. Six tumor markers commonly used were studied: AFP, PSA, CA 19-9, CA 15-3, CA 125 and CEA. 253 samples have been collected over a period of one month and analyzed by both analyzers. Regression of Passing-Badblock and Bland-Altman diagram were used to analyze the results for AFP (n=36), PSA (n=39), CA-125 (n=40), CA 15-3 (n=40), CA 19-9 (n=46) and CEA (n=52) were performed. Analytical performances of Lumipulse® G1200 highlighted the good inter-run and intra-run precision of the analyzer. We obtained a good correlation coefficient between Lumipulse G1200® and AIA 2000®, >0.96 for most markers except CA 19-9 which provided a correlation coefficient significantly lower than that obtained with other markers. The concordance for all markers was >94% except for CA 19-9 (83.7%). This study showed a good correlation between the two analyzers and, therefore, a transfer from one analyzer to the other is possible for the different markers studied. However, we found here the classical difficulty to transfer this type of analysis, due to the absence of method standardization. This difficulty was particularly illustrated by CA19-9.

Control of Tumor Progression by Extracellular Matrix Molecule Fragments, the Matrikines

Tumor microenvironment is a complex system composed of a largely altered Extracellular Matrix (ECM) with different cell types that determine tumor progression. Upon the influence of hypoxia, tumor cells secrete cytokines that activate stromal cells to produce proteases and angiogenic factors. The proteases degrade the stromal ECM and participate in the release of various ECM fragments, named matrikines or matricryptins, capable to control tumor invasion and metastasis dissemination. The putative targets of the matrikine action are the proliferation and invasive properties of tumor or inflammatory cells, and the angiogenic and lymphangiogenic responses. In the present review, we will describe pro-tumorigenic effects triggered by soluble elastin or Elastin-Derived Peptides (EDPs), as well as the anti-tumorigenic or anti-angiogenic activities the matrikines derived from basement membrane associated collagens and several proteoglycans such as perlecan or lumican. Matrikines constitute a new family of potent anticancer agents that could be used under various therapeutic strategies: i) induction of their overexpression by cancer cells or by the host, ii) use of recombinant proteins or synthetic peptides or structural analogs designed from the structure of the active sequences. Matrikines could be used in combination with conventional chemotherapy or radiotherapy to limit tumor progression.

The Interaction of Human Neutrophils with Type IV Collagen Involves an Inhibitory Signal Transduction Pathway

Type IV collagen is a heterotrimer composed of 3 a chains; these molecules form tetramers by overlap at the amino terminus (7S domain) which further aggregate by end-to-end interaction involving the non-collagenous carboxyl termini (NC1 domains). Collagen is a major component of basement membranes. There are six a chains of type IV collagen whose genes have been cloned. The most predominent heterotrimer is composed of two a-1 and one a-2 chains. It can be prepared by extraction of a mouse sarcoma (EHS tumor) (Wisdom, Gunwar, Hudson, Noelken and Hudson, 1992). Molecular forms of the other type IV collagen chains, a3(IV), a4(IV), a5(IV), a6(IV), have not been isolated (Hudson, Reeders and Tryggwason, 1993; Zhou, Ding, Zhao and Reeders, 1994) and the clones for the a3(IV), a4(IV), a5(IV) and a6(IV) have been reported (Zhou et al., 1994). There is evidence that the latter are also distributed in several types of basement membranes including the kidney glomerulus, the alveolus, the choroid plexus and lens capsule (Ninomiya, Kagawa, Iyama, Naito, Kishiro, Seyer, Sugimoto, Oohashi and Sado, 1995). Lens capsule collagen may be obtained in purified forms. It contains all the types of chains except a6(IV).