A. Rouzaut | Universidad de Navarra (original) (raw)

Papers by A. Rouzaut

FEBS Journal, 2006

Neurons are highly polarized cells composed of two structurally and functionally distinct parts, ... more Neurons are highly polarized cells composed of two structurally and functionally distinct parts, the axon and the dendrite. The establishment of this asymmetric structure is a tightly regulated process. In fact, alterations in the proteins involved in the configuration of the microtubule lattice are frequent in neuro-oncologic diseases. One of these cytoplasmic mediators is the protein known as collapsin response mediator protein-2, which interacts with and promotes tubulin polymerization. In this study, we investigated collapsin response mediator protein-2 transcriptional regulation during all-trans-retinoic acid-induced differentiation of SH-SY5Y neuroblastoma cells. All-trans-retinoic acid is considered to be a potential preventive and therapeutic agent, and has been extensively used to differentiate neuroblastoma cells in vitro. Therefore, we first demonstrated that collapsin response mediator protein-2 mRNA levels are downregulated during the differentiation process. After completion of deletion construct analysis and mutagenesis and mobility shift assays, we concluded that collapsin response mediator protein-2 basal promoter activity is regulated by the transcription factors AP-2 and Pax-3, whereas E2F, Sp1 and NeuroD1 seem not to participate in its regulation. Furthermore, we finally established that reduced expression of collapsin response mediator protein-2 after all-trans-retinoic acid exposure is associated with impaired Pax-3 and AP-2 binding to their consensus sequences in the collapsin response mediator protein-2 promoter. Decreased attachment of AP-2 is a consequence of its accumulation in the cytoplasm. On the other hand, Pax-3 shows lower binding due to all-trans-retinoic acid-mediated transcriptional repression. Unraveling the molecular mechanisms behind the action of all-trans-retinoic acid on neuroblastoma cells may well offer new perspectives for its clinical application.

IEEE Transactions on Medical Imaging, 2013

We present a fast and robust approach to tracking the evolving shape of whole fluorescent cells i... more We present a fast and robust approach to tracking the evolving shape of whole fluorescent cells in time-lapse series. The proposed tracking scheme involves two steps. First, coherence-enhancing diffusion filtering is applied on each frame to reduce the amount of noise and enhance flow-like structures. Second, the cell boundaries are detected by minimizing the Chan-Vese model in the fast level set-like and graph cut frameworks. To allow simultaneous tracking of multiple cells over time, both frameworks have been integrated with a topological prior exploiting the object indication function. The potential of the proposed tracking scheme and the advantages and disadvantages of both frameworks are demonstrated on 2-D and 3-D time-lapse series of rat adipose-derived mesenchymal stem cells and human lung squamous cell carcinoma cells, respectively.

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1999

Human monocyte subsets, isolated from cultures of mononuclear cells, or freshly obtained from pat... more Human monocyte subsets, isolated from cultures of mononuclear cells, or freshly obtained from patients with multiple sclerosis, Graves`disease or pemphigus vulgaris, differed in phenotype, apoptotic features, mRNA levels of arginase II (A-II) and the inducible form of nitric oxide synthase (iNOS). Liver-type arginase I mRNA was present in all subsets. Apoptosis was followed by the expression of T cell intracellular antigen (TIA) and the simultaneous detection of DNA stainability by propidium iodine and annexin V binding. Apoptosis was practically absent both in activated CD14 CD33 DR CD25 CD69 CD71 a CD16 3 cells, expressing A-II mRNA and having arginase activity, but not iNOS mRNA, and in not fully mature large CD14 CD16 CD23 DR monocytes, expressing simultaneously both mRNAs and having both enzyme activities. However, differentiated small CD14 a CD16 CD69-CD25 a3 CD71 CD23 DR monocytes, expressing high levels of iNOS mRNA, exhibited apoptotic signs. Amounts of NO synthesised by monocytes co-expressing iNOS and arginase changed with the addition of arginine or an iNOS inhibitor; in that case a correlation of NO production and apoptotic features was observed. Data suggest a regulatory role for endogenous NO in apoptosis of stimulated and differentiated monocytes, and also that iNOS and A-II, when simultaneously present, could control the production of NO as a consequence of their competition for arginine.

