Purpose: Extensive work in preclinical models has shown that microenvironmental cells influence many aspects of cancer cell behavior, including metastatic potential and their sensitivity to... Show morePurpose: Extensive work in preclinical models has shown that microenvironmental cells influence many aspects of cancer cell behavior, including metastatic potential and their sensitivity to therapeutics. In the human setting, this behavior is mainly correlated with the presence of immune cells. Here, in addition to T cells, B cells, macrophages, and mast cells, we identified the relevance of nonimmune cell types for breast cancer survival and therapy benefit, including fibroblasts, myoepithelial cells, muscle cells, endothelial cells, and seven distinct epithelial cell types.Experimental Design: Using single-cell sequencing data, we generated reference profiles for all these cell types. We used these reference profiles in deconvolution algorithms to optimally detangle the cellular composition of more than 3,500 primary breast tumors of patients that were enrolled in the SCAN-B and MATADOR clinical trials, and for which bulk mRNA sequencing data were available.Results: This large data set enables us to identify and subsequently validate the cellular composition of miaoenvironments that distinguish differential survival and treatment benefit for different treatment regimens in patients with primary breast cancer. In addition to immune cells, we have identified that survival and therapy benefit are characterized by various contributions of distinct epithelial cell types.Conclusions: From our study, we conclude that differential survival and therapy benefit of patients with breast cancer are characterized by distinct microenvironments that include specific populations of immune and epithelial cells. Show less
Bot, I.; Velden, D. van der; Bouwman, M.; Kröner, M.J.; Kuiper, J.; Quax, P.H.A.; Vries, M.R. de 2020
Mast cells have been associated with arteriogenesis and collateral formation. In advancedhuman atherosclerotic plaques, mast cells have been shown to colocalize with plaque neovessels,and mast... Show moreMast cells have been associated with arteriogenesis and collateral formation. In advancedhuman atherosclerotic plaques, mast cells have been shown to colocalize with plaque neovessels,and mast cells have also been associated with tumor vascularization. Based on these associations,we hypothesize that mast cells promote angiogenesis during ischemia. In human ischemic muscletissue from patients with end-stage peripheral artery disease, we observed activated mast cells,predominantly located around capillaries. Also, in mouse ischemic muscles, mast cells were detectedduring the revascularization process and interestingly, mast cell activation status was enhanced up to10 days after ischemia induction. To determine whether mast cells contribute to both arteriogenesisand angiogenesis, mast cells were locally activated immediately upon hind limb ischemia in C57Bl/6mice. At day 9, we observed a 3-fold increase in activated mast cell numbers in the inguinal lymphnodes. This was accompanied by an increase in the amount of Ly6Chigh inflammatory monocytes.Interestingly, local mast cell activation increased blood flow through the hind limb (46% at day 9)compared to that in non-activated control mice. Histological analysis of the muscle tissue revealedthat mast cell activation did not aect the number of collaterals, but increased the collateral diameter,as well as the number of CD31+ capillaries. Together, these data illustrate that locally activated mastcell contribute to arteriogenesis and angiogenesis. Show less
Bot, I.; Velden, D. van der; Bouwman, M.; Kroner, M.J.; Kuiper, J.; Quax, P.H.A.; Vries, M.R. de 2020
Mast cells have been associated with arteriogenesis and collateral formation. In advanced human atherosclerotic plaques, mast cells have been shown to colocalize with plaque neovessels, and mast... Show moreMast cells have been associated with arteriogenesis and collateral formation. In advanced human atherosclerotic plaques, mast cells have been shown to colocalize with plaque neovessels, and mast cells have also been associated with tumor vascularization. Based on these associations, we hypothesize that mast cells promote angiogenesis during ischemia. In human ischemic muscle tissue from patients with end-stage peripheral artery disease, we observed activated mast cells, predominantly located around capillaries. Also, in mouse ischemic muscles, mast cells were detected during the revascularization process and interestingly, mast cell activation status was enhanced up to 10 days after ischemia induction. To determine whether mast cells contribute to both arteriogenesis and angiogenesis, mast cells were locally activated immediately upon hind limb ischemia in C57Bl/6 mice. At day 9, we observed a 3-fold increase in activated mast cell numbers in the inguinal lymph nodes. This was accompanied by an increase in the amount of Ly6C(high) inflammatory monocytes. Interestingly, local mast cell activation increased blood flow through the hind limb (46% at day 9) compared to that in non-activated control mice. Histological analysis of the muscle tissue revealed that mast cell activation did not affect the number of collaterals, but increased the collateral diameter, as well as the number of CD31(+) capillaries. Together, these data illustrate that locally activated mast cell contribute to arteriogenesis and angiogenesis. Show less
