INTRODUÇÃO: A angiogênese tem sido proposta como um marcador de prognóstico em uma variedade de tumores incluindo o renal. No avanço da compreensão da biologia molecular do carcinoma de células renais (CCR) os genes CD105 (endoglina), VEGF, VEGFR1 e VEGFR2 são seletivamente expressos em células endoteliais vasculares sendo estudados como potenciais alvos terapêuticos. OBJETIVOS: Este estudo analisou a expressão destes quatro genes no tecido renal normal e CCR e suas correlações com os fatores prognósticos para a neoplasia. MÉTODOS: Os níveis de expressão de CD105, VEGF, VEGFR1 e VEGFR2 foram analisados por Reação da Transcriptase Reversa em tempo real (qRT-PCR) em amostras de tumor fresco congelado coletados de 56 pacientes submetidos à nefrectomia parcial ou radical por CCR. Neste estudo foram avaliados os níveis de expressão dos genes em tecidos normais e tumorais e comparados com fatores prognósticos como tamanho tumoral (> ou <= 7 cm), Grau de Fuhrman (1-2 e 3-4) e invasão microvascular (presente ou ausente). RESULTADOS: A análise dos quatro genes demonstrou que CD105 esta subexpresso em 94.7% dos casos e a superexpressão de VEGF, VEGFR1 e VEGFR2 ocorreu em 53,6%, 85,7% e 64,3% dos casos respectivamente. A expressão de endoglina foi significativamente maior em pacientes com doença metastática (p=0,05) em relação ao grupo sem metástases. Além disso, o gene do VEGFR2 foi associado com estadiamento T, apresentando uma média de expressão maior nos pacientes portadores de doença pT1-2 (p=0,04). CONCLUSÕES: Nossos experimentos demonstraram que a endoglina está subexpressa em carcinoma de células renais em relação com o tecido renal normal e a presença de níveis mais elevados está relacionada à doença metastática. Os genes VEGF, VEGFR1 e VEGFR2 encontram-se superexpressos no CCR e uma maior expressão do gene VEGFR2 está relacionada com estadiamento T1 e T2 / INTRODUCTION: Angiogenesis has been proposed as a prognostic marker in a variety of human malignancies, including renal cancer. Due to a better understanding of the underlying biology of Renal Cell Carcinoma (RCC) the genes expression of CD105 (endoglina), VEGF, VEGFR1 and VEGFR2 that are selectively expressed in vascular endothelial cells are being studied as potential therapeutic targets. OBJECTIVES: This study analyzed the expression of these four genes in normal kidney tissue and RCC and relationship with prognostic factors. METHODS: CD105, VEGF, VEGFR1 and VEGFR2 expression levels were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) in fresh-frozen malignant tissue specimens collected from 56 patients submitted to radical or partial nephrectomy. This study assessed the expression of genes in normal and tumor tissues, and compared with classic prognostic parameters in CCRCC , such as tumor size (larger or smaller than 7 cm) Fuhrman grade (1-2 and 3-4) and microvascular invasion (this or absent). RESULTS: The analysis of these four genes showed that CD105 is subexpressed in 94.7% of cases and the overexpression of VEGF, VEGFR1 and VEGFR2 occurred in 53.6%, 85.7% and 64.3% of cases respectively in tumor tissues compared to controls. The expression of endoglin was significantly higher in patients with metastatic disease (p = 0.05). In addition, VEGFR2 gene was associated with stage T, with an average of expression higher in patients with stage T1-T2 (p = 0.040). Conclusions: These experiments demonstrated that endoglin was underexpressed in RCC compared to normal kidney and the presence of enhanced expression is associated with metastatic disease. VEGF, VEGFR1 and VEGFR2 were overexpressed in the CCRCC and a higher VEGFR2 expression was related with stage T1-2