Los microtubulos desarrollan una funcion importante en procesos celulares tan cruciales como la d... more Los microtubulos desarrollan una funcion importante en procesos celulares tan cruciales como la division celular, la adhesion a sustrato, la migracion o el transporte de proteinas y organulos. Durante la mitosis, los microtubulos constituyen el huso mitotico. Debido a su intervencion en la division celular, los microtubulos son dianas moleculares de primera magnitud en las terapias frente al cancer. En los ultimos anos se ha descubierto que es esencial la colaboracion de las Proteinas Asociadas a Microtubulos (MAPs) para que los microtubulos puedan realizar su funcion correctamente. La familia de las proteinas de respuesta a colapsina o CRMPs constituye un ejemplo de este tipo de moleculas. La proteina mediadora de respuesta a colapsina tipo 2 (CRMP-2) es una proteina adaptadora de tubulina que interacciona con los heterodimeros de tubulina para unirlos a los microtubulos crecientes, de modo dependiente de su estado de fosforilacion. A pesar de su interaccion con tubulina, CRMP-2 ha...

Japanese Journal of Pharmacology

International Immunology, 2010

staining and reduced LSC number/frequency by serial transplantation. Second, through a comparison... more staining and reduced LSC number/frequency by serial transplantation. Second, through a comparison of global gene expression profiles of LSCs and normal HSCs, we identified LSC-specific genes that can potentially be translated into the development of LSC-targeted therapies. Humanized mouse recapitulating human AML and modeling therapy may be useful for translational medicine in the hematologic malignancy.

Cancer research, Oct 22, 2016

Preclinical and clinical evidence indicate that the proimmune effects of radiotherapy can be syne... more Preclinical and clinical evidence indicate that the proimmune effects of radiotherapy can be synergistically augmented with immunostimulatory monoclonal antibodies (mAb) to act both on irradiated tumor lesions and on distant, non-irradiated tumor sites. The combination of radiotherapy with immunostimulatory anti-PD1 and anti-CD137 mAbs was conducive to favorable effects on distant non-irradiated tumor lesions as observed in transplanted MC38 (colorectal cancer), B16OVA (melanoma) and 4T1 (breast cancer) models. The therapeutic activity was crucially performed by CD8 T cells, as found in selective depletion experiments. Moreover, the integrities of BATF-3-dependent dendritic cells specialized in crosspresentation/crosspriming of antigens to CD8+ T cells and of the type I interferon system were absolute requirements for the antitumor effects to occur. The irradiation regimen induced immune infiltrate changes in the irradiated and non-irradiated lesions featured by reductions in the to...

European Journal of Cancer Supplements, 2008

suppression did not decrease the proliferation rate in vitro,rather a slight increase was observe... more suppression did not decrease the proliferation rate in vitro,rather a slight increase was observed. However, tumour growth generated from LDH-A deficient clones was significantly reduced. LDH-B was not increased by shRNA interference for LDH-A in a compensatory mode, while Hif1α expression was increased and PHD2 and CA9 expression were significantly decreased in the LDH-A deficient clones. We show that LDH-A is critical for the growth of colon carcinoma cells in vivo but not in vitro. The LDH-A deficiency seems to induce cellular stress resulting in an increased Hif1α expression and a decreased expression of its regulator, PHD2. A reduced expression of CA9 in those cells may depend on an abrogated lactic acid production. The generation of mouse melanoma (B16F10) and mouse lung carcinoma (Lewis Lung) LDH-A shRNA clones has been successful and the effect in HT-29 cells was reproduced with Lewis lung carcinoma cells but not with B16F10 clones. 85 Poster Analysis of TGFBI overexpression and silencing in the proliferation, migration and chemoresistance of NSCLC cells

Cancer discovery, 2014

Cancer immunotherapy has great promise, but is limited by diverse mechanisms used by tumors to pr... more Cancer immunotherapy has great promise, but is limited by diverse mechanisms used by tumors to prevent sustained antitumor immune responses. Tumors disrupt antigen presentation, T/NK-cell activation, and T/NK-cell homing through soluble and cell-surface mediators, the vasculature, and immunosuppressive cells such as myeloid-derived suppressor cells and regulatory T cells. However, many molecular mechanisms preventing the efficacy of antitumor immunity have been identified and can be disrupted by combination immunotherapy. Here, we examine immunosuppressive mechanisms exploited by tumors and provide insights into the therapies under development to overcome them, focusing on lymphocyte traffic.

