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Angiogenesis--a new target for future therapy.

Development of blood vessels from in situ differentiating endothelial cells (EC) is called vasculogenesis, whereas sprouting of new blood vessels from the pre-existing ones is termed angiogenesis or neovascularisation. Angiogenesis, the growth of new blood vessels, is essential during tissue repair, foetal development, and female reproductive cycle. In contrast, uncontrolled angiogenesis promotes tumor and retinopathies, while inadequate angiogenesis can lead to coronary artery disease. A balance between pro-angiogenic and anti-angiogenic growth factors and cytokines tightly controls angiogenesis. With the identification of several proangiogenic molecules such as the vascular endothelial cell growth factor (VEGF), the fibroblast growth factors (FGFs), and the angiopoietins, and the recent description of specific inhibitors of angiogenesis such as platelet factor-4, angiostatin, endostatin, and vasostatin, it is recognized that therapeutic interference with vasculature formation offers a tool for clinical applications in various pathologies. Inhibition of angiogenesis can prevent diseases such as cancer, diabetic nephropathy, arthritis, psoriasis, whereas stimulation of angiogenesis is beneficial in the treatment of coronary artery disease (CAD), cardiac failure, tissue injury, etc. One of the most specific and critical regulators of angiogenesis is vascular endothelial growth factor (VEGF), which regulates endothelial proliferation, permeability, and survival. Substantial evidence also implicates VEGF as an angiogenic mediator in tumors and intraocular neovascular syndromes, and numerous clinical trials are presently testing the hypothesis that inhibition of VEGF may have therapeutic value.

Adiponectin↗

Advances in angiogenesis research: relevance to urological oncology.

PURPOSE: Important advances in angiogenesis research are reviewed along with recent data implicating angiogenesis in the pathogenesis of urological malignancies. MATERIALS AND METHODS: The current understanding of angiogenesis and its importance in tumor biology is summarized. The rationale for anti-angiogenic therapy is reviewed and the clinical experience with these agents is discussed. An extensive literature search of angiogenesis in urological malignancies was performed. RESULTS: Quantitative immunohistochemistry for endothelial antigens suggests that, as is the case with many other tumors, induction of angiogenesis contributes to the malignant phenotype of prostate and bladder carcinomas. Anti-angiogenic agents have demonstrated efficacy against urological tumors in experimental systems, and recent data suggest that these agents may also be useful for chemoprophylactic purposes. Putative angiogenesis inducers specific for each of the major urological malignancies have been identified. Quantitation of the expression of angiogenesis inducers and estimation of microvessel density have demonstrated prognostic value for urological malignancies. CONCLUSIONS: The available data indicate that angiogenesis has an important role in the progression and metastasis of urological malignancies. Preclinical data coupled with experience in other cancers indicate that combining anti-angiogenic therapy with conventional treatment modalities has the potential to improve dramatically our management of these malignancies. Further research will be needed to define the mechanisms controlling angiogenesis in urological malignancies and to determine if any of the angiogenic correlates will be of genuine clinical use. The rapid pace of research in this field suggests that this aspect of tumor biology will soon have an increasingly important role in the evaluation and treatment of urological cancers.

Angiogenesis Inducing Agents↗

Matrix metalloproteinases and angiogenesis.

Matrix metalloproteinases (MMPs) are a family of enzymes that proteolytically degrade various components of the extracellular matrix (ECM). Angiogenesis is the process of forming new blood vessels from existing ones and requires degradation of the vascular basement membrane and remodeling of the ECM in order to allow endothelial cells to migrate and invade into the surrounding tissue. MMPs participate in this remodeling of basement membranes and ECM. However, it has become clear that MMPs contribute more to angiogenesis than just degrading ECM components. Specific MMPs have been shown to enhance angiogenesis by helping to detach pericytes from vessels undergoing angiogenesis, by releasing ECM-bound angiogenic growth factors, by exposing cryptic proangiogenic integrin binding sites in the ECM, by generating promigratory ECM component fragments, and by cleaving endothelial cell-cell adhesions. MMPs can also contribute negatively to angiogenesis through the generation of endogenous angiogenesis inhibitors by proteolytic cleavage of certain collagen chains and plasminogen and by modulating cell receptor signaling by cleaving off their ligand-binding domains. A number of inhibitors of MMPs that show antiangiogenic activity are already in early stages of clinical trials, primarily to treat cancer and cancer-associated angiogenesis. However, because of the multiple effects of MMPs on angiogenesis, careful testing of these MMP inhibitors is necessary to show that these compounds do not actually enhance angiogenesis.

