Helene Rundqvist - Academia.edu (original) (raw)
Papers by Helene Rundqvist
Frontiers in Immunology
Myeloid cell interactions with cells of the adaptive immune system are an essential aspect of imm... more Myeloid cell interactions with cells of the adaptive immune system are an essential aspect of immunity. A key aspect of that interrelationship is its modulation by the microenvironment. Oxygen is known to influence myelosuppression of T cell activation in part via the Hypoxia inducible (HIF) transcription factors. A number of drugs that act on the HIF pathway are currently in clinical use and it is important to evaluate how they act on immune cell function as part of a better understanding of how they will influence patient outcomes. We show here that increased activation of the HIF pathway, either through deletion of the negative regulator of HIF, the von Hippel-Lindau (VHL) gene, in myeloid cells, or through pharmacological inhibitors of VHL-mediated degradation of HIF, potently suppresses T cell proliferation in myeloid cell/T cell culture. These data demonstrate that both pharmacological and genetic activation of HIF in myeloid cells can suppress adaptive cell immune response.
Arteriosclerosis, Thrombosis, and Vascular Biology, 2015
Introduction: Mechanisms that regulate the positive association between thrombosis and metastasis... more Introduction: Mechanisms that regulate the positive association between thrombosis and metastasis are incompletely understood. It was hypothesised that thrombus formation stimulates a hypoxic response, which in turn promotes extravasation. The primary aim was to determine whether thrombosis of the pulmonary microvasculature (T pm ) increases extravasation via myeloid (neutrophil and macrophage) hypoxia-inducible factor (HIF). Methods and Results: T pm was induced in wild type mice via tail vein administration of polystyrene microbeads (15μm diameter, 1000/mouse). T pm led to chronological increases in pulmonary HIF1α expression (P=0.01), HIF2α expression (P<0.01), neutrophil infiltration (P<0.05), and macrophage infiltration (P<0.05; 1-5days post-T pm vs. non-thrombosed vehicle controls, n=8/group/time point); these increases were comparable with changes observed following vena cava thrombosis (assessed via image analysis of immunostained tissue throughout). In wild type mi...
fibroblasts with cancer cells in vitro: gene network analysis of view on the process and details ... more fibroblasts with cancer cells in vitro: gene network analysis of view on the process and details at the gene/protein level. The combination of our methods pointed to proteins, such as members of the Rho pathway, pro-inflammatory signature and the YAP1/TAZ cascade, that warrant further fibroblasts
CD8+ T cells infiltrate virtually every tissue to find and destroy infected or mutated cells. The... more CD8+ T cells infiltrate virtually every tissue to find and destroy infected or mutated cells. They often traverse varying oxygen levels and nutrient-deprived microenvironments. High glycolytic activity in tissues can result in extended exposure of cytotoxic T cells to the metabolite lactate. Lactate can be immunosuppressive, at least in part due to its association with tissue acidosis. We show here that the lactate anion is well tolerated by CD8+ T cells in pH neutral conditions. We describe how lactate is taken up by activated CD8+ T cells and is capable of displacing glucose as a carbon source. Activation in the presence of a pH neutral form of lactate significantly alters the CD8+ T cell transcriptome, including the expression of key effector differentiation markers such as granzyme B and interferon-gamma. Our studies reveal the novel metabolic features of lactate utilization by activated CD8+ T cells, and highlight the importance of lactate in shaping the differentiation and act...
Animals require an immediate response to oxygen availability to allow rapid shifts between oxidat... more Animals require an immediate response to oxygen availability to allow rapid shifts between oxidative and glycolytic metabolism. These metabolic shifts are highly regulated by the HIF transcription factor. The Factor Inhibiting HIF (FIH) is an asparaginyl hydroxylase that controls HIF transcriptional activity in an oxygen-dependent manner. We show here that FIH loss increases oxidative metabolism, while also increasing glycolytic capacity, and that this gives rise to an increase in oxygen consumption. We further show that the loss of FIH acts to accelerate the cellular metabolic response to hypoxia. Skeletal muscle expresses 50-fold higher levels of FIH than other tissues: we analyzed skeletal muscle FIH mutants, and found a decreased metabolic efficiency, correlated with an increased oxidative rate and an increased rate of hypoxic response. We find that FIH, through its regulation of oxidation, acts in concert with the PHD/VHL pathway to accelerate HIF-mediated metabolic responses to hypoxia.
