Haematopoietic focal adhesion kinase deficiency alters haematopoietic homeostasis to drive tumour metastasis

Metastasis is the main cause of cancer-related death and thus understanding the molecular and cellular mechanisms underlying this process is critical. Here, our data demonstrate, contrary to established dogma, that loss of haematopoietic-derived focal adhesion kinase (FAK) is sufficient to enhance tumour metastasis. Using both experimental and spontaneous metastasis models, we show that genetic ablation of haematopoietic FAK does not affect primary tumour growth but enhances the incidence of metastasis significantly. At a molecular level, haematopoietic FAK deletion results in an increase in PU-1 levels and decrease in GATA-1 levels causing a shift of hematopoietic homeostasis towards a myeloid commitment. The subsequent increase in circulating granulocyte number, with an increase in serum CXCL12 and granulocyte CXCR4 levels, was required for augmented metastasis in mice lacking haematopoietic FAK. Overall our findings provide a mechanism by which haematopoietic FAK controls cancer metastasis

Efeito da Analgesia Preventiva na Redução da Dor Pós-Operatória: Uma Análise Abrangente

This article explores the significant intersection between anesthesia and chronic pain therapy in cancer patients, highlighting the importance of these approaches in the context of treating various neoplastic conditions. Anesthesia plays a crucial role in surgical procedures and therapeutic interventions, requiring a comprehensive analysis of available options to ensure the safety and comfort of patients. When considering different anesthetic techniques, the goal is to provide healthcare professionals with an in-depth understanding of the latest and most effective strategies, taking into account factors such as postoperative recovery and acute pain management. The choice of anesthetic approach in cancer patients is multifaceted, influenced by variables such as the type of intervention, the patient's overall health, and the specificities of the neoplastic condition. In this context, the comparative analysis of different anesthetic techniques, such as regional and general approaches, becomes essential to ensure treatment effectiveness and minimize adverse impacts. Furthermore, chronic pain therapy in cancer patients is a complex challenge that requires an integrated approach. This article examines various therapeutic modalities, from pharmacological interventions to non-pharmacological methods, with the aim of providing valuable insights to optimize chronic pain management and improve post-treatment quality of life. Understanding the complexities and nuances of these therapies is essential to ensure comprehensive and personalized care for cancer patients, emphasizing the importance of a holistic approach in managing chronic pain in an oncological context. Finally, this article seeks to consolidate up-to-date information on anesthesia and chronic pain therapy in cancer patients, offering a critical review of available scientific literature. The goal is to contribute to the continuous development of clinical practices in this area, promoting the effectiveness of procedures and improving the quality of life for patients facing this challenging condition.Este artigo explora a significativa interseção entre a anestesia e a terapia de dor crônica em pacientes oncológicos, destacando a importância dessas abordagens no contexto do tratamento de diversas condições neoplásicas. A anestesia desempenha um papel crucial em procedimentos cirúrgicos e intervenções terapêuticas, exigindo uma análise abrangente das opções disponíveis para garantir a segurança e o conforto dos pacientes. Ao considerar diferentes técnicas anestésicas, o objetivo é fornecer aos profissionais de saúde uma visão aprofundada das estratégias mais recentes e eficazes, levando em conta fatores como a recuperação pós-operatória e o manejo da dor aguda. A escolha da abordagem anestésica em pacientes oncológicos é multifacetada, influenciada por variáveis como o tipo de intervenção, estado de saúde geral do paciente e especificidades da condição neoplásica. Neste contexto, a análise comparativa de diferentes técnicas anestésicas, como abordagens regionais e gerais, torna-se essencial para garantir a eficácia do tratamento e minimizar os impactos adversos. Além disso, a terapia de dor crônica em pacientes oncológicos é um desafio complexo que requer uma abordagem integrada. Este artigo examina diversas modalidades terapêuticas, desde intervenções farmacológicas até métodos não farmacológicos, com o intuito de fornecer insights valiosos para otimizar o manejo da dor crônica e melhorar a qualidade de vida pós-tratamento. A compreensão das complexidades e nuances dessas terapias é essencial para garantir um cuidado abrangente e personalizado aos pacientes oncológicos, destacando a importância de uma abordagem holística na gestão da dor crônica em contexto oncológico. Por fim, este artigo busca consolidar informações atualizadas sobre anestesia e terapia de dor crônica em pacientes oncológicos, oferecendo uma revisão crítica da literatura científica disponível. O objetivo é contribuir para o desenvolvimento contínuo das práticas clínicas nessa área, promovendo a eficácia dos procedimentos e a melhoria da qualidade de vida para os pacientes que enfrentam essa condição desafiadora. &nbsp