European Journal of Cancer Supplements, 2008

Biochemical Journal, 2007

2013 IEEE 10th International Symposium on Biomedical Imaging, 2013

We present a fully automatic approach to quantitatively analyze filopodia-based migration of fluo... more We present a fully automatic approach to quantitatively analyze filopodia-based migration of fluorescent cells in 3D time-lapse series. The proposed method involves three steps. First, each frame of the time-lapse series is preprocessed using a steerable filter and binarized to obtain a coarse segmentation of the cell shape. Second, a sequence of morphological filters is applied on the coarse binary mask to separate the cell body from individual filopodia. Finally, their length is estimated using a geodesic distance transform. The proposed approach is validated on 3D time-lapse series of lung adenocarcinoma cells. We show that the number of filopodia and their average length can be used as a descriptor to discriminate between different phenotypes of migrating cells.

Ejc Supplements, 2008

suppression did not decrease the proliferation rate in vitro,rather a slight increase was observe... more suppression did not decrease the proliferation rate in vitro,rather a slight increase was observed. However, tumour growth generated from LDH-A deficient clones was significantly reduced. LDH-B was not increased by shRNA interference for LDH-A in a compensatory mode, while Hif1α expression was increased and PHD2 and CA9 expression were significantly decreased in the LDH-A deficient clones. We show that LDH-A is critical for the growth of colon carcinoma cells in vivo but not in vitro. The LDH-A deficiency seems to induce cellular stress resulting in an increased Hif1α expression and a decreased expression of its regulator, PHD2. A reduced expression of CA9 in those cells may depend on an abrogated lactic acid production. The generation of mouse melanoma (B16F10) and mouse lung carcinoma (Lewis Lung) LDH-A shRNA clones has been successful and the effect in HT-29 cells was reproduced with Lewis lung carcinoma cells but not with B16F10 clones. 85 Poster Analysis of TGFBI overexpression and silencing in the proliferation, migration and chemoresistance of NSCLC cells

Cancer Discovery, 2012

The tumor microenvironment of transplanted and spontaneous mouse tumors is profoundly deprived of... more The tumor microenvironment of transplanted and spontaneous mouse tumors is profoundly deprived of oxygenation as confi rmed by positron emission tomographic (PET) imaging. CD8 and CD4 tumor-infi ltrating T lymphocytes (TIL) of transplanted colon carcinomas, melanomas, and spontaneous breast adenocarcinomas are CD137 (4-1BB)-positive, as opposed to their counterparts in tumor-draining lymph nodes and spleen. Expression of CD137 on activated T lymphocytes is markedly enhanced by hypoxia and the prolyl-hydroxylase inhibitor dimethyloxalylglycine (DMOG). Importantly, hypoxia does not upregulate CD137 in hypoxia-inducible factor (HIF)-1αknockout T cells, and such HIF-1α-defi cient T cells remain CD137-negative even when becoming TILs, in clear contrast to co-infi ltrating and co-transferred HIF-1α-suffi cient T lymphocytes. The fact that CD137 is selectively expressed on TILs was exploited to confi ne the effects of immunotherapy with agonist anti-CD137 monoclonal antibodies to the tumor tissue. As a result, low-dose intratumoral injections avoid liver infl ammation, achieve antitumor systemic effects, and permit synergistic therapeutic effects with PD-L1/B7-H1 blockade. SIGNIFICANCE: CD137 (4-1BB) is an important molecular target to augment antitumor immunity. Hypoxia in the tumor microenvironment as sensed by the HIF-1α system increases expression of CD137 on tumor-infi ltrating lymphocytes that thereby become selectively responsive to the immunotherapeutic effects of anti-CD137 agonist monoclonal antibodies as those used in ongoing clinical trials. Cancer Discov; 2(7); 608-23. ©2012 AACR.

