Fibronectin fibrillogenesis regulates three-dimensional neovessel formation

Zhou, Xiaoming; Rowe, R. Grant; Hiraoka, Nobuaki; George, Jerry P.; Wirtz, Denis; Mosher, Deane F.; Virtanen, Ismo; Chernousov, Michael A.; Weiss, Stephen J.

doi:10.1101/gad.1643308

Cited by 186 publications

(172 citation statements)

References 74 publications

Supporting

Mentioning

156

Contrasting

Order By: Relevance

“…4D). Indeed, it is now well established that sustained activa- tion of MAPK signaling pathway is critical for tubulogenesis, whereas transient activation is insufficient to induce such morphological alterations (8,24,25). Phenotypic quantification of the vascular tubes also confirmed that pretreatment of the HUVECs with PHA665752 (10 −7 M), PD98059 (50 μM) or LY294002 (50 μM) blocked 1K1-NP-induced tubulogenesis (Fig.…”

Section: Resultssupporting

confidence: 50%

“…While free 1K1 and 1K1-NP induced similar levels of ERK phosphorylation at early time points, we observed that the nanoparticle resulted in sustained activation of the MAPK signaling in contrast to free 1K1. In a previous study, the sustained activation of MAPK was reported to be critical for tubulogenesis while a transient activation was shown to be sufficient for the induction of cell proliferation (8,(24)(25). This observation can possibly explain that while 1K1 and 1K1-NP exhibited similar levels of cell proliferation, the latter resulted in enhanced neovascularization in the in vivo matrigel implant and zebrafish angiogenesis assay.…”

Section: Resultsmentioning

confidence: 70%

“…A recent review of these clinical trials highlighted vector "washout" leading to inadequate duration of exposure to the angiogenic agent as a key factor that may have impaired the effectiveness of therapy (7). Indeed, it is now established that sustained activation of downstream signal pathways promotes cellular processes critical for angiogenesis (8). We rationalized that harnessing a nanotechnology-based approach can enable a temporal control over the presentation of the angiogenic factors to the cellular target, thereby altering the activation of downstream signal transduction.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Coupling growth-factor engineering with nanotechnology for therapeutic angiogenesis

Roy

Soni

Harfouche

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Therapeutic angiogenesis is an emerging paradigm for the management of ischemic pathologies. Proangiogenic Therapy is limited, however, by the current inability to deliver angiogenic factors in a sustained manner at the site of pathology. In this study, we investigated a unique nonglycosylated active fragment of hepatocyte growth factor/scatter factor, 1K1, which acts as a potent angiogenic agent in vitro and in a zebrafish embryo and a murine matrigel implant model. Furthermore, we demonstrate that nanoformulating 1K1 for sustained release temporally alters downstream signaling through the mitogen activated protein kinase pathway, and amplifies the angiogenic outcome. Merging protein engineering and nanotechnology offers exciting possibilities for the treatment of ischemic disease, and furthermore allows the selective targeting of downstream signaling pathways, which translates into discrete phenotypes.N eovascularization in the adult holds the potential for ameliorating ischemic disease, which is the leading cause of morbidity and mortality in the United States (1). Ferrara and Kerbel recently articulated that "therapeutic angiogenesis" is an emerging and exciting frontier of cardiovascular medicine (2). However, while early preclinical studies and phase I clinical trials exhibited promising results with recombinant proteins or gene therapy, more rigorous phase II and III studies have reported conflicting outcomes (3-6). A recent review of these clinical trials highlighted vector "washout" leading to inadequate duration of exposure to the angiogenic agent as a key factor that may have impaired the effectiveness of therapy (7). Indeed, it is now established that sustained activation of downstream signal pathways promotes cellular processes critical for angiogenesis (8). We rationalized that harnessing a nanotechnology-based approach can enable a temporal control over the presentation of the angiogenic factors to the cellular target, thereby altering the activation of downstream signal transduction.While nanoformulation of angiogenic proteins is an attractive strategy, the synthesis and instability of these large complex glycosylated structures pose a significant barrier (9). Hepatocyte growth factor/scatter factor (HGF/SF) is a potent angiogenic agent (10), signaling through the Met receptor and downstream MAPK and the PI3K pathways (11-12). Interestingly, the Met receptor is upregulated during hypoxia, suggesting that the ischemic tissue will be primed for activation by HGF/SF (13). These results indicate that HGF/SF could emerge as an attractive therapy for angiogenesis. However, HGF/SF is a heparin-binding protein with a complex multidomain structure consisting of an N-terminal (N) domain, four copies of the kringle domain (K1 to K4) and a C-terminal domain homologous to that of serine proteineases (SP) (Fig. 1A) (14). Biologically active HGF/SF is produced by proteolytic cleavage of the linker connecting the fourth kringle and the SP domain yielding a two-chain (α/β)-heterodimer, in which both the α-and the...