Use of the mouse aortic ring assay to study angiogenesis

Here we provide a protocol for quantitative three-dimensional ex vivo mouse aortic ring angiogenesis assays, in which developing microvessels undergo many key features of angiogenesis over a timescale similar to that observed in vivo. The aortic ring assay allows analysis of cellular proliferation, migration, tube formation, microvessel branching, perivascular recruitment and remodeling-all without the need for cellular dissociation-thus providing a more complete picture of angiogenic processes compared with traditional cell-based assays. Our protocol can be applied to aortic rings from embryonic stage E18 through to adulthood and can incorporate genetic manipulation, treatment with growth factors, drugs or siRNA. This robust assay allows assessment of the salient steps in angiogenesis and quantification of the developing microvessels, and it can be used to identify new modulators of angiogenesis. The assay takes 6-14 d to complete, depending on the age of the mice, treatments applied and whether immunostaining is performed

avβ3 Integrin Limits the Contribution of Neuropilin-1 to Vascular Endothelial Growth Factor-induced Angiogenesis

Both vascular endothelial growth factor receptors (VEGFR) and integrins are major regulators of VEGF-induced angiogenesis. Previous work has shown that ß3 integrin can regulate negatively VEGFR2 expression. Here we show that ß3 integrin can regulate negatively VEGF-mediated angiogenesis by limiting the interaction of the co-receptor NRP1 (neuropilin-1) with VEGFR2. In the presence of avß3 integrin, NRP1 contributed minimally to VEGF-induced angiogenic processes in vivo, ex vivo, and in vitro. Conversely, when ß3 integrin expression is absent or low or its function is blocked with RGD-mimetic inhibitors, VEGF-mediated responses became NRP1-dependent. Indeed, combined inhibition of ß3 integrin and NRP1 decreased VEGF-mediated angiogenic responses further than individual inhibition of these receptors. We also show that avß3 integrin can associate with NRP1 in a VEGF-dependent fashion. Our data suggest that ß3 integrin may, in part, negatively regulate VEGF signaling by sequestering NRP1 and preventing it from interacting with VEGFR2

Endothelial FAK is required for tumour angiogenesis

Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that plays a fundamental role in integrin and growth factor mediated signalling and is an important player in cell migration and proliferation, processes vital for angiogenesis. However, the role of FAK in adult pathological angiogenesis is unknown. We have generated endothelial-specific tamoxifen-inducible FAK knockout mice by crossing FAK-floxed (FAKfl/fl) mice with the platelet derived growth factor b (Pdgfb)-iCreER mice. Tamoxifen-treatment of Pdgfb-iCreER;FAKfl/fl mice results in FAK deletion in adult endothelial cells (ECs) without any adverse effects. Importantly however, endothelial FAK-deletion in adult mice inhibited tumour growth and reduced tumour angiogenesis. Furthermore, in in vivo angiogenic assays FAK deletion impairs vascular endothelial growth factor (VEGF)-induced neovascularization. In addition, in vitro deletion of FAK in ECs resulted in reduced VEGF-stimulated Akt phosphorylation and correlating reduced cellular proliferation as well as increased cell death. Our data suggest that FAK is required for adult pathological angiogenesis and validates FAK as a possible target for anti-angiogenic therapies