Circulation Research, 2021
Rationale: Ischemic injuries remain a leading cause of mortality and morbidity worldwide, and res... more Rationale: Ischemic injuries remain a leading cause of mortality and morbidity worldwide, and restoration of functional blood perfusion is vital to limit tissue damage and support healing. Objective: To reveal a novel role of macrophages in reestablishment of functional tissue perfusion following ischemic injury that can be targeted to improve tissue restoration. Methods and Results: Using intravital microscopy of ischemic hindlimb muscle in mice, and confocal microscopy of human tissues from amputated legs, we found that macrophages accumulated perivascularly in ischemic muscles, where they expressed high levels of iNOS (inducible nitric oxide [NO] synthase). Genetic depletion of iNOS specifically in macrophages (Cx3cr1-CreERT2;Nos2 fl/fl or LysM-Cre;Nos2 fl/fl ) did not affect vascular architecture but highly compromised blood flow regulation in ischemic but not healthy muscle, which resulted in aggravated ischemic damage. Thus, the ability to upregulate blood flow was shifted fro...
Cell Death Discovery, 2020
Cancer-associated fibroblasts (CAFs) promote tumor growth and progression, and increase drug resi... more Cancer-associated fibroblasts (CAFs) promote tumor growth and progression, and increase drug resistance through several mechanisms. We have investigated the effect of CAFs on the p53 response to doxorubicin in prostate cancer cells. We show that CAFs produce interleukin-6 (IL-6), and that IL-6 attenuates p53 induction and upregulation of the pro-apoptotic p53 target Bax upon treatment with doxorubicin. This is associated with increased levels of MDM2 mRNA, Mdm2 protein bound to p53, and ubiquitinated p53. IL-6 also inhibited doxorubicin-induced cell death. Inhibition of JAK or STAT3 alleviated this effect, indicating that IL-6 attenuates p53 via the JAK/STAT signaling pathway. These results suggest that CAF-derived IL-6 plays an important role in protecting cancer cells from chemotherapy and that inhibition of IL-6 could have significant therapeutic value.
eLife, 2020
Exercise has a wide range of systemic effects. In animal models, repeated exertion reduces malign... more Exercise has a wide range of systemic effects. In animal models, repeated exertion reduces malignant tumor progression, and clinically, exercise can improve outcome for cancer patients. The etiology of the effects of exercise on tumor progression are unclear, as are the cellular actors involved. We show here that in mice, exercise-induced reduction in tumor growth is dependent on CD8+ T cells, and that metabolites produced in skeletal muscle and excreted into plasma at high levels during exertion in both mice and humans enhance the effector profile of CD8+ T-cells. We found that activated murine CD8+ T cells alter their central carbon metabolism in response to exertion in vivo, and that immune cells from trained mice are more potent antitumor effector cells when transferred into tumor-bearing untrained animals. These data demonstrate that CD8+ T cells are metabolically altered by exercise in a manner that acts to improve their antitumoral efficacy.
Cardio-Oncology, 2021
Background Adjuvant systemic breast cancer treatment improves disease specific outcomes, but also... more Background Adjuvant systemic breast cancer treatment improves disease specific outcomes, but also presents with cardiac toxicity. In this post-hoc exploratory analysis of the OptiTrain trial, the effects of exercise on cardiotoxicity were monitored by assessing fitness and biomarkers over the intervention and into survivorship. Methods; Women starting chemotherapy were randomized to 16-weeks of resistance and high-intensity interval training (RT-HIIT), moderate-intensity aerobic and high-intensity interval training (AT–HIIT), or usual care (UC). Outcome measures included plasma troponin-T (cTnT), Nt-pro-BNP and peak oxygen uptake (VO2peak), assessed at baseline, post-intervention, and at 1- and 2-years. Results For this per-protocol analysis, 88 women met criteria for inclusion. Plasma cTnT increased in all groups post-intervention. At the 1-year follow-up, Nt-pro-BNP was lower in the exercise groups compared to UC. At 2-years there was a drop in VO2peak for patients with high cTnT ...
