ReviewThe reciprocal function and regulation of tumor vessels and immune cells offers new therapeutic opportunities in cancer
Introduction
The onset of tumor neovascularization is a well-established hallmark of cancer that is initiated at a certain time during tumor progression depending on the tumor type, grade and stage. Blood vessels not only have to provide oxygen and deplete waste products to oblige the demands of a growing tumor but also form niches that enable cancer stem cell maintenance, protection and entail sites of infiltrating immune cells [[1], [2], [3], [4]]. Various mechanisms for the reinstatement of blood vessel growth have been described of which angiogenesis, the sprouting of new vessels from pre-existing capillaries and postcapillary venules is the most commonly used mode [[5], [6], [7]]. Subsequently, tip cells anastomose with tip cells from neighboring sprouts to connect the newly formed vascular structures with the help of bridging macrophages. With the initiation of blood flow, the establishment of a basement membrane, and the recruitment of vessel-stabilizing pericytes, vascular growth is then terminated [5,6]. Thus, under physiological conditions angiogenesis is tightly regulated by a homeostatic balance of a wealth of proangiogenic and inhibitory factors of which members of the vascular endothelial growth factor (VEGF), and angiopoietin tyrosine kinase receptor families as well as various members of the CXC and CC chemokines are the most prominent molecules to orchestrate this multistep process [1,8]. The balance of angiogenesis-promoting and inhibiting molecules is, however, lost in tumors. Upon initiating neovascularization, tumors continue to promote angiogenic signaling for several reasons [9]. First, tumors express oncogenes and/or lose tumor suppressor genes that result in the activation of pro-angiogenic signaling pathways [[10], [11], [12]]. Second, expanding tumors exhibit a continually and abnormally growing tumor vasculature with a typical display of irregular-shaped, hyperdilated and poorly pericyte-covered tumor vessels that cause a leaky and sluggish blood flow [13]. These typical signs of poor vessel maturation increase interstitial pressure and generate and maintain a hypoxic and acidic environment that continues to elevate proangiogenic factors thereby further aggravating a pro-angiogenic feedback loop that never ceases.
Effects of low-oxygen tension are mediated by hypoxia-inducible factor (HIF) transcription factors that coordinate a transcriptional program to ensure metabolic and vascular adaptations under hypoxic conditions by various mechanisms [[14], [15], [16]]. Hypoxia upregulates various proangiogenic growth factors and chemokines that directly engage in vessel growth like VEGF, PlGF and Ang2 [[17], [18], [19], [20], [21]]. In addition, numerous experimental models have shown that both hypoxic and acidic conditions in the tumor microenvironment change the availability of metabolites and induce the secretion of molecules in tumors (e.g. CSF1, GM-CSF, G-CSF, CX3CL1, CXCL5, CXCL12, CCL17, CCL22, IL6, Sema3) that attract innate immune cells to the tumor site where they become reprogrammed to serve as an additional source of angiogenic factors [[22], [23], [24], [25], [26], [27], [28]]. It is important to note that innate immune cells elicit a high plasticity in order to quickly adapt and serve the homeostatic repair program. Upon injury, first neutrophils and then macrophages are recruited to the wound that are immunosupportive and angiostatic due to their responsibility to phagocytose and kill bacteria and degrade necrotic tissue. In the subsequent resolution phase, myeloid cells then convert to an angiogenic and immunosuppressive phenotype to support tissue and blood vessel restoration [29]. The ability of myeloid cells to display such opposing properties to either eliminate potential infections or hinder excessive inflammation is indeed pivotal to properly regulate tissue injury and maintain tissue homeostasis [30]. In line with the concept that tumors are wounds that never heal [31], tumors take advantage of the myeloid plasticity by reprogramming these cells to exhibit both immunosuppressive and angiogenic features to enable tumors to escape immunosurveillance and facilitate vascular expansion. These two features have been defined as pivotal hallmarks in cancer propagation and progression [32].