Angiogenesis Inhibitors↗

[Angiogenesis in gynecology and obstetrics].

In current scientific discussion, increasing importance is being given to the clinical significance of the new formation of vessels (angiogenesis) in the course of physiological, inflammatory and neoplastic processes. Angiogenesis is best studied in the growth of malignant tumors, since cancer may be regarded as the most important angiogenesis-dependent disease in terms of social and economic aspects. The significance of angiogenesis in gynecological oncology is as follows: 1) Intratumoral vessel density is an indicator for the emergence and growth of malignant tumours and their precursor lesions, 2) intratumoral vessel density is an independent prognostic factor for solid malignancies and 3) the inhibition of tumor angiogenesis by means of anti-angiogenetic substances causes tumor growth to be suppressed. Angiogenesis also plays an important role in the regulation of the female menstrual cycle. Proliferation of the endometrium and the formation of the corpus luteum in the second half of the menstrual cycle are examples of angiogenesis in the physiological field. The function of angiogenetic factors in the emergence of endometriosis and in female and male infertility are currently under study. In obstetrics, the new formation of blood vessels is significant for the implantation of impregnated blastocysts and for the development and growth of the placenta. Preeclempsia (gestational toxicosis), for instance, is a typical pregnancy-related disease whose pathophysiological mechanism is attributed to a disturbed development and function of small placental vessels. The present paper is an overview of current knowledge and current approaches of research concerning angiogenesis in the field of gynecology and obstetrics. The paper is focused on the clinical significance of angiogenesis.

Angiogenesis Inducing Agents↗

Understanding angiogenesis and its clinical applications.

Angiogenesis is the growth of new vessels from pre-existing blood vessels. Angiogenesis is critical during embryogenesis but occurs minimally in healthy adults, except in wound repair, inflammation, female reproductive organs, and pathologic conditions. Various growth factors and proteins, elements of the extracellular matrix, components of the coagulation/fibrinolytic system, and platelets interact with the endothelial cells and pericytes of blood vessels to regulate angiogenesis. Characterization of angiogenic factors has revealed that remodeling of the extracellular matrix occurs during angiogenesis, mediated by integrins that are found on the endothelial cell surface membrane. Counter-regulatory antiangiogenic proteins and molecules that show an intricate balance in the regulation of angiogenesis have also been characterized. Components of the coagulation/fibrinolysis cascade also play a critical role in angiogenesis. Elucidation of the mechanisms of angiogenesis has led to better understanding of certain disease states. Ongoing studies are evaluating the stimulation of angiogenesis to treat ischemic disorders, and the inhibition of angiogenesis to prevent abnormal proliferation in malignant and non-malignant disorders.

Adult↗

VEGF, bFGF and EGF in the angiogenesis of human melanoma xenografts.

Vascular endothelial growth factor (VEGF) is a major inducer of angiogenesis in tumors. Basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) have both been shown to interact with VEGF. The involvement of VEGF, bFGF, and EGF in melanoma angiogenesis was investigated here. Four human melanoma cell lines (A-07, D-12, R-18, U-25) were included in the study. Angiogenesis was quantified by scoring of tumor-oriented capillaries following intradermal cell inoculation in BALB/c nu/nu mice. VEGF, bFGF and EGF expression and secretion were investigated by Western blotting and enzyme-linked immunosorbent assay, respectively. Immunohistochemistry of xenografts grown intradermally was used to reveal VEGF and bFGF localization in vivo. The rate of angiogenesis differed substantially among the melanoma lines; the sequence from a high to low rate of angiogenesis was: A-07, D-12, R-18, U-25. A-07, which induced the highest rate of angiogenesis, showed a higher rate of VEGF secretion, stronger VEGF staining by immunohistochemistry and higher bFGF expression than the other lines. U-25, which induced the lowest rate of angiogenesis, showed a higher rate of VEGF secretion than D-12 and R-18. A-07 was the only line that showed detectable bFGF secretion, and R-18 was the only line that showed detectable EGF secretion. VEGF is probably important in the angiogenesis of melanomas. However, heterogeneity in rate of angiogenesis among melanomas cannot be attributed to heterogeneity in rate of secretion of VEGF, bFGF and/or EGF only.