Medicine & Science in Sports & Exercise, 2020
The FASEB Journal, 2007
11 subjects performed one-legged exercise program for 5 weeks. The subjects exercised one leg for... more 11 subjects performed one-legged exercise program for 5 weeks. The subjects exercised one leg for 45 min with restricted blood flow, followed by exercise with the other leg at the same absolute workload with unrestricted blood flow. Gene expression of MMP-2, -9, -14 and TIMP-1 were measured in m vastus lateralis before and after the training period, as well as 2 h after the first and last exercise bouts. MMP-14 and MMP-2 both increased robustly with training regardless of training condition, but with no increase after a single exercise bout. MMP-9 increased transiently after a single bout of exercise in the restricted leg, but no increase was evident with training. TIMP-1 had a tendency to increase after a single bout of exercise in the restricted leg and increased with training in both conditions, however more in the restricted leg. This support the importance of MMP for the adaptation and remodelling of the skeletal muscle with exercise training. The difference in expression of MMP-9 and TIMP-1 between ...
Adoptive transfer of anti-tumor cytotoxic T cells is a novel form of cancer immunotherapy, and a ... more Adoptive transfer of anti-tumor cytotoxic T cells is a novel form of cancer immunotherapy, and a key challenge is to ensure the survival and function of the transferred T cells. Immune cell survival requires adaptation to different micro-environments, and particularly to the hypoxic milieu of solid tumors. The HIF transcription factors are an essential aspect of this adaptation, and we undertook experiments to define structural determinants of HIF that would potentiate anti-tumor efficacy in cytotoxic T cells. We created retroviral vectors to deliver ectopic expression of HIF-1ɑ and HIF-2ɑ in mouse CD8+ T cells, together or individually, and with or without sensitivity to their oxygen-dependent inhibitors Von Hippel-Lindau (VHL) and Factor Inhibiting HIF (FIH). We found that HIF-2ɑ, but not HIF-1ɑ, drives broad transcriptional changes in CD8+ T cells, resulting in increased cytotoxic differentiation and cytolytic function against tumor targets. We further found that a specific mutat...
Medicine & Science in Sports & Exercise, 2020
PURPOSE:Regular physical exercise provides a significant risk reduction for breast cancer and rec... more PURPOSE:Regular physical exercise provides a significant risk reduction for breast cancer and recent studies suggest beneficial effects also on disease specific recurrence and mortality. However, little is known about how exercise exerts its protective effects. The primary aim of this study to evaluate the effect of voluntary running on tumor progression and metastasis in the PyMT mouse model of breast cancer. METHODS:From 4 weeks of age, female MMTV-PyMT mice on the FVB background were housed with access either to wirelessly recording running wheels or locked control wheels. Tumor growth was monitored continuously, tumor stage and pulmonary metastases were determined histologically at the 12 week endpoint. In a follow up study, pre-trained female FVB mice were injected intravenously with 2*10PyMT derived tumor cells (IC3) and after an additional 10 weeks of voluntary running, pulmonary metastases and immune cell infiltration was quantified (histologically and with flow cytometry). The CD8+ T-cell population was deleted using weekly administration of CD8 specific antibodies. RESULTS: PyMT mice average running distance was 6.4±2.4 km/day. No significant effects of voluntary running on tumorinitiation, volume or stage were found. However, a reduced number of metastases were observed in mice with access to running wheels (Ctrl 22.0±6.8 and Runners 9.1±1.7). Significant reductions in pulmonary metastasis frequency were also found in runners after intravenous injections of tumor cells (Ctrl 5.2±1.1 and Runners 1.9±0.7) and runningmice had a lower number ofmetastases with a high proliferation score. Metastatic lesions from running mice showed higher content of Granzyme B positive cells (Ctrl 1.2±0.5 and Runners 4.9±1.1), indicating an increased infiltration of cytotoxic T-cells. Depletion of CD8+ cells abolished the reduction in metastatic burden found in running mice compared to non-running mice All data is presented as Mean±SEM. CONCLUSIONS: In this highly aggressive, genetic, breast cancer model, an average of 6 km/day of voluntary running showed little effect on tumor formation and growth. However, the findings suggest that physical activity reduced outgrowth of metastatic lesions through an increased infiltration of cytotoxic immune cells.