Lagraauw, H.M.; Wezel, A.; Velden, D. van der; Kuiper, J.; Bot, I. 2019
Mast cells accumulate in the perivascular tissue during atherosclerotic plaque progression and contribute to plaque destabilization. However, the specific triggers for mast cell activation in... Show moreMast cells accumulate in the perivascular tissue during atherosclerotic plaque progression and contribute to plaque destabilization. However, the specific triggers for mast cell activation in atherosclerosis remain unresolved. We hypothesized that psychological stress-induced activation of mast cells may contribute to plaque destabilization. To investigate this, apoE-/- mice on Western-type diet were exposed to 120' restraint stress. A single episode of restraint caused a significant increase in mast cell activation in the heart. In addition to a rise in serum corticosterone and changes in circulating leukocyte populations, we observed an increase in the circulating pro-inflammatory cytokine interleukin (IL)-6 in the stressed mice. Subsequent characterization of the atherosclerotic plaques revealed a high incidence and larger size of intraplaque hemorrhages in stressed mice. In mast cell-deficient apoE-/- mice, restraint stress affected circulating leukocyte levels, but did not increase plasma IL-6 levels. Furthermore, we did not observe any intraplaque hemorrhages in these mice upon stress, strongly indicating the involvement of a mast cell-dependent response to stress in atherosclerotic plaque destabilization. In conclusion, we demonstrate that acute stress activates mast cells, which induces the incidence of intraplaque hemorrhage in vivo, identifying acute stress as a risk factor for atherosclerotic plaque destabilization. Show less
Hermans, M.P.J.; Velden, D. van der; Cabezas, J.M.M.; Putter, H.; Huizinga, T.W.J.; Kuiper, J.; ... ; Woude, D. van der 2017
Background Patients with rheumatoid arthritis (RA) have a high risk of cardiovascular mortality, which is further increased in presence of anti–citrullinated protein antibodies (ACPA). Recently,... Show moreBackground Patients with rheumatoid arthritis (RA) have a high risk of cardiovascular mortality, which is further increased in presence of anti–citrullinated protein antibodies (ACPA). Recently, ACPA were unexpectedly detected in a cohort of patients with coronary artery disease (CAD) in the absence of RA. However, it is unknown if ACPA are consistently present in different cohorts of CAD patients, and if ACPA presence affects the course of cardiovascular disease in these patients. The purpose of this study was to assess the relationship between ACPA and long-term outcomes (including mortality) in patients with ST-elevation myocardial infarction (STEMI) in the absence of RA.Materials and methods Patients with from the MISSION! Intervention Study were included (n=275). The cohort consists of patients with STEMI, who subsequently underwent a percutaneous coronary intervention. Patients with RA were excluded. ACPA positivity was determined using a commercially available ELISA system (Quanta Lite CCP3.1 IgG/IgA). The association between ACPA (anti-CCP3) status at baseline and outcomes, such as re-infarction and death, was investigated during a 10 year follow-up.Results In total, 29 (11%) of 275 included patients were ACPA-positive, substantiating the previous description of ACPA in CAD patients. Increased cumulative cardiac mortality was observed in ACPA-positive patients in comparison with ACPA-negative patients. Moreover, after correction for additional associated factors (such as age, gender, body mass index, presence of diabetes, etc.), ACPA-positivity was still associated with increased long-term mortality (HR 3.1 [CI 1.4–7.1] p=0.01] and the long-term combined endpoint of re-infarction and death (HR 2.4 [1.2–4.6] p=0.01).Conclusions Among STEMI patients without RA, the presence of ACPA is independently associated with increased long-term mortality and the combined endpoint of re-infarction and death. We suggest that ACPA, previously solely linked to RA development and severity, may also play a role as an additional risk factor in cardiovascular disease. The hypothesis that ACPA might act as an independent pro-atherogenic factor in patients with and without RA, has to be investigated in subsequent studies. Show less
Hermans, M.P.J.; Volkov, M.; Velden, D. van der; Cabezas, J.M.M.; Huizinga, T.W.J.; Kuiper, J.; ... ; Woude, D. van der 2017
