RESUMEN
The survival benefit of anti-vascular endothelial growth factor (VEGF) therapy in metastatic colorectal cancer (mCRC) patients is limited to a few months because of acquired resistance. We show that anti-VEGF therapy induced remodeling of the extracellular matrix with subsequent alteration of the physical properties of colorectal liver metastases. Preoperative treatment with bevacizumab in patients with colorectal liver metastases increased hyaluronic acid (HA) deposition within the tumors. Moreover, in two syngeneic mouse models of CRC metastasis in the liver, we show that anti-VEGF therapy markedly increased the expression of HA and sulfated glycosaminoglycans (sGAGs), without significantly changing collagen deposition. The density of these matrix components correlated with increased tumor stiffness after anti-VEGF therapy. Treatment-induced tumor hypoxia appeared to be the driving force for the remodeling of the extracellular matrix. In preclinical models, we show that enzymatic depletion of HA partially rescued the compromised perfusion in liver mCRCs after anti-VEGF therapy and prolonged survival in combination with anti-VEGF therapy and chemotherapy. These findings suggest that extracellular matrix components such as HA could be a potential therapeutic target for reducing physical barriers to systemic treatments in patients with mCRC who receive anti-VEGF therapy.
Asunto(s)
Bevacizumab/uso terapéutico , Neoplasias Colorrectales/patología , Neoplasias Hepáticas/secundario , Neoplasias Hepáticas/terapia , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Animales , Antineoplásicos Inmunológicos/efectos adversos , Antineoplásicos Inmunológicos/uso terapéutico , Bevacizumab/efectos adversos , Fenómenos Biomecánicos , Línea Celular Tumoral , Neoplasias Colorrectales/terapia , Resistencia a Antineoplásicos , Matriz Extracelular/patología , Matriz Extracelular/fisiología , Glicosaminoglicanos/metabolismo , Humanos , Ácido Hialurónico/metabolismo , Hipoxia/etiología , Hipoxia/fisiopatología , Neoplasias Hepáticas/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Investigación Biomédica TraslacionalRESUMEN
OBJECTIVES: It has been demonstrated that extracellular matrix molecules are involved in cyclosporine-induced gingival overgrowth (GO). However, for many of these molecules, it remains unclear whether their abundance is modulated on the protein and gene expression level. MATERIAL AND METHODS: To contribute to this clarification, we have analysed the protein and mRNA expression of type-I collagen (COL1) and decorin (DC) in native specimens obtained from five patients with GO, and matched normal tissue using indirect immunofluorescence (IIM), in situ hybridization (ISH) and quantitative polymerase chain reaction (PCR). RESULTS: IIF revealed a largely co-localized although remarkably increased abundance for COL1 and DC in GO. This increase coincided with an up-regulated gene expression observed for both molecules, as detected by ISH and quantitative PCR. CONCLUSIONS: Analysis of our data clearly demonstrates elevated levels for COL1 and DC and shows for the first time in native human tissue that involvement of these genes in GO is not confined to the protein level but also includes the transcriptional level.
Asunto(s)
Colágeno Tipo I/biosíntesis , Proteínas de la Matriz Extracelular/biosíntesis , Sobrecrecimiento Gingival/metabolismo , Proteoglicanos/biosíntesis , Adulto , Estudios de Casos y Controles , Colágeno Tipo I/genética , Ciclosporina/efectos adversos , Decorina , Proteínas de la Matriz Extracelular/genética , Femenino , Técnica del Anticuerpo Fluorescente , Expresión Génica , Sobrecrecimiento Gingival/inducido químicamente , Sobrecrecimiento Gingival/genética , Humanos , Inmunosupresores/efectos adversos , Hibridación in Situ , Trasplante de Riñón , Masculino , Persona de Mediana Edad , Proteoglicanos/genética , ARN Mensajero/análisis , Transcripción Genética , Regulación hacia Arriba , Vimentina/biosíntesisRESUMEN
In humans, pathogenesis in cyclosporine A (CsA)-induced gingival overgrowth (GO) includes the accumulation of extracellular matrix (ECM) constituents, viz., collagen type-1 and type-3 and proteoglycans, in subgingival connective tissue. However, whether this increase is associated with alterations of molecules pivotal for the turn-over of collagens and proteoglycans remains unclear. The present study explores the status of matrix metalloproteinase MMP-1 and MMP-10, which are important for fibrillar collagen and proteoglycan turn-over, and their tissue inhibitor TIMP-1, on their gene expression and protein levels in frozen sections derived from GO and matched normal tissue. In situ hybridization (ISH) revealed elevated levels of MMP-1 gene expression in the connective tissue of GO compared with normal tissue. This elevation also applied to MMP-10 and TIMP-1, the latter exhibiting the strongest gene transcription in the deep connective tissue. These differences detected by ISH were corroborated by quantitative reverse transcription/polymerase chain reaction; relative gene expression analysis indicated a 1.9-fold increase for MMP-1, a 2.3-fold increase for MMP-10, and a 4.8-fold increase for TIMP-1. Detection of the protein by indirect immunofluorescence showed that normal gingival tissue was devoid of all three proteins, although they were detectable in GO tissue, with emphasis on TIMP-1. Analysis of our data indicates elevated levels of MMP-1 and-10, and particularly TIMP-1. With respect to TIMP-1, this elevation may in turn lead to alterations in ECM turn-over by abrogating MMP-1 and MMP-10, thereby contributing to ECM accumulation associated with GO.