Frontiers in Immunology
Myeloid cell interactions with cells of the adaptive immune system are an essential aspect of imm... more Myeloid cell interactions with cells of the adaptive immune system are an essential aspect of immunity. A key aspect of that interrelationship is its modulation by the microenvironment. Oxygen is known to influence myelosuppression of T cell activation in part via the Hypoxia inducible (HIF) transcription factors. A number of drugs that act on the HIF pathway are currently in clinical use and it is important to evaluate how they act on immune cell function as part of a better understanding of how they will influence patient outcomes. We show here that increased activation of the HIF pathway, either through deletion of the negative regulator of HIF, the von Hippel-Lindau (VHL) gene, in myeloid cells, or through pharmacological inhibitors of VHL-mediated degradation of HIF, potently suppresses T cell proliferation in myeloid cell/T cell culture. These data demonstrate that both pharmacological and genetic activation of HIF in myeloid cells can suppress adaptive cell immune response.
Arteriosclerosis, Thrombosis, and Vascular Biology, 2015
Introduction: Mechanisms that regulate the positive association between thrombosis and metastasis... more Introduction: Mechanisms that regulate the positive association between thrombosis and metastasis are incompletely understood. It was hypothesised that thrombus formation stimulates a hypoxic response, which in turn promotes extravasation. The primary aim was to determine whether thrombosis of the pulmonary microvasculature (T pm ) increases extravasation via myeloid (neutrophil and macrophage) hypoxia-inducible factor (HIF). Methods and Results: T pm was induced in wild type mice via tail vein administration of polystyrene microbeads (15μm diameter, 1000/mouse). T pm led to chronological increases in pulmonary HIF1α expression (P=0.01), HIF2α expression (P<0.01), neutrophil infiltration (P<0.05), and macrophage infiltration (P<0.05; 1-5days post-T pm vs. non-thrombosed vehicle controls, n=8/group/time point); these increases were comparable with changes observed following vena cava thrombosis (assessed via image analysis of immunostained tissue throughout). In wild type mi...
fibroblasts with cancer cells in vitro: gene network analysis of view on the process and details ... more fibroblasts with cancer cells in vitro: gene network analysis of view on the process and details at the gene/protein level. The combination of our methods pointed to proteins, such as members of the Rho pathway, pro-inflammatory signature and the YAP1/TAZ cascade, that warrant further fibroblasts
CD8+ T cells infiltrate virtually every tissue to find and destroy infected or mutated cells. The... more CD8+ T cells infiltrate virtually every tissue to find and destroy infected or mutated cells. They often traverse varying oxygen levels and nutrient-deprived microenvironments. High glycolytic activity in tissues can result in extended exposure of cytotoxic T cells to the metabolite lactate. Lactate can be immunosuppressive, at least in part due to its association with tissue acidosis. We show here that the lactate anion is well tolerated by CD8+ T cells in pH neutral conditions. We describe how lactate is taken up by activated CD8+ T cells and is capable of displacing glucose as a carbon source. Activation in the presence of a pH neutral form of lactate significantly alters the CD8+ T cell transcriptome, including the expression of key effector differentiation markers such as granzyme B and interferon-gamma. Our studies reveal the novel metabolic features of lactate utilization by activated CD8+ T cells, and highlight the importance of lactate in shaping the differentiation and act...
Animals require an immediate response to oxygen availability to allow rapid shifts between oxidat... more Animals require an immediate response to oxygen availability to allow rapid shifts between oxidative and glycolytic metabolism. These metabolic shifts are highly regulated by the HIF transcription factor. The Factor Inhibiting HIF (FIH) is an asparaginyl hydroxylase that controls HIF transcriptional activity in an oxygen-dependent manner. We show here that FIH loss increases oxidative metabolism, while also increasing glycolytic capacity, and that this gives rise to an increase in oxygen consumption. We further show that the loss of FIH acts to accelerate the cellular metabolic response to hypoxia. Skeletal muscle expresses 50-fold higher levels of FIH than other tissues: we analyzed skeletal muscle FIH mutants, and found a decreased metabolic efficiency, correlated with an increased oxidative rate and an increased rate of hypoxic response. We find that FIH, through its regulation of oxidation, acts in concert with the PHD/VHL pathway to accelerate HIF-mediated metabolic responses to hypoxia.
