Trends in Molecular Medicine
Towards high-throughput functional target discovery in angiogenesis research
Section snippets
Identification of targets for angiogenesis modulation
Angiogenesis, the formation of novel blood vessels from pre-existing blood vessels, is a hallmark of numerous pathologies such as cancer, rheumatoid arthritis, endometriosis, atherosclerosis and diabetic retinopathy 1, 2. In normal tissues, blood vessels and capillaries are generally in a resting state, which is maintained by a fine-tuned balance of pro-angiogenic and anti-angiogenic factors present in the microenvironment. However, different stimuli can disturb this balance in favor of
Target validation: from gene to function
Gene- and protein-expression-profiling techniques have been applied to study molecular events in angiogenesis [6]. Differentially expressed molecules might serve as targets for therapeutic intervention. Validating the relevance of a target for therapeutic applications requires the establishment of a causative relationship between protein expression and/or function and the disease phenotype. This can be achieved by either blocking protein function or modulating protein-expression levels. In this
Functional-target discovery: from function to gene
The wealth of gene-expression data generated to date has presented the scientific community with the formidable challenge of functionally annotating the individual genes in a multitude of processes and diseases. Recently, diverse high-throughput methods have been shown to functionally annotate putative target molecules, overcoming these limitations. Advances in gene-library generation, transfection methods, miniaturization of assays, and screening techniques have provided the opportunity for
Functional genomics in angiogenesis research
Recent advances in high-throughput functional genomics will have a major impact on our future understanding of pathological and physiological angiogenesis. The shift in focus towards functional identification of putative therapeutic targets enables more-efficient clinical translation. Most techniques described in this review rely on efficient transfection of cells. This can be a complicating factor when studying some cell types, such as ECs. Recent developments in high-throughput
Concluding remarks
Functional validation tools, originally developed for the analysis of individual genes, have become useful for high-throughput functional identification with almost no limitations. In conclusion, the use of high-throughput gene-expression analysis, which only generates associative data rather than causative data, is being caught up by novel methods that enable direct functional linkage between the gene-expression level and the phenotype. Although technical challenges remain, recent progress has
Acknowledgements
This work was supported by a grant from the Research Institute for Growth and Development (GROW), Maastricht University. The study has been financially supported by the sixth EU Framework Programme (Integrated Project ‘Angiotargeting’; contract no. 504743) in the area of ‘Life sciences, genomics and biotechnology for health’.
Glossary
- Antisense oligodeoxynucleotide (ODN):
- a single-stranded, DNA-like molecule of 17–22 nucleotides in length that is complementary to its target mRNA and blocks protein translation by forming DNA–RNA duplexes.
- Array:
- a systematically arranged repertoire of DNA, cells or tissues. These arrays can be arranged in microtiter plates, or printed on glass slides or chips (microarray).
- cDNA library:
- a collection of cDNA sequences in separate vectors.
- Expression library:
- a collection of cDNA sequences
References (66)
Vascular gene expression in nonneoplastic and malignant brain
Am. J. Pathol.
(2004)- et al.
In silico analysis of angiogenesis associated gene expression identifies angiogenic stage related profiles
Biochim. Biophys. Acta
(2005) Modulation of angiogenesis with siRNA inhibitors for novel therapeutics
Trends Mol. Med.
(2005)- et al.
Effective intracellular delivery of oligonucleotides in order to make sense of antisense
J. Control. Release
(2004) VEGF antisense therapy inhibits tumor growth and improves survival in experimental pancreatic cancer
Surgery
(2005)Analysis of HeyL expression in wild-type and Notch pathway mutant mouse embryos
Mech. Dev.
(2000)Transfection microarray of human mesenchymal stem cells and on-chip siRNA gene knockdown
J. Control. Release
(2004)Stable suppression of tumorigenicity by virus-mediated RNA interference
Cancer Cell
(2002)A quantitative high-throughput endothelial cell migration assay
J. Biomol. Screen.
(2004)A review of the issues in the pharmacokinetics and toxicology of phosphorothioate antisense oligonucleotides
Biochim. Biophys. Acta
(1999)
Antisense oligonucleotides with different backbones. Modification of splicing pathways and efficacy of uptake
J. Biol. Chem.
Angiogenesis: potentials for pharmacologic intervention in the treatment of cancer, cardiovascular diseases, and chronic inflammation
Pharmacol. Rev.
Tumor angiogenesis
Alterations in vascular gene expression in invasive breast carcinoma
Cancer Res.
Genes expressed in human tumor endothelium
Science
Angiogenesis assays: a critical overview
Clin. Chem.
Quantitative angiogenesis assays in vivo – a review
Angiogenesis
Quantitative monitoring of gene expression patterns with a complementary DNA microarray
Science
The sequence of the human genome
Science
Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells
Nature
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Nature
RNA silencing: the genome's immune system
Science
A system for stable expression of short interfering RNAs in mammalian cells
Science
U6 promoter-driven siRNAs with four uridine 3′ overhangs efficiently suppress targeted gene expression in mammalian cells
Nat. Biotechnol.
Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells
Genes Dev.
Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells
Nat. Biotechnol.
Small interfering RNA production by enzymatic engineering of DNA (SPEED)
Proc. Natl. Acad. Sci. U. S. A.
Restriction enzyme-generated siRNA (REGS) vectors and libraries
Nat. Genet.
Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells
Proc. Natl. Acad. Sci. U. S. A.
An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division
Nature
Down-regulation of vascular endothelial cell growth factor-C expression using small interfering RNA vectors in mammary tumors inhibits tumor lymphangiogenesis and spontaneous metastasis and enhances survival
Cancer Res.
Gene expression profile in fibroblast growth factor 2-transformed endothelial cells
Oncogene
FGF2-induced upregulation of DNA polymerase-δ p12 subunit in endothelial cells
Oncogene
Cited by (9)
Angiogenic profiling and comparison of immortalized endothelial cells for functional genomics
2008, Experimental Cell ResearchTargeted gene-delivery strategies for angiostatic cancer treatment
2007, Trends in Molecular MedicineCitation Excerpt :ECs are easily accessible and less prone to mutations compared with tumor cells; also, disrupting the vasculature by killing a few ECs results in massive tumor-cell death [3]. Furthermore, a continuously increasing number of molecules are being identified that distinguish the tumor ECs from the normal endothelium [4,5]. This makes tumor angiogenesis an excellent target for targeted gene therapy.
Theranostic nanomaterials for image-guided gene therapy
2014, MRS BulletinStructure and Functions of HMGB2 Protein
2023, International Journal of Molecular SciencesBiological functions and theranostic potential of HMGB family members in human cancers
2020, Therapeutic Advances in Medical Oncology