CC Chemokine Receptor 2 (CCR2) and its endogenous ligand CCL2 are involved in a number of diseases, including atherosclerosis. Several CCR2 antagonists have been developed as potential therapeutic... Show moreCC Chemokine Receptor 2 (CCR2) and its endogenous ligand CCL2 are involved in a number of diseases, including atherosclerosis. Several CCR2 antagonists have been developed as potential therapeutic agents, however their in vivo clinical efficacy was limited. In this report, we aimed to determine whether 15a, an antagonist with a long residence time on the human CCR2, is effective in inhibiting the development of atherosclerosis in a mouse disease model. First, radioligand binding assays were performed to determine affinity and binding kinetics of 15a on murine CCR2. To assess the in vivo efficacy, western-type diet fed apoE-/- mice were treated daily with 15a or vehicle as control. Treatment with 15a reduced the amount of circulating CCR2+ monocytes and the size of the atherosclerotic plaques in both the carotid artery and the aortic root. We then showed that the long pharmacokinetic half-life of 15a combined with the high drug concentrations ensured prolonged CCR2 occupancy. These data render 15a a promising compound for drug development and confirms high receptor occupancy as a key parameter when targeting chemokine receptors. Show less
Velden, D. van der; Lagraauw, H.M.; Wezel, A.; Launay, P.; Kuiper, J.; Huizinga, T.W.J.; ... ; Stoop, J.N. 2016
BACKGROUND\nRheumatoid arthritis (RA) is a multifactorial autoimmune disease, which is characterized by inflammation of synovial joints leading to the destruction of cartilage and bone.... Show moreBACKGROUND\nRheumatoid arthritis (RA) is a multifactorial autoimmune disease, which is characterized by inflammation of synovial joints leading to the destruction of cartilage and bone. Infiltrating mast cells can be found within the inflamed synovial tissue, however their role in disease pathogenesis is unclear. Therefore we have studied the role of mast cells during different phases of experimental arthritis.\nMETHODS\nWe induced collagen-induced arthritis (CIA), the most frequently used animal model of arthritis, in an inducible mast cell knock-out mouse and determined the effect of mast cell depletion on the development and severity of arthritis.\nRESULTS\nDepletion of mast cells in established arthritis did not affect clinical outcome. However, depletion of mast cells during the preclinical phase resulted in a significant reduction in arthritis. This reduction coincided with a decrease in circulating CD4(+) T cells and inflammatory monocytes but not in the collagen-specific antibody levels. Mast cell depletion resulted in reduced levels of IL-6 and IL-17 in serum. Furthermore, stimulation of splenocytes from mast cell-depleted mice with collagen type II resulted in reduced levels of IL-17 and enhanced production of IL-10.\nCONCLUSIONS\nHere we show that mast cells contribute to the preclinical phase of CIA. Depletion of mast cells before disease onset resulted in an altered collagen-specific T cell and cytokine response. These data may suggest that mast cells play a role in the regulation of the adaptive immune response during the development of arthritis. Show less
In dit proefschrift hebben wij de invloed van mestcellen in dierexperimentele modellen voor reumatoïde artritis en aderverkalking bestudeerd met een focus op de rol van mestcellen in de (sub)... Show moreIn dit proefschrift hebben wij de invloed van mestcellen in dierexperimentele modellen voor reumatoïde artritis en aderverkalking bestudeerd met een focus op de rol van mestcellen in de (sub)-klinische fases van het ziekteproces, waarin er een actieve immuunrespons ontwikkeld was. Ook hebben wij de aanwezigheid van verschillende soorten antistoffen in het serum van cardiovasculaire patiënten onderzocht. Wij hebben de niveaus van antilichamen gecorreleerd aan klinische observaties zoals body mass index (BMI), lipidenprofiel, klinische diagnose van vaatlijden, samenstelling van de atherosclerotische plaque en de uitkomst van de ziekte. Show less
Woude, D. van der; Hermans, M.P.; Velden, D. van der; Trouw, L.A.; Huizinga, T.W.; Kuiper, J.; ... ; Toes, R.E. 2016
Background Anti-citrullinated protein antibodies (ACPA) are thought to be highly specific for RA. ACPA are associated with risk factors for RA and with joint destruction, and are therefore presumed... Show moreBackground Anti-citrullinated protein antibodies (ACPA) are thought to be highly specific for RA. ACPA are associated with risk factors for RA and with joint destruction, and are therefore presumed to be involved in RA pathogenesis. ACPA-positive RA patients also have increased cardiovascular mortality (1). In cardiovascular disease, inflammatory changes occur in vessel walls, raising the question whether ACPA (presumed to be pro-inflammatory) may contribute to this process.Objectives To investigate the prevalence and prognostic implications of ACPA in patients with cardiovascular disease without RA.Methods ACPA were determined by anti-CCP3 commercial assay in baseline sera from 290 patients with ST-elevation myocardial infarction participating in the MISSION intervention