Circulation Research, 2021
Rationale: Ischemic injuries remain a leading cause of mortality and morbidity worldwide, and res... more Rationale: Ischemic injuries remain a leading cause of mortality and morbidity worldwide, and restoration of functional blood perfusion is vital to limit tissue damage and support healing. Objective: To reveal a novel role of macrophages in reestablishment of functional tissue perfusion following ischemic injury that can be targeted to improve tissue restoration. Methods and Results: Using intravital microscopy of ischemic hindlimb muscle in mice, and confocal microscopy of human tissues from amputated legs, we found that macrophages accumulated perivascularly in ischemic muscles, where they expressed high levels of iNOS (inducible nitric oxide [NO] synthase). Genetic depletion of iNOS specifically in macrophages (Cx3cr1-CreERT2;Nos2 fl/fl or LysM-Cre;Nos2 fl/fl ) did not affect vascular architecture but highly compromised blood flow regulation in ischemic but not healthy muscle, which resulted in aggravated ischemic damage. Thus, the ability to upregulate blood flow was shifted fro...
Cell Death Discovery, 2020
Cancer-associated fibroblasts (CAFs) promote tumor growth and progression, and increase drug resi... more Cancer-associated fibroblasts (CAFs) promote tumor growth and progression, and increase drug resistance through several mechanisms. We have investigated the effect of CAFs on the p53 response to doxorubicin in prostate cancer cells. We show that CAFs produce interleukin-6 (IL-6), and that IL-6 attenuates p53 induction and upregulation of the pro-apoptotic p53 target Bax upon treatment with doxorubicin. This is associated with increased levels of MDM2 mRNA, Mdm2 protein bound to p53, and ubiquitinated p53. IL-6 also inhibited doxorubicin-induced cell death. Inhibition of JAK or STAT3 alleviated this effect, indicating that IL-6 attenuates p53 via the JAK/STAT signaling pathway. These results suggest that CAF-derived IL-6 plays an important role in protecting cancer cells from chemotherapy and that inhibition of IL-6 could have significant therapeutic value.
eLife, 2020
Exercise has a wide range of systemic effects. In animal models, repeated exertion reduces malign... more Exercise has a wide range of systemic effects. In animal models, repeated exertion reduces malignant tumor progression, and clinically, exercise can improve outcome for cancer patients. The etiology of the effects of exercise on tumor progression are unclear, as are the cellular actors involved. We show here that in mice, exercise-induced reduction in tumor growth is dependent on CD8+ T cells, and that metabolites produced in skeletal muscle and excreted into plasma at high levels during exertion in both mice and humans enhance the effector profile of CD8+ T-cells. We found that activated murine CD8+ T cells alter their central carbon metabolism in response to exertion in vivo, and that immune cells from trained mice are more potent antitumor effector cells when transferred into tumor-bearing untrained animals. These data demonstrate that CD8+ T cells are metabolically altered by exercise in a manner that acts to improve their antitumoral efficacy.
Cardio-Oncology, 2021
Background Adjuvant systemic breast cancer treatment improves disease specific outcomes, but also... more Background Adjuvant systemic breast cancer treatment improves disease specific outcomes, but also presents with cardiac toxicity. In this post-hoc exploratory analysis of the OptiTrain trial, the effects of exercise on cardiotoxicity were monitored by assessing fitness and biomarkers over the intervention and into survivorship. Methods; Women starting chemotherapy were randomized to 16-weeks of resistance and high-intensity interval training (RT-HIIT), moderate-intensity aerobic and high-intensity interval training (AT–HIIT), or usual care (UC). Outcome measures included plasma troponin-T (cTnT), Nt-pro-BNP and peak oxygen uptake (VO2peak), assessed at baseline, post-intervention, and at 1- and 2-years. Results For this per-protocol analysis, 88 women met criteria for inclusion. Plasma cTnT increased in all groups post-intervention. At the 1-year follow-up, Nt-pro-BNP was lower in the exercise groups compared to UC. At 2-years there was a drop in VO2peak for patients with high cTnT ...