show abstract

Section: Resultssupporting

confidence: 50%

Section: Resultsmentioning

confidence: 70%

mentioning

confidence: 99%

See 1 more Smart Citation

Coupling growth-factor engineering with nanotechnology for therapeutic angiogenesis

Roy

Soni

Harfouche

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Normal epidermis, which is avascular and does not depend on FN receptors and ligands, is unaffected by changes in MYO1E or FAK function (6,29). On the other hand, complex mesenchymal processes such as neovascularization require the induction of an FN-based ECM (30) and are affected by FAK (31) or MYO1E loss of function (29). Cancer cells produce an FN-rich fibrotic matrix to facilitate metastasis, and both FAK (4, 7) and MYO1E (32) promote carcinogenesis in murine cancer models.…”

Section: Discussionmentioning

confidence: 99%

Myosin-1E interacts with FAK proline-rich region 1 to induce fibronectin-type matrix

Heim

Squirewell

Neu

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase involved in development and human disease, including cancer. It is currently thought that the four-point one, ezrin, radixin, moesin (FERM)-kinase domain linker, which contains autophosphorylation site tyrosine (Y) 397, is not required for in vivo FAK function until late midgestation. Here, we directly tested this hypothesis by generating mice with FAK Y397-to-phenylalanine (F) mutations in the germline. We found that Y397F embryos exhibited reduced mesodermal fibronectin (FN) and osteopontin expression and died during mesoderm development akin to FAK kinase-dead mice. We identified myosin-1E (MYO1E), an actin-dependent molecular motor, to interact directly with the FAK FERM-kinase linker and induce FAK kinase activity and Y397 phosphorylation. Active FAK in turn accumulated in the nucleus where it led to the expression of osteopontin and other FN-type matrix in both mouse embryonic fibroblasts and human melanoma. Our data support a model in which FAK Y397 autophosphorylation is required for FAK function in vivo and is positively regulated by MYO1E.focal adhesion | myosin | fibronectin | melanoma | cancer F ocal adhesion kinase (FAK) is a nonreceptor tyrosine kinase involved in many biological processes, ranging from mesoderm development to cancer cell metastasis (1). FAK localizes to focal adhesions (2), where it becomes part of a multiprotein complex that links the extracellular matrix (ECM) to the intracellular actin cytoskeleton. FAK is also found in the nucleus, where it is believed to relay information from the cell cortex (3) and induce transcriptional changes (4). The domain architecture of FAK comprises a four-point one, ezrin, radixin, moesin (FERM) domain that is separated from a C-terminal catalytic kinase domain by the FERM-kinase linker. FAK kinase-dead (5) embryos die with mesodermal defects during late gastrulation. In contrast, mice with conditional FAK deletions in the epidermis (6) or breast epithelium (7) show resistance to carcinogenesis.Although FAK has important biological functions, the mechanisms regulating its activity are incompletely understood. For example, it is unclear whether the FERM-kinase linker that contains autophosphorylation site tyrosine (Y) 397 is required for FAK activity in vivo (8). In its closed, inactive conformation, the FAK kinase domain is autoinhibited through interaction with the N-terminal FERM domain. Y397 is nonphosphorylated (9). Upon activation by tethering (10) or other stimuli that induce conformational change (11), the linker region is exposed and Y397 becomes autophosphorylated, leading to the recruitment of the protooncogene SRC. FAK and SRC then form a transient complex, which stabilizes FAK in its active conformation and induces changes in cell shape and focal adhesion turnover in vitro (12). However, mice with a 19-aa deletion in the FAK linker that includes Y397 develop normally until midgestation (8).Here, we have mechanistically discerned the contributions of Y397 to FAK function in viv...