There is increasing evidence that tumor-supporting inflammation and angiogenesis, although being distinct and separable processes, are closely related events that are in part regulated by common chemokines [32,33]. VEGF and Ang2 are proangiogenic factors that directly and indirectly convey immunosuppression in endothelial cells as well as innate and adaptive immune cells [[34], [35], [36], [37]]. VEGF induces an immunosuppressive vasculature on different levels (Fig.1). It downregulates the expression of the vascular adhesion molecules I-CAM1 and V-CAM1 that are necessary for T-cells to cross the endothelial layer and transit into the tumor. VEGF and Ang2 have been shown to enhance the expression of the negative checkpoint regulator programmed cell death protein 1 (PD-1)–programmed cell death 1 ligand 1 (PD-L1) in endothelial cells thus disabling the cytotoxic function of PD1+-T cells [[38], [39], [40], [41]]. In addition, elevated FasL levels on tumor ECs is known to trigger apoptosis of Fas-expressing CD8 + T cells [42]. Tumor-associated ECs also preferentially attract immunosuppressive Tregs by upregulating the multifunctional endothelial receptor CLEVER-1/stabilin-1 [43]. All these mechanisms consequently lead to a potent barrier that disables cytotoxic T cell infiltration into the tumor. VEGF, however, not only facilitates VEGFR-2 signaling in endothelial cells but also in VEGFR2+ immune cells. Thereby, it stimulates the expression of PD-L1 in dendritic cells and blocks the maturation and thus, functionality of dendritic cells due to their impaired ability to present tumor antigens [44]. VEGF/VEGFR2 signaling further suppresses proliferation of effector T cells but increases and promotes tumor homing of Tregs [32,44]. This effect is dependent on neuropilin-1 (nrp1) because in an experimental tumor model, nrp1 depletion resulted in reduction of Tregs with a concomitant increase in cytotoxic T cells [45]. VEGF also upregulates several negative immune checkpoint receptors besides PD-1 (e.g. CTLA-4, Lag-3, TIM-3) on T cells contributing to T-cell anergy and exhaustion [46]. Finally, elevated levels of VEGF and/or Ang-2 as well as other tumor-secreted factors promote the recruitment and expansion of innate immune cell populations including TAMs, TEMs, neutrophils, MDSC and immature dendritic cells that secrete molecules fostering an immunosuppressive and angiogenic tumor microenvironment.
Section snippets
Innate immune cells promote angiogenesis
As discussed above, tumor-associated macrophages (TAM) and tumor-associated neutrophils (TAN) can either convey angiostatic and immunosupportive or proangiogenic and immunosuppressive features, but in tumors are commonly found to polarize to an immunosuppressive as well as angiogenic phenotype [[47], [48], [49]]. In addition, immature Gr1+ cells with either a mononuclear or granular morphology have been identified in tumors that promote immunosuppressive functions and are therefore also termed
Metabolic pathways in immune cells regulate angiogenesis
The tumor induces a major disturbance in tissue homeostasis and cellular metabolism by endorsing a hypoxic and acidic microenvironment that strongly affects metabolic availability not only for tumor cells but also other cell constituents within the tumor (Fig.2). Although it is well-established that intratumoral hypoxia induces the recruitment of immunosuppressive and angiogenic myeloid cells to the tumor site, it is less understood how subsequent changes in metabolite availability for immune
Immune cells facilitate resistance to antiangiogenic therapy
Compelling data from multiple laboratories have provided a convincing rationale for the development of VEGF and VEGF receptor inhibitors to block intratumoral vascular growth and subsequent tumor propagation. Subsequently, bevacizumab (Avastin, Genentech/Roche), a monoclonal antibody against VEGF, as well as sorafenib (Nexavar, Bayer) and sunitinib (Sutent, Pfizer), both kinase inhibitors that target VEGF receptor (VEGFR) were the first FDA-approved angiogenic inhibitors followed by other
Antiangiogenic therapy meets immunotherapy
Various studies including those mentioned above, have provided compelling evidence that antiangiogenic therapy is most efficacious when an immunostimulatory environment is generated. This is in agreement with the concept of vascular normalization to reduce hypoxia and polarize innate immune cells to an immunosupportive phenotype. Emerging from these studies is the proposition that angiogenesis and inflammation are reciprocally regulated and that immune cells play a pivotal role in regulating
Conclusions
Currently, various combinatorial treatment modalities of angiogenic inhibitors, specifically those targeting the VEGF/R and Ang2/Tie axis with immune checkpoint inhibitors and other immunotherapies are evaluated in the clinical setting [116]. These trials have been initiated based on compelling evidence from mouse tumor model systems that such a combination modulates both the tumor vasculature and the immune system to foster an immunosupportive environment and thus improves and endures efficacy
Sources of funding
This work was supported by grants from the ERC CoG ImmunoFIT Methusalem (to MM) and from the Flamish government FWO (G066515N) (to MM) and FWO (G0A0818N) (to GB) and the Belgian Association Against Cancer for the project 2014-197 (to MM) and from the National Institute of Health NIH/NCI (to GB).
Competing interests
None.
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