Animals↗

Prognostic value of bone marrow angiogenesis in multiple myeloma: use of light microscopy as well as computerized image analyzer in the assessment of microvessel density and total vascular area in multiple myeloma and its correlation with various clinical, histological, and laboratory parameters.

We studied the prognostic value of parameters of angiogenesis on bone marrow biopsies in newly diagnosed multiple myeloma (MM) patients. Angiogenesis parameters studied were the microvessel count done manually on light microscopy (MVD-A), microvessel count done by using computerized image analyzer (MVD-B), and total vascular area (TVA) measured by computerized image analyzer. One hundred ten newly diagnosed cases of MM treated at Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, were analyzed with respect to clinical features, laboratory findings, histological features, angiogenesis parameters, and responses to the treatment on follow-up. Twenty age- and sex-matched controls were studied for comparing with angiogenesis of the test cases. Bone marrow microvessels were examined using immunohistochemical staining for CD34. MVD-A (range 4.9-85.2; mean 28.2; SD 19.4), MVD-B (range 2.0-26.9; mean 11.7; SD 5.9), and TVA measured in percentage (range 0.1-17.1; mean 2.4; SD 2.5) were measured for test cases (n = 110). Grading of angiogenesis parameters of the test cases were done; such that angiogenesis parameters of controls (taken as baseline) were grade I. There was a statistically highly significant correlation between (MVD-A vs MVD-B, pcc = 0.92; MVD-A vs TVA, pcc = 0.78; MVD-B vs TVA, pcc = 0.76). The myeloma cases had significantly higher angiogenesis parameters when compared with controls (Kruskall-Wallis test, P < 0.001). "Complete responders" (n = 38/110) had significant lower angiogenesis (Mann-Whitney U test, P < 0.001) than "nonresponders" (n = 72/110). On treatment follow-up "rapid disease progressors" had the highest levels of angiogenesis (mean rank for MVD-A = 84.7, MVD-B = 82.1, and TVA = 81.1). On multivariate (logistic regression) analysis, factors found to have independent prognostic significance in complete responders (adjusted odd ratio (95% CI, P value)] were: (a) MVD-B grade I [0.134 (0.10-0.16, P < 0.001)], (b) clinical substage A [0.163 (0.12-0.19, P = 0.008)], (c) Bartl's histological stage II & I [0.262 (0.2-0.32, P = 0.021)], (d) MVD-A grade I [0.28 (0.22-0.36, P = 0.03)], (e) beta2 microglobulin levels less than 3,400 ng/dl [0.31 (0.23-0.42, P = 0.04)]. Kaplan-Meier survival analysis for myeloma-related death (n = 16) shows a mean survival time (in months) of 24.75; SE = 3; 95% CI = 21-28. We conclude that MVD (particularly MVD-B) is a very good predictor for the complete response in patients of MM and should be done routinely on bone marrow biopsies.

Adult↗

From the cradle to the clinic: VEGF in developmental, physiological, and pathological angiogenesis.

Formation of new blood vessels, which is fundamental in embryonic development, occurs through a combination of angiogenesis and vasculogenesis. Angiogenesis also plays a vital role postnatally, especially in reparative processes such as wound and fracture healing. Some of these events, especially in fracture healing, recapitulate processes observed in developmental angiogenesis. However, dysregulated angiogenesis is well documented to underlie a number of pathological disorders, including rheumatoid arthritis (RA). The vascular endothelial growth factor (VEGF)/VEGF receptor system is the best characterized regulator of angiogenesis. VEGF is expressed in a range of cells in response to soluble mediators (such as cytokines and growth factors), cell-bound stimuli (such as CD40 ligand), and environmental factors (such as hypoxia). As a consequence, this molecule is vital in the modulation of physiological and pathological angiogenesis. This review will focus in particular on the role played by VEGF in embryogenesis and skeletal growth, in fracture healing (in which increased angiogenesis is likely to be beneficial in promoting union), and in RA (in which excessive angiogenesis is thought to play a significant role in disease pathogenesis). In the not-too-distant future, targeting VEGF may prove to be of benefit in the treatment of diseases associated with excessive or aberrant angiogenesis, such as malignancies and RA.

Arthritis, Rheumatoid↗

Angiogenesis in malignancies of the female genital tract.