Medicine & Science in Sports & Exercise, 2020
The FASEB Journal, 2007
11 subjects performed one-legged exercise program for 5 weeks. The subjects exercised one leg for... more 11 subjects performed one-legged exercise program for 5 weeks. The subjects exercised one leg for 45 min with restricted blood flow, followed by exercise with the other leg at the same absolute workload with unrestricted blood flow. Gene expression of MMP-2, -9, -14 and TIMP-1 were measured in m vastus lateralis before and after the training period, as well as 2 h after the first and last exercise bouts. MMP-14 and MMP-2 both increased robustly with training regardless of training condition, but with no increase after a single exercise bout. MMP-9 increased transiently after a single bout of exercise in the restricted leg, but no increase was evident with training. TIMP-1 had a tendency to increase after a single bout of exercise in the restricted leg and increased with training in both conditions, however more in the restricted leg. This support the importance of MMP for the adaptation and remodelling of the skeletal muscle with exercise training. The difference in expression of MMP-9 and TIMP-1 between ...
Adoptive transfer of anti-tumor cytotoxic T cells is a novel form of cancer immunotherapy, and a ... more Adoptive transfer of anti-tumor cytotoxic T cells is a novel form of cancer immunotherapy, and a key challenge is to ensure the survival and function of the transferred T cells. Immune cell survival requires adaptation to different micro-environments, and particularly to the hypoxic milieu of solid tumors. The HIF transcription factors are an essential aspect of this adaptation, and we undertook experiments to define structural determinants of HIF that would potentiate anti-tumor efficacy in cytotoxic T cells. We created retroviral vectors to deliver ectopic expression of HIF-1ɑ and HIF-2ɑ in mouse CD8+ T cells, together or individually, and with or without sensitivity to their oxygen-dependent inhibitors Von Hippel-Lindau (VHL) and Factor Inhibiting HIF (FIH). We found that HIF-2ɑ, but not HIF-1ɑ, drives broad transcriptional changes in CD8+ T cells, resulting in increased cytotoxic differentiation and cytolytic function against tumor targets. We further found that a specific mutat...
Medicine & Science in Sports & Exercise, 2020
PURPOSE:Regular physical exercise provides a significant risk reduction for breast cancer and rec... more PURPOSE:Regular physical exercise provides a significant risk reduction for breast cancer and recent studies suggest beneficial effects also on disease specific recurrence and mortality. However, little is known about how exercise exerts its protective effects. The primary aim of this study to evaluate the effect of voluntary running on tumor progression and metastasis in the PyMT mouse model of breast cancer. METHODS:From 4 weeks of age, female MMTV-PyMT mice on the FVB background were housed with access either to wirelessly recording running wheels or locked control wheels. Tumor growth was monitored continuously, tumor stage and pulmonary metastases were determined histologically at the 12 week endpoint. In a follow up study, pre-trained female FVB mice were injected intravenously with 2*10PyMT derived tumor cells (IC3) and after an additional 10 weeks of voluntary running, pulmonary metastases and immune cell infiltration was quantified (histologically and with flow cytometry). The CD8+ T-cell population was deleted using weekly administration of CD8 specific antibodies. RESULTS: PyMT mice average running distance was 6.4±2.4 km/day. No significant effects of voluntary running on tumorinitiation, volume or stage were found. However, a reduced number of metastases were observed in mice with access to running wheels (Ctrl 22.0±6.8 and Runners 9.1±1.7). Significant reductions in pulmonary metastasis frequency were also found in runners after intravenous injections of tumor cells (Ctrl 5.2±1.1 and Runners 1.9±0.7) and runningmice had a lower number ofmetastases with a high proliferation score. Metastatic lesions from running mice showed higher content of Granzyme B positive cells (Ctrl 1.2±0.5 and Runners 4.9±1.1), indicating an increased infiltration of cytotoxic T-cells. Depletion of CD8+ cells abolished the reduction in metastatic burden found in running mice compared to non-running mice All data is presented as Mean±SEM. CONCLUSIONS: In this highly aggressive, genetic, breast cancer model, an average of 6 km/day of voluntary running showed little effect on tumor formation and growth. However, the findings suggest that physical activity reduced outgrowth of metastatic lesions through an increased infiltration of cytotoxic immune cells.