INTRODUÇÃO: A angiogênese tem sido proposta como um marcador de prognóstico em uma variedade de tumores incluindo o renal. No avanço da compreensão da biologia molecular do carcinoma de células renais (CCR) os genes CD105 (endoglina), VEGF, VEGFR1 e VEGFR2 são seletivamente expressos em células endoteliais vasculares sendo estudados como potenciais alvos terapêuticos. OBJETIVOS: Este estudo analisou a expressão destes quatro genes no tecido renal normal e CCR e suas correlações com os fatores prognósticos para a neoplasia. MÉTODOS: Os níveis de expressão de CD105, VEGF, VEGFR1 e VEGFR2 foram analisados por Reação da Transcriptase Reversa em tempo real (qRT-PCR) em amostras de tumor fresco congelado coletados de 56 pacientes submetidos à nefrectomia parcial ou radical por CCR. Neste estudo foram avaliados os níveis de expressão dos genes em tecidos normais e tumorais e comparados com fatores prognósticos como tamanho tumoral (> ou <= 7 cm), Grau de Fuhrman (1-2 e 3-4) e invasão microvascular (presente ou ausente). RESULTADOS: A análise dos quatro genes demonstrou que CD105 esta subexpresso em 94.7% dos casos e a superexpressão de VEGF, VEGFR1 e VEGFR2 ocorreu em 53,6%, 85,7% e 64,3% dos casos respectivamente. A expressão de endoglina foi significativamente maior em pacientes com doença metastática (p=0,05) em relação ao grupo sem metástases. Além disso, o gene do VEGFR2 foi associado com estadiamento T, apresentando uma média de expressão maior nos pacientes portadores de doença pT1-2 (p=0,04). CONCLUSÕES: Nossos experimentos demonstraram que a endoglina está subexpressa em carcinoma de células renais em relação com o tecido renal normal e a presença de níveis mais elevados está relacionada à doença metastática. Os genes VEGF, VEGFR1 e VEGFR2 encontram-se superexpressos no CCR e uma maior expressão do gene VEGFR2 está relacionada com estadiamento T1 e T2 / INTRODUCTION: Angiogenesis has been proposed as a prognostic marker in a variety of human malignancies, including renal cancer. Due to a better understanding of the underlying biology of Renal Cell Carcinoma (RCC) the genes expression of CD105 (endoglina), VEGF, VEGFR1 and VEGFR2 that are selectively expressed in vascular endothelial cells are being studied as potential therapeutic targets. OBJECTIVES: This study analyzed the expression of these four genes in normal kidney tissue and RCC and relationship with prognostic factors. METHODS: CD105, VEGF, VEGFR1 and VEGFR2 expression levels were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) in fresh-frozen malignant tissue specimens collected from 56 patients submitted to radical or partial nephrectomy. This study assessed the expression of genes in normal and tumor tissues, and compared with classic prognostic parameters in CCRCC , such as tumor size (larger or smaller than 7 cm) Fuhrman grade (1-2 and 3-4) and microvascular invasion (this or absent). RESULTS: The analysis of these four genes showed that CD105 is subexpressed in 94.7% of cases and the overexpression of VEGF, VEGFR1 and VEGFR2 occurred in 53.6%, 85.7% and 64.3% of cases respectively in tumor tissues compared to controls. The expression of endoglin was significantly higher in patients with metastatic disease (p = 0.05). In addition, VEGFR2 gene was associated with stage T, with an average of expression higher in patients with stage T1-T2 (p = 0.040). Conclusions: These experiments demonstrated that endoglin was underexpressed in RCC compared to normal kidney and the presence of enhanced expression is associated with metastatic disease. VEGF, VEGFR1 and VEGFR2 were overexpressed in the CCRCC and a higher VEGFR2 expression was related with stage T1-2

Pathological angiogenesis, or new blood vessel formation, is involved in many pathologies, including cancer and serious eye diseases. While traditional anti-angiogenic therapies target vascular endothelial growth factor receptors to reduce or inhibit new vessel formation, this approach has several downsides, including unpleasant side effects and low efficacy over time. Therefore, identifying new targets to treat pathological angiogenesis is still needed. CMG2, one of the two identified anthrax toxin receptors, has been proposed as an alternative target to treat pathological angiogenesis. CMG2’s role as a cell surface receptor that mediates anthrax toxin internalization is very well documented. One physiological function for CMG2, not related to anthrax intoxication, is suggested by the observation that loss-of-function mutations in CMG2 cause hyaline fibromatosis syndrome (HFS), a genetic disease that results in accumulations of extra-cellular matrix (ECM) protein in different parts of the body. While the complete molecular mechanism for CMG2’s role in regulating angiogenesis has not been determined, this dissertation addresses multiple ways CMG2 regulates pathological angiogenesis. We have discovered that CMG2 plays a role in mediating ECM homeostasis via endocytosis of ECM proteins and protein fragments as a way to generate angiogenic signals from the cell. We have also demonstrated that a fragment from Col IV, S16, is endocytosed into the cells by interacting with CMG2, and S16 treatment to endothelial cells leads to a significant reduction in cell migration. Also, an endothelial cell migration assay with CMG2 knockout cells results in abolished directional migration, indicating that CMG2 is required for endothelial cell chemotaxis. Notably, we have identified that bFGF, VEGF, and PDGF are involved in CMG2 mediated chemotaxis but not insulin and sphingosine-1-phosphate (S1P). While recent literature reports show that CMG2 works closely with RhoA GTPase, which is commonly known to regulate cell migration, we have also observed that inhibition of RhoA also reduced cell chemotaxis towards VEGF but not S1P. These results could be leveraged to develop new classes of therapeutic molecules to treat pathological angiogenesis induced by multiple various growth factors via targeting CMG2.

Ischemic retinopathies (IR) are vision-threatening diseases that affect a substantial amount of people across all age groups worldwide. The current treatment options of photocoagulation and anti-VEGF therapy have side effects and are occasionally unable to prevent disease progression. It is therefore worthwhile to consider other molecular targets for the development of novel treatment strategies that could be safer and more efficient. During the manifestation of IR, the retina, normally an immune privileged tissue, encounters enhanced levels of cellular stress and inflammation that attract mononuclear phagocytes (MPs) from the blood stream and activate resident MPs (microglia). Activated MPs have a multitude of effects within the retinal tissue and have the potential to both counter and exacerbate the harmful tissue microenvironment. The present review discusses the current knowledge about the role of inflammation and activated retinal MPs in the major IRs: retinopathy of prematurity and diabetic retinopathy. We focus particularly on MPs and their secreted factors and cell–cell-based interactions between MPs and endothelial cells. We conclude that activated MPs play a major role in the manifestation and progression of IRs and could therefore become a promising new target for novel pharmacological intervention strategies in these diseases.