Tumoural activation of TLR3–SLIT2 axis in endothelium drives metastasis

Blood vessels support tumours by providing nutrients and oxygen, while also acting as conduits for the dissemination of cancer1. Here we use mouse models of breast and lung cancer to investigate whether endothelial cells also have active 'instructive' roles in the dissemination of cancer. We purified genetically tagged endothelial ribosomes and their associated transcripts from highly and poorly metastatic tumours. Deep sequencing revealed that metastatic tumours induced expression of the axon-guidance gene Slit2 in endothelium, establishing differential expression between the endothelial (high Slit2 expression) and tumoural (low Slit2 expression) compartments. Endothelial-derived SLIT2 protein and its receptor ROBO1 promoted the migration of cancer cells towards endothelial cells and intravasation. Deleting endothelial Slit2 suppressed metastatic dissemination in mouse models of breast and lung cancer. Conversely, deletion of tumoural Slit2 enhanced metastatic progression. We identified double-stranded RNA derived from tumour cells as an upstream signal that induces expression of endothelial SLIT2 by acting on the RNA-sensing receptor TLR3. Accordingly, a set of endogenous retroviral element RNAs were upregulated in metastatic cells and detected extracellularly. Thus, cancer cells co-opt innate RNA sensing to induce a chemotactic signalling pathway in endothelium that drives intravasation and metastasis. These findings reveal that endothelial cells have a direct instructive role in driving metastatic dissemination, and demonstrate that a single gene (Slit2) can promote or suppress cancer progression depending on its cellular source

Haematopoietic focal adhesion kinase deficiency alters haematopoietic homeostasis to drive tumour metastasis

Metastasis is the main cause of cancer-related death and thus understanding the molecular and cellular mechanisms underlying this process is critical. Here, our data demonstrate, contrary to established dogma, that loss of haematopoietic-derived focal adhesion kinase (FAK) is sufficient to enhance tumour metastasis. Using both experimental and spontaneous metastasis models, we show that genetic ablation of haematopoietic FAK does not affect primary tumour growth but enhances the incidence of metastasis significantly. At a molecular level, haematopoietic FAK deletion results in an increase in PU-1 levels and decrease in GATA-1 levels causing a shift of hematopoietic homeostasis towards a myeloid commitment. The subsequent increase in circulating granulocyte number, with an increase in serum CXCL12 and granulocyte CXCR4 levels, was required for augmented metastasis in mice lacking haematopoietic FAK. Overall our findings provide a mechanism by which haematopoietic FAK controls cancer metastasis

FAK-heterozygous mice display enhanced tumour angiogenesis

Genetic ablation of endothelial focal adhesion kinase (FAK) can inhibit pathological angiogenesis, suggesting that loss of endothelial FAK is sufficient to reduce neovascularization. Here we show that reduced stromal FAK expression in FAK-heterozygous mice unexpectedly enhances both B16F0 and CMT19T tumour growth and angiogenesis. We further demonstrate that cell proliferation and microvessel sprouting, but not migration, are increased in serum-stimulated FAK-heterozygous endothelial cells. FAK-heterozygous endothelial cells display an imbalance in FAK phosphorylation at pY397 and pY861 without changes in Pyk2 or Erk1/2 activity. By contrast, serum-stimulated phosphorylation of Akt is enhanced in FAK-heterozygous endothelial cells and these cells are more sensitive to Akt inhibition. Additionally, low doses of a pharmacological FAK inhibitor, although too low to affect FAK autophosphorylation in vitro, can enhance angiogenesis ex vivo and tumour growth in vivo. Our results highlight a potential novel role for FAK as a nonlinear, dose-dependent regulator of angiogenesis where heterozygous levels of FAK enhance angiogenesis

Endothelial α3β1-Integrin Represses Pathological Angiogenesis and Sustains Endothelial-VEGF

Integrin a3ß1 is a major receptor for laminin. The expression levels of laminins-8 and -10 in the basement membrane surrounding blood vessels are known to change during tumor angiogenesis. Although some studies have suggested that certain ligands of a3ß1 can affect angiogenesis either positively or negatively, either a direct in vivo role for a3ß1 in this process or its mechanism of action in endothelial cells during angiogenesis is still unknown. Because the global genetic ablation of a3-integrin results in an early lethal phenotype, we have generated conditional-knockout mice where a3 is deleted specifically in endothelial cells (ec-a3-/-). Here we show that ec-a3-/- mice are viable, fertile, and display enhanced tumor growth, elevated tumor angiogenesis, augmented hypoxia-induced retinal angiogenesis, and increased vascular endothelial growth factor (VEGF)-mediated neovascularization ex vivo and in vivo. Furthermore, our data provide a novel method by which an integrin may regulate angiogenesis. We show that a3ß1 is a positive regulator of endothelial-VEGF and that, surprisingly, the VEGF produced by endothelial cells can actually repress VEGF-receptor 2 (Flk-1) expression. These data, therefore, identify directly that endothelial a3ß1 negatively regulates pathological angiogenesis and implicate an unexpected role for low levels of endothelial-VEGF as an activator of neovascularization