FEBS Journal, 2006

Neurons are highly polarized cells composed of two structurally and functionally distinct parts, ... more Neurons are highly polarized cells composed of two structurally and functionally distinct parts, the axon and the dendrite. The establishment of this asymmetric structure is a tightly regulated process. In fact, alterations in the proteins involved in the configuration of the microtubule lattice are frequent in neuro-oncologic diseases. One of these cytoplasmic mediators is the protein known as collapsin response mediator protein-2, which interacts with and promotes tubulin polymerization. In this study, we investigated collapsin response mediator protein-2 transcriptional regulation during all-trans-retinoic acid-induced differentiation of SH-SY5Y neuroblastoma cells. All-trans-retinoic acid is considered to be a potential preventive and therapeutic agent, and has been extensively used to differentiate neuroblastoma cells in vitro. Therefore, we first demonstrated that collapsin response mediator protein-2 mRNA levels are downregulated during the differentiation process. After completion of deletion construct analysis and mutagenesis and mobility shift assays, we concluded that collapsin response mediator protein-2 basal promoter activity is regulated by the transcription factors AP-2 and Pax-3, whereas E2F, Sp1 and NeuroD1 seem not to participate in its regulation. Furthermore, we finally established that reduced expression of collapsin response mediator protein-2 after all-trans-retinoic acid exposure is associated with impaired Pax-3 and AP-2 binding to their consensus sequences in the collapsin response mediator protein-2 promoter. Decreased attachment of AP-2 is a consequence of its accumulation in the cytoplasm. On the other hand, Pax-3 shows lower binding due to all-trans-retinoic acid-mediated transcriptional repression. Unraveling the molecular mechanisms behind the action of all-trans-retinoic acid on neuroblastoma cells may well offer new perspectives for its clinical application.

IEEE Transactions on Medical Imaging, 2013

We present a fast and robust approach to tracking the evolving shape of whole fluorescent cells i... more We present a fast and robust approach to tracking the evolving shape of whole fluorescent cells in time-lapse series. The proposed tracking scheme involves two steps. First, coherence-enhancing diffusion filtering is applied on each frame to reduce the amount of noise and enhance flow-like structures. Second, the cell boundaries are detected by minimizing the Chan-Vese model in the fast level set-like and graph cut frameworks. To allow simultaneous tracking of multiple cells over time, both frameworks have been integrated with a topological prior exploiting the object indication function. The potential of the proposed tracking scheme and the advantages and disadvantages of both frameworks are demonstrated on 2-D and 3-D time-lapse series of rat adipose-derived mesenchymal stem cells and human lung squamous cell carcinoma cells, respectively.

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1999

Human monocyte subsets, isolated from cultures of mononuclear cells, or freshly obtained from pat... more Human monocyte subsets, isolated from cultures of mononuclear cells, or freshly obtained from patients with multiple sclerosis, Graves`disease or pemphigus vulgaris, differed in phenotype, apoptotic features, mRNA levels of arginase II (A-II) and the inducible form of nitric oxide synthase (iNOS). Liver-type arginase I mRNA was present in all subsets. Apoptosis was followed by the expression of T cell intracellular antigen (TIA) and the simultaneous detection of DNA stainability by propidium iodine and annexin V binding. Apoptosis was practically absent both in activated CD14 CD33 DR CD25 CD69 CD71 a CD16 3 cells, expressing A-II mRNA and having arginase activity, but not iNOS mRNA, and in not fully mature large CD14 CD16 CD23 DR monocytes, expressing simultaneously both mRNAs and having both enzyme activities. However, differentiated small CD14 a CD16 CD69-CD25 a3 CD71 CD23 DR monocytes, expressing high levels of iNOS mRNA, exhibited apoptotic signs. Amounts of NO synthesised by monocytes co-expressing iNOS and arginase changed with the addition of arginine or an iNOS inhibitor; in that case a correlation of NO production and apoptotic features was observed. Data suggest a regulatory role for endogenous NO in apoptosis of stimulated and differentiated monocytes, and also that iNOS and A-II, when simultaneously present, could control the production of NO as a consequence of their competition for arginine.

