Differential gene expression during capillary morphogenesis in 3D collagen matrices: regulated expression of genes involved in basement membrane matrix assembly, cell cycle progression, cellular differentiation and G-protein signaling.
We have performed a screening analysis of differential gene expression using a defined in vitro model of human capillary tube formation. Gene array, differential display and cDNA library screening were used to identify both known and novel differentially expressed genes. Major findings include: the upregulation and functional importance of genes associated with basement membrane matrix assembly; the upregulation of growth factors, transcription factors, anti-apoptotic factors, markers of endothelial cell differentiation, JAK-STAT signalling molecules, adhesion receptors, proteinase inhibitors and actin regulatory proteins; and expression changes consistent with inhibition of cell cycle progression, increased cholesterol biosynthesis, decreased ubiquitin-proteasome mediated degradation, and activation of G-protein signaling pathways. Using DNA microarray analysis, the most induced genes at 8, 24 and 48 hours compared with those at 0 hours were jagged-1, stanniocalcin and angiopoietin-2, whereas the most repressed genes were connective tissue growth factor, fibulin-3 and RGS-5. In addition, the full length coding sequence of two novel regulated capillary morphogenesis genes (CMGs) are presented. CMG-1 encodes a predicted intracellular 65 kDa protein with coiled-coil domains. A CMG-1-green fluorescent protein (GFP) chimera was observed to target to an intracellular vesicular compartment. A second novel gene, CMG-2, was found to encode a predicted intracellular protein of 45 kDa containing a transmembrane segment and a CMG-2-GFP chimera was observed to target to the endoplasmic reticulum. A recombinant portion of CMG-2 was found to bind collagen type IV and laminin, suggesting a potential role in basement membrane matrix synthesis and assembly. These data further elucidate the genetic events regulating capillary tube formation in a 3D matrix environment.
Bell
S E
SE
Department of Pathology and Laboratory Medicine, Texas A&M University System Health Science Center, College Station 77843-1114, USA.
Mavila
A
A
Salazar
R
R
Bayless
K J
KJ
Kanagala
S
S
Maxwell
S A
SA
Davis
G E
GE
D000595:Amino Acid Sequence; D001485:Basement Membrane / Q000502:physiology; D002196:Capillaries / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D002784:Cholesterol / Q000096:biosynthesis; D003094:Collagen / Q000494:pharmacology; D024141:Collagen Type IV / Q000096:biosynthesis; D003546:Cysteine Endopeptidases / Q000235:genetics; D003599:Cytoskeleton / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D004730:Endothelium, Vascular / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005346:Fibroblast Growth Factors / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology*; D006133:Growth Substances / Q000235:genetics; D006728:Hormones / Q000235:genetics; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics; D051798:Inhibitor of Differentiation Protein 1; D016023:Integrins / Q000235:genetics; D053613:Janus Kinase 1; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics; D018919:Neovascularization, Physiologic / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010750:Phosphoproteins / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D020710:RGS Proteins / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism; D018000:Receptors, Peptide; D012097:Repressor Proteins; D050794:STAT1 Transcription Factor; D015398:Signal Transduction / Q000502:physiology*; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D051816:Tristetraprolin; D025801:Ubiquitin / Q000235:genetics; D014471:Umbilical Veins / Q000166:cytology
C447150:ANTXR2 protein, human; D024141:Collagen Type IV; D004268:DNA-Binding Proteins; D016326:Extracellular Matrix Proteins; D006133:Growth Substances; D006728:Hormones; C497453:ID1 protein, human; D017874:Immediate-Early Proteins; D051798:Inhibitor of Differentiation Protein 1; D016023:Integrins; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D010750:Phosphoproteins; D020710:RGS Proteins; D018000:Receptors, Peptide; D012097:Repressor Proteins; C486055:SPRY1 protein, human; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D015534:Trans-Activators; D014157:Transcription Factors; D051816:Tristetraprolin; D025801:Ubiquitin; C497552:ZFP36 protein, human; D005346:Fibroblast Growth Factors; D003094:Collagen; D002784:Cholesterol; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; C507904:JAK1 protein, human; D053613:Janus Kinase 1; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D019204:GTP-Binding Proteins