OBJECTIVE: The purpose of this work was to review current knowledge pertaining to angiogenesis in malignancies of the female genital tract. METHODS: We identified studies published in the English language regarding angiogenesis in gynecologic malignancies. The studies were obtained from a MEDLINE search from 1966 through June 1998; additional sources were identified through cross-referencing. RESULTS: A growing body of evidence confirms the ability of vulvar and cervical squamous cell carcinomas and endometrial and ovarian adenocarcinoma to induce angiogenesis. In vulvar intraepithelial neoplasia a correlation between vascular endothelial growth factor (VEGF) expression, microvessel density (MVD), and progression of dysplasia has been demonstrated. In invasive vulvar carcinoma, high VEGF expression and MVD portend poor prognosis. Currently a debate exists regarding the ability of cervical squamous intraepithelial neoplasia to induce angiogenesis. Most studies, however, indicate angiogenesis to be of prognostic value in patients with invasive squamous cell carcinoma. The ability of complex endometrial hyperplasia to induce angiogenesis has been demonstrated. A direct correlation between angiogenesis, higher grade and depth of invasion in Stage I adenocarcinoma, and prognostic value in Stage I and II and recurrent disease has been noted. In ovarian epithelial adenocarcinoma, higher microvessel counts in the primary ovarian tumor or omental metastases may serve as a prognostic indicator for survival. CONCLUSIONS: Similar to other malignant diseases, angiogenesis appears to play an important role in disease progression and survival in patients with gynecologic malignancies. Preliminary data indicate angiogenesis may serve as a prognostic indicator in vulvar and cervical squamous cell carcinomas and endometrial and ovarian adenocarcinomas. These findings may lead to future application of therapeutic trials with antiangiogenic factors.

Adenocarcinoma↗

Regional differences in spontaneously occurring angiogenesis in the adult rat mesentery.

Despite intensive study in the area of angiogenesis, relatively little is known about normal angiogenesis in adult animals. Preliminary studies using the Griffonia simplicifolia I (GSI) lectin as a microvascular marker indicated that capillary sprouting occurs in the clear "windows" of normal intact adult rat mesentery. The purpose of this study was to determine whether angiogenesis occurs uniformly within the mesenteric windows and whether maturational age affects the extent of angiogenesis in the absence of any experimental or pathological perturbation. Four groups of female Sprague-Dawley rats were used, "weanling" (4-5 weeks), "juvenile" (6-8 weeks), "young adult" (10-13 weeks), and "adult" (16-20 weeks). Microvessels sprouting into proximal and distal windows were delineated in whole mounts by use of fluorescent derivative of the GSI lectin. Microvascular sprouting, indicating angiogenesis, was found in all age groups, but was most frequent in the windows sampled from the distal region of the small intestine when compared with those from the proximal region. The mean number of microvessels per sample site was significantly higher in the distal windows of adults than in the weanling or juvenile rats. Angiogenesis was found to occur asymmetrically within the individual windows in the two adult groups, with significantly more angiogenesis on the intestinal side compared to that along the portal vessels. We conclude that there is an age-related increase in angiogenesis into the mesenteric windows, and that the intestinal side is more prone to spontaneous angiogenesis than is the portal side.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Increased ovarian follicular angiogenesis and dynamic changes of follicular vascular plexuses induced by equine chorionic gonadotropin in the gilt.

Follicular angiogenesis and capillary degeneration are crucial ovarian processes in folliculogenesis. The present study was conducted to assess the changes in population of follicular vascular plexuses with different capillary status in prepubertal gilts 72 h after equine chorionic gonadotropin (eCG) (1,250 IU) treatment, using combined vascular corrosion casting and scanning electron microscopy. Follicular fluid concentrations of estradiol, progesterone and vascular endothelial growth factor (VEGF) were determined to confirm the follicular status. Based on the proliferative or degenerative characteristics of their capillaries, follicles were classified into three categories: active angiogenesis, low angiogenesis and degeneration. Irrespective of exogenous gonadotropin treatment in vivo, small follicular vascular plexuses (<4 mm in diameter) exhibited all three conditions in casted ovaries, while medium (4-5 mm) and large (>5 mm) plexuses showed only active angiogenesis or degeneration. eCG treatment significantly increased the population of large, but decreased that of small follicular plexuses. Most large follicular vascular plexuses showed active angiogenesis with higher follicular fluid estradiol: progesterone ratios and VEGF concentration. eCG also increased the percentage of medium follicular plexuses with active angiogenesis. The populations of small follicular plexuses with active angiogenesis were higher in controls, but decreased after eCG treatment. However, treatment of gilts with the gonadotropin increased the percentage of small plexuses (<1.0 mm) with low angiogenesis and those (1-3.9 mm) with extensive capillary degeneration. These findings are consistent with the hypothesis that angiogenesis is involved in selection and growth of small follicles in gilts under the regulation of gonadotropin.