study (2). Patients with RA were excluded. The association between ACPA and long-term mortality was investigated.Results 30/290 (10.3%) of the non-RA patients with cardiovascular disease were ACPA-positive. ACPA-positive non-RA patients had a significantly increased long-term cardiac mortality compared to ACPA-negative non-RA patients (Figure). Corrected for age, ACPA positivity was independently associated with long-term mortality [HR 2.4 (CI 1.1–5.4) p-Value=0.026].Conclusions ACPA can be detected in a considerable proportion of non-RA patients with cardiovascular disease. This challenges the presumed specificity of ACPA for RA. In both RA and cardiovascular disease, ACPA are associated with a worse disease outcome possibly by an ACPA-specific enhancement of inflammation. Future studies into ACPA in patients with cardiovascular disease offer the opportunity to dissect which risk factors are associated with ACPA in RA versus non-RA patients. This may supply crucial insights into the development of this autoimmune reaction. Show less
Velden, D. van der; Lagraauw, H.M.; Wezel, A.; Launay, P.; Kuiper, J.; Huizinga, T.W.J.; ... ; Stoop, J.N. 2016
Mast cells accumulate in the perivascular tissue during plaque progression and upon activation release pro-inflammatory cytokines and extracellular matrix degrading enzymes which fuel the... Show moreMast cells accumulate in the perivascular tissue during plaque progression and upon activation release pro-inflammatory cytokines and extracellular matrix degrading enzymes which fuel the inflammatory process and decrease lesion stability. Interestingly, mast cells have been shown to co-localize with peripheral neurons and contain receptors for various hormones and neuropeptides which are released upon exposure to stress. Therefore, we hypothesize that stress-induced neuronal activation of mast cells contributes to plaque destabilization.In apoE−/− mice the acute stress induced by 120′ restraint caused a significant increase in mast cell activation in the heart (37.3 ± 1.8% vs. 50.1 ± 4.9%). In parallel with a rise in serum corticosterone levels we observed a transient increase mast cell-specific β-hexosaminidase levels and the pro-inflammatory cytokine, interleukin-6 in the stressed mice compared to controls (55.9 ± 11.0 pg/ml vs. 29.6 ± 5.49 pg/ml). Subsequent characterization of atherosclerotic lesions in the aortic root revealed a significant reduction in lesion collagen content and a higher incidence of intraplaque hemorrhages, a hallmark of the vulnerable plaque (38.5% vs. 7.7%).Importantly, both these effects were reduced in mice treated with the mast cell stabilizer cromolyn and completely abolished in mast cell deficient (apoE−/−/Kit(W-sh/W-sh)) mice, thus strongly indicating the involvement of a mast cell-dependent response to stress in atherosclerotic plaque destabilization.We demonstrate that acute stress activates mast cells near atherosclerotic lesions and contributes to plaque destabilization, identifying acute stress as a risk factor for acute cardiovascular syndromes. Show less
Hermans, M.; Velden, D. van der; Huizinga, T.W.J.; Kuiper, J.; Toes, R.E.M.; Schalij, M.J.; ... ; Woude, D. van der 2016
Background Rheumatoid arthritis (RA) is a multifactorial autoimmune disease, which is characterized by inflammation of synovial joints leading to the destruction of cartilage and bone. Infiltrating... Show moreBackground Rheumatoid arthritis (RA) is a multifactorial autoimmune disease, which is characterized by inflammation of synovial joints leading to the destruction of cartilage and bone. Infiltrating mast cells can be found within the inflamed synovial tissue, however their role in disease pathogenesis is unclear. Therefore we have studied the role of mast cells during different phases of experimental arthritis. Methods We induced collagen-induced arthritis (CIA), the most frequently used animal model of arthritis, in an inducible mast cell knock-out mouse and determined the effect of mast cell depletion on the development and severity of arthritis. Results Depletion of mast cells in established arthritis did not affect clinical outcome. However, depletion of mast cells during the preclinical phase resulted in a significant reduction in arthritis. This reduction coincided with a decrease in circulating CD4+ T cells and inflammatory monocytes but not in the collagen-specific antibody levels. Mast cell depletion resulted in reduced levels of IL-6 and IL-17 in serum. Furthermore, stimulation of splenocytes from mast cell-depleted mice with collagen type II resulted in reduced levels of IL-17 and enhanced production of IL-10. Conclusions Here we show that mast cells contribute to the preclinical phase of CIA. Depletion of mast cells before disease onset resulted in an altered collagen-specific T cell and cytokine response. These data may suggest that mast cells play a role in the regulation of the adaptive immune response during the development of arthritis. Keywords Mast cells Collagen-induced arthritis RMB mice T cells Show less