show abstract

“…The established, functional upstream domain (FUD) of Streptococcus pyogenes, which efficiently inhibits fibronectin assembly (Tomasini-Johansson et al, 2001;Ensenberger et al, 2004), and its inactive mutant Del29 lacking amino acid residue I 29 (Zhou et al, 2008), were added at 500 nM to HSFs at the time of seeding (7.5 ϫ 10 4 cells/well) into eight-well chamber slides. After incubation for 48 h, the cells were processed and stained with anti-fibrillin-1 and anti-fibronectin antibodies as described in Immunofluorescence Microscopy.…”

Section: Peptide Inhibition and Protein Binding Monitored By Immunoflmentioning

confidence: 99%

Fibrillin Assembly Requires Fibronectin

Sabatier

Chen²,

Fagotto‐Kaufmann³

et al. 2009

MBoC

Self Cite

225

240

View full text Add to dashboard Cite

Fibrillins constitute the major backbone of multifunctional microfibrils in elastic and nonelastic extracellular matrices. Proper assembly mechanisms are central to the formation and function of these microfibrils, and their properties are often compromised in pathological circumstances such as in Marfan syndrome and in other fibrillinopathies. Here, we have used human dermal fibroblasts to analyze the assembly of fibrillin-1 in dependence of other matrix-forming proteins. siRNA knockdown experiments demonstrated that the assembly of fibrillin-1 is strictly dependent on the presence of extracellular fibronectin fibrils. Immunolabeling performed at the light and electron microscopic level showed colocalization of fibrillin-1 with fibronectin fibrils at the early stages of the assembly process. Protein-binding assays demonstrated interactions of fibronectin with a C-terminal region of fibrillin-1, -2, and -3 and with an N-terminal region of fibrillin-1. The C-terminal half of fibrillin-2 and -3 had propensities to multimerize, as has been previously shown for fibrillin-1. The C-terminal of all three fibrillins interacted strongly with fibronectin as multimers, but not as monomers. Mapping studies revealed that the major binding interaction between fibrillins and fibronectin involves the collagen/ gelatin-binding region between domains FNI 6 and FNI 9 . INTRODUCTIONFibrillins are extracellular matrix components with important functions in elastic and nonelastic tissues including blood vessels, bone, and the eye. Fibrillins, together with the latent transforming growth factor-␤ (TGF-␤)-binding proteins (LTBPs), constitute the fibrillin-LTBP family of proteins (Hubmacher et al., 2006). Fibrillins are ϳ350 kDa in size, are disulfide-rich and glycosylated, and have a characteristic modular structure. The three human fibrillinsfibrillin-1, -2, and -3 -are encoded by different genes and are conserved at the amino acid level both in relation to one another and among species. Fibrillins are mainly composed of tandem arrays of calcium-binding epidermal growth factor-like domains interspersed with TGF-␤-binding protein domains (TB/8-Cys) and hybrid domains Hubmacher et al., 2006). Mutations in fibrillins give rise to the so-called fibrillinopathies, which include Marfan syndrome and autosomal dominant Weill-Marchesani syndrome, both caused by mutations in fibrillin-1, and Beal's syndrome, caused by mutations in fibrillin-2 (Robinson et al., 2006).High-molecular-weight, multiprotein assemblies called microfibrils are the functional units of fibrillins, which serve as a scaffold for the biogenesis of elastic fibers, confer structural integrity to individual organ systems, regulate growth factor signaling of TGF-␤-bone morphogenic protein (BMP) superfamily members and provide limited elasticity to tissues (Kielty et al., 2002;Charbonneau et al., 2004;Ramirez and Dietz, 2007). Extracted microfibrils from cell culture or tissues display a typical bead-on-a-string ultrastructure, having a 50 -55-nm periodicity when analyzed by ele...

show abstract

Fibronectin fibrillogenesis regulates three-dimensional neovessel formation

Cited by 186 publications

References 74 publications

Coupling growth-factor engineering with nanotechnology for therapeutic angiogenesis

Coupling growth-factor engineering with nanotechnology for therapeutic angiogenesis

Myosin-1E interacts with FAK proline-rich region 1 to induce fibronectin-type matrix

Fibrillin Assembly Requires Fibronectin

Contact Info

Product

Resources

About