Los microtubulos desarrollan una funcion importante en procesos celulares tan cruciales como la d... more Los microtubulos desarrollan una funcion importante en procesos celulares tan cruciales como la division celular, la adhesion a sustrato, la migracion o el transporte de proteinas y organulos. Durante la mitosis, los microtubulos constituyen el huso mitotico. Debido a su intervencion en la division celular, los microtubulos son dianas moleculares de primera magnitud en las terapias frente al cancer. En los ultimos anos se ha descubierto que es esencial la colaboracion de las Proteinas Asociadas a Microtubulos (MAPs) para que los microtubulos puedan realizar su funcion correctamente. La familia de las proteinas de respuesta a colapsina o CRMPs constituye un ejemplo de este tipo de moleculas. La proteina mediadora de respuesta a colapsina tipo 2 (CRMP-2) es una proteina adaptadora de tubulina que interacciona con los heterodimeros de tubulina para unirlos a los microtubulos crecientes, de modo dependiente de su estado de fosforilacion. A pesar de su interaccion con tubulina, CRMP-2 ha...

Japanese Journal of Pharmacology

International Immunology, 2010

staining and reduced LSC number/frequency by serial transplantation. Second, through a comparison... more staining and reduced LSC number/frequency by serial transplantation. Second, through a comparison of global gene expression profiles of LSCs and normal HSCs, we identified LSC-specific genes that can potentially be translated into the development of LSC-targeted therapies. Humanized mouse recapitulating human AML and modeling therapy may be useful for translational medicine in the hematologic malignancy.

Cancer research, Oct 22, 2016

Preclinical and clinical evidence indicate that the proimmune effects of radiotherapy can be syne... more Preclinical and clinical evidence indicate that the proimmune effects of radiotherapy can be synergistically augmented with immunostimulatory monoclonal antibodies (mAb) to act both on irradiated tumor lesions and on distant, non-irradiated tumor sites. The combination of radiotherapy with immunostimulatory anti-PD1 and anti-CD137 mAbs was conducive to favorable effects on distant non-irradiated tumor lesions as observed in transplanted MC38 (colorectal cancer), B16OVA (melanoma) and 4T1 (breast cancer) models. The therapeutic activity was crucially performed by CD8 T cells, as found in selective depletion experiments. Moreover, the integrities of BATF-3-dependent dendritic cells specialized in crosspresentation/crosspriming of antigens to CD8+ T cells and of the type I interferon system were absolute requirements for the antitumor effects to occur. The irradiation regimen induced immune infiltrate changes in the irradiated and non-irradiated lesions featured by reductions in the to...

European Journal of Cancer Supplements, 2008

suppression did not decrease the proliferation rate in vitro,rather a slight increase was observe... more suppression did not decrease the proliferation rate in vitro,rather a slight increase was observed. However, tumour growth generated from LDH-A deficient clones was significantly reduced. LDH-B was not increased by shRNA interference for LDH-A in a compensatory mode, while Hif1α expression was increased and PHD2 and CA9 expression were significantly decreased in the LDH-A deficient clones. We show that LDH-A is critical for the growth of colon carcinoma cells in vivo but not in vitro. The LDH-A deficiency seems to induce cellular stress resulting in an increased Hif1α expression and a decreased expression of its regulator, PHD2. A reduced expression of CA9 in those cells may depend on an abrogated lactic acid production. The generation of mouse melanoma (B16F10) and mouse lung carcinoma (Lewis Lung) LDH-A shRNA clones has been successful and the effect in HT-29 cells was reproduced with Lewis lung carcinoma cells but not with B16F10 clones. 85 Poster Analysis of TGFBI overexpression and silencing in the proliferation, migration and chemoresistance of NSCLC cells

Cancer discovery, 2014

Cancer immunotherapy has great promise, but is limited by diverse mechanisms used by tumors to pr... more Cancer immunotherapy has great promise, but is limited by diverse mechanisms used by tumors to prevent sustained antitumor immune responses. Tumors disrupt antigen presentation, T/NK-cell activation, and T/NK-cell homing through soluble and cell-surface mediators, the vasculature, and immunosuppressive cells such as myeloid-derived suppressor cells and regulatory T cells. However, many molecular mechanisms preventing the efficacy of antitumor immunity have been identified and can be disrupted by combination immunotherapy. Here, we examine immunosuppressive mechanisms exploited by tumors and provide insights into the therapies under development to overcome them, focusing on lymphocyte traffic.

