Search

Supplemantary Video 3, LLC1 cells cocultured with NG2- BMSCs

summarized excel file containing the quantitative raw data and processed values in the manuscript

Supplemantary Video 4, LLC1 cells cocultured with NG2+ Ncadherin KO BMSCs

The bone microenvironment is dynamic and undergoes remodeling in normal and pathologic conditions. Whether such remodeling affects disseminated tumor cells (DTC) and bone metastasis remains poorly understood. Here