Animals↗

Immunostimulatory DNA inhibits allergen-induced peribronchial angiogenesis in mice.

BACKGROUND: Airway remodeling in asthma is associated with angiogenesis. OBJECTIVE: We have examined whether immunostimulatory sequences of DNA (ISSs) inhibit allergen-induced airway angiogenesis and expression of angiogenic cytokines in a mouse model of airway remodeling. METHODS: Mice sensitized to ovalbumin were challenged repetitively with ovalbumin for three months to develop airway remodeling and angiogenesis. Levels of angiogenesis were compared in ISS-treated and control mice. RESULTS: Mice challenged with ovalbumin developed significantly increased levels of peribronchial angiogenesis (increase in the number of CD31+ peribronchial small blood vessels) and an increase in the peribronchial vascular area as assessed by image analysis. Ovalbumin-induced peribronchial angiogenesis was associated with increased bronchoalveolar lavage levels of vascular endothelial growth factor (VEGF) and an increase in the number of peribronchial cells expressing VEGF. Treatment of mice with ISS before repetitive ovalbumin challenge significantly reduced the levels of peribronchial angiogenesis as well as the levels of bronchoalveolar lavage VEGF and the number of peribronchial cells expressing VEGF. ISS is unlikely to act directly on endothelial cells to inhibit angiogenesis because lung endothelial cells did not express Toll receptor 9, the receptor for ISS as assessed by RT-PCR. In vitro studies demonstrated that ISS inhibited macrophage expression of VEGF. CONCLUSION: The ability of ISS to inhibit angiogenesis in vivo is likely to be mediated by several mechanisms, including ISS reducing the number of peribronchial inflammatory cells that express VEGF, ISS inhibiting expression of TH2 cytokines such as IL-13 that promote VEGF expression, and direct effects of ISS on macrophages to inhibit VEGF expression.

Adjuvants, Immunologic↗

Effect of thymosin peptides on the chick chorioallantoic membrane angiogenesis model.

The effect of alpha- and beta-thymosin peptides, namely prothymosin alpha (ProT(alpha)), thymosin alpha(1) (T(alpha)1), parathymosin alpha (ParaT(alpha)), thymosin beta(4) (Tbeta4), thymosin beta(10) (Tbeta10), and thymosin beta(9) (Tbeta9), on the angiogenesis process was investigated using the chick chorioallantoic membrane as an in vivo angiogenesis model. The thymosin peptides tested were applied in 10 microl aliquots containing 0.01-4 nmoles of Tbeta4, Tbeta10 or Tbeta9, 0.016-6.66 nmoles of T(alpha)1, 4.1 pmoles-1.66 nmoles of ProT(alpha), and 4.4 pmoles-1.76 nmoles of ParaT(alpha). Phorbol 12-myristate 13-acetate and hydrocortisone were also used as positive and negative control, respectively. Tbeta4, ProT(alpha) and T(alpha)1 were found to enhance angiogenesis, while Tbeta10, Tbeta9 and ParaT(alpha) exhibited an inhibitory effect on the angiogenesis process. When mixtures of Tbeta4 and Tbeta10 containing active amounts of the two peptides at different proportions were applied, the promoting effect of Tbeta4 on angiogenesis was reversed in the presence of increasing concentrations of Tbeta10 and vice versa. The effect of Tbeta10, Tbeta9, ProT(alpha) and ParaT(alpha), in parallel with Tbeta4 and T(alpha)1, on the angiogenesis process was investigated for the first time as far as we know and the results of this study offer more insight into the biological regulatory roles of thymosin peptides, and provide helpful information about their therapeutic potential. Whether these agents could be used either as inhibitors of angiogenesis in disease states where uncontrolled angiogenesis is involved, e.g. in carcinogenesis, or as angiogenesis promoters that could be useful in wound healing, fracture repair, peptic ulcers etc., remains to be further studied.