European Journal of Cancer Supplements, 2008

Biochemical Journal, 2007

2013 IEEE 10th International Symposium on Biomedical Imaging, 2013

We present a fully automatic approach to quantitatively analyze filopodia-based migration of fluo... more We present a fully automatic approach to quantitatively analyze filopodia-based migration of fluorescent cells in 3D time-lapse series. The proposed method involves three steps. First, each frame of the time-lapse series is preprocessed using a steerable filter and binarized to obtain a coarse segmentation of the cell shape. Second, a sequence of morphological filters is applied on the coarse binary mask to separate the cell body from individual filopodia. Finally, their length is estimated using a geodesic distance transform. The proposed approach is validated on 3D time-lapse series of lung adenocarcinoma cells. We show that the number of filopodia and their average length can be used as a descriptor to discriminate between different phenotypes of migrating cells.

Ejc Supplements, 2008

suppression did not decrease the proliferation rate in vitro,rather a slight increase was observe... more suppression did not decrease the proliferation rate in vitro,rather a slight increase was observed. However, tumour growth generated from LDH-A deficient clones was significantly reduced. LDH-B was not increased by shRNA interference for LDH-A in a compensatory mode, while Hif1α expression was increased and PHD2 and CA9 expression were significantly decreased in the LDH-A deficient clones. We show that LDH-A is critical for the growth of colon carcinoma cells in vivo but not in vitro. The LDH-A deficiency seems to induce cellular stress resulting in an increased Hif1α expression and a decreased expression of its regulator, PHD2. A reduced expression of CA9 in those cells may depend on an abrogated lactic acid production. The generation of mouse melanoma (B16F10) and mouse lung carcinoma (Lewis Lung) LDH-A shRNA clones has been successful and the effect in HT-29 cells was reproduced with Lewis lung carcinoma cells but not with B16F10 clones. 85 Poster Analysis of TGFBI overexpression and silencing in the proliferation, migration and chemoresistance of NSCLC cells

Cancer Discovery, 2012

The tumor microenvironment of transplanted and spontaneous mouse tumors is profoundly deprived of... more The tumor microenvironment of transplanted and spontaneous mouse tumors is profoundly deprived of oxygenation as confi rmed by positron emission tomographic (PET) imaging. CD8 and CD4 tumor-infi ltrating T lymphocytes (TIL) of transplanted colon carcinomas, melanomas, and spontaneous breast adenocarcinomas are CD137 (4-1BB)-positive, as opposed to their counterparts in tumor-draining lymph nodes and spleen. Expression of CD137 on activated T lymphocytes is markedly enhanced by hypoxia and the prolyl-hydroxylase inhibitor dimethyloxalylglycine (DMOG). Importantly, hypoxia does not upregulate CD137 in hypoxia-inducible factor (HIF)-1αknockout T cells, and such HIF-1α-defi cient T cells remain CD137-negative even when becoming TILs, in clear contrast to co-infi ltrating and co-transferred HIF-1α-suffi cient T lymphocytes. The fact that CD137 is selectively expressed on TILs was exploited to confi ne the effects of immunotherapy with agonist anti-CD137 monoclonal antibodies to the tumor tissue. As a result, low-dose intratumoral injections avoid liver infl ammation, achieve antitumor systemic effects, and permit synergistic therapeutic effects with PD-L1/B7-H1 blockade. SIGNIFICANCE: CD137 (4-1BB) is an important molecular target to augment antitumor immunity. Hypoxia in the tumor microenvironment as sensed by the HIF-1α system increases expression of CD137 on tumor-infi ltrating lymphocytes that thereby become selectively responsive to the immunotherapeutic effects of anti-CD137 agonist monoclonal antibodies as those used in ongoing clinical trials. Cancer Discov; 2(7); 608-23. ©2012 AACR.