Allantois↗

Reactive oxygen species as mediators of angiogenesis signaling: role of NAD(P)H oxidase.

Angiogenesis, a process of new blood vessel growth, contributes to various pathophysiologies such as cancer, diabetic retinopathy and atherosclerosis. Accumulating evidence suggests that cardiovascular diseases are associated with increased oxidative stress in blood vessels. Reactive oxygen species (ROS) such as superoxide and H2O2 cause blood vessels to thicken, produce inflammation in the vessel wall, and thus are regarded as "risk factors" for vascular disease, whereas ROS also act as signaling molecules in many aspects of growth factor-mediated physiological responses. Recent reports suggest that ROS play an important role in angiogenesis; however, its underlying molecular mechanisms remain unknown. Vascular endothelial growth factor (VEGF) induces angiogenesis by stimulating endothelial cell (EC) proliferation and migration primarily through the receptor tyrosine kinase VEGF receptor2 (Flk1/KDR). VEGF binding initiates tyrosine phosphorylation of KDR, which results in activation of downstream signaling enzymes including ERK1/2, Akt and eNOS, which contribute to angiogenic-related responses in EC. Importantly, the major source of ROS in EC is a NAD(P)H oxidase and EC express all the components of phagocytic NAD(P)H oxidase including gp91phox, p22phox, p47phox, p67phox and the small G protein Rac1. We have recently demonstrated that ROS derived from NAD(P)H oxidase are critically important for VEGF signaling in vitro and angiogenesis in vivo. Furthermore, a peptide hormone, angiotensin II, a major stimulus for vascular NAD(P)H oxidase, also plays an important role in angiogenesis. Because EC migration and proliferation are primary features of the process of myocardial angiogenesis, we would like to focus on the recent progress that has been made in the emerging area of NAD(P)H oxidase-derived ROS-dependent signaling in ECs, and discuss the possible roles in angiogenesis. Understanding these mechanisms may provide insight into the components of NAD(P)H oxidase as potential therapeutic targets for treatment of angiogenesis-dependent diseases such as cancer and atherosclerosis and for promoting myocardial angiogenesis in ischemic heart diseases.

Angiotensins↗

Basement membrane biosynthesis as a target for developing inhibitors of angiogenesis with anti-tumor properties.

Basement membrane (BM) exerts profound influence on endothelial cell (EC) behavior. In addition BM is a structural element of blood vessels; in fact at some point of their formation blood vessels are bare EC tubes lined with the BM produced by these EC. We thought, therefore, that a quantitative relationship must exist between the rate of BM synthesis and angiogenesis, and that interfering with BM synthesis must have an effect on angiogenesis. This was found experimentally in the chick chorioallantoic membrane (CAM) system. It was shown that the rate of BM collagen biosynthesis can serve as a biochemical index of angiogenesis and that inhibitors of BM synthesis prevent angiogenesis. GPA 1734 (8,9-dihydroxy-70-methyl-benzo(b)quinolizinium bromide), which inhibits proline and lysine hydroxylations in type IV collagen formation, suppresses angiogenesis in the CAM. Similarly, D609 (tricyclodecan-9-yl-xanthate), which inhibits BM synthesis by an as yet unknown mechanism, also prevents angiogenesis. Structurally related analogs of GPA 1734 and D609 that have no effect on BM biosynthesis are also without effect on angiogenesis. The aforementioned inhibitors of angiogenesis GPA 1734 and D609 have a dose-dependent inhibitory effect on tumor growth in rats bearing Walker 256 carcinosarcoma, without any obvious toxic effects. This effect is probably related to angiosuppression, since structurally related analogs that do not inhibit angiogenesis are without antitumor properties. Also GPA 1734 and D609 have no direct cytotoxic effects on Walker 256 cells in vitro. These results suggest that a search for agents that are specific inhibitors of BM synthesis may provide novel angiosuppressors with potential application in tumor chemotherapy.(ABSTRACT TRUNCATED AT 250 WORDS)

2H-Benzo(a)quinolizin-2-ol, 2-Ethyl-1,3,4,6,7,11b-↗

Suppression of experimental corneal angiogenesis by focal X-ray irradiation.

PURPOSE: To investigate the effect of focal X-ray irradiation on experimental corneal angiogenesis in the rabbit. METHODS: A gelatin hydrogel sheet impregnated with basic fibroblast growth factor was implanted into the corneal stroma of rabbits; this induced corneal angiogenesis. After the first sign of corneal angiogenesis was noted, the corneal region was irradiated with a dose of 10 Gy or 20 Gy. The control rabbits received no irradiation. The eyes were examined by slitlamp biomicroscopy and photographed, over a period of 28 days. The maximum length and total surface area of corneal angiogenesis were quantified by computerized image analysis. RESULTS: Corneal angiogenesis was noted on day 3 following implantation of the hydrogel sheet. In the rabbits irradiated with 10 Gy, the maximum length and total surface area of corneal angiogenesis were both significantly lower on day 4 and 7 following irradiation, compared to the respective measurement in the controls. In the rabbits irradiated with 20 Gy, the maximum length and total surface area of corneal angiogenesis were significantly lower between days 4 and 21, and between days 4 and 14, respectively, compared to the respective measurement in the controls. CONCLUSIONS: Focal X-ray irradiation to the corneal region suppressed corneal angiogenesis in a dose-dependent manner. Focal X-ray irradiation may be beneficial in treating ocular angiogenesis.

Animals↗

Involvement of cysteine proteases in bFGF-induced angiogenesis in guinea pig and rat cornea.

Overexpression and activation of matrix metalloprotease (MMP) have been implicated in angiogenesis. However, the involvement of cysteine proteases, such as calpains (EC 34.22.17), is obscure. Thus, the purpose of this experiment was to study the involvement of cysteine proteases in angiogenesis induced by basic fibroblast growth factor (bFGF) in guinea pig and rat corneas using cysteine protease inhibitors. Sustained-release polymers containing bFGF were implanted into guinea pig and rat corneas to induce angiogenesis. For treatment of corneal angiogenesis, polymers containing cysteine protease inhibitors, leupeptin or SJA6017, were also implanted into corneas. Using the slit lamp, the corneas were observed for nine days after polymer implantation. Soluble proteins and albumin levels were used as markers of corneal injury by angiogenesis. bFGF induced angiogenesis in guinea pig and rat corneas. In guinea pig cornea, wet weight, the amount of soluble protein and albumin was highest at four days after bFGF-containing pellet implantation. In rat cornea, the amount of soluble protein and albumin was highest at six days, and wet weight increased within four days. One hundred nmole of leupeptin showed a tendency to reduce bFGF-induced angiogenesis in guinea pig cornea, and 10 nmole of SJA6017 was effective in reducing bFGF-induced angiogenesis in rat cornea, although SJA6017 showed a stronger effect than leupeptin. Ten nmole of SJA6017 significantly reduced the number of new blood vessels. These data suggested involvement of cysteine proteases in angiogenesis in guinea pig and rat cornea.

Albumins↗

COX-2/VEGF-dependent facilitation of tumor-associated angiogenesis and tumor growth in vivo.

Nonsteroidal anti-inflammatory drugs are known to suppress the occurrence and progression of malignancies such as colorectal cancers. However, the precise mechanism of these actions remains unknown. We have evaluated the role of an inducible cyclo-oxygenase (COX-2) in tumor-associated angiogenesis and tumor growth, and identified the downstream molecules involved using a ddy mouse model of sponge angiogenesis, which mimics tumor angiogenesis and is COX-2 and vascular endothelial growth factor (VEGF) dependent. In this model, VEGF expression was down-regulated by selective COX-2 inhibition with NS-398. To find out the involvement of COX-2/VEGF pathway in tumor-associated angiogenesis, we estimated angiogenesis occurring around implanted Millipore chambers containing sarcoma-180 (S-180) cells or Lewis lung carcinoma cells. Daily oral administration of NS-398 or of aspirin, a nonselective COX inhibitor, suppressed angiogenesis seen around the Millipore chambers. S-180 cells implanted in ddy mice formed substantial tumors with extensive angiogenesis markedly suppressed by aspirin and COX-2 inhibitors NS-398 and JTE522, but not by mofezolac, an inhibitor of constitutive COX-1. Tumor-associated angiogenesis was also significantly suppressed by a neutralizing antibody against VEGF. S-180 tumor growth in the subcutaneous tissues was also suppressed by aspirin, COX-2 selective inhibitors, and the VEGF antibody, but not by the COX-1 inhibitor. These results demonstrate that the inhibition of the COX-2/VEGF-dependent pathway was effective in tumor-associated angiogenesis, tumor growth, and tumor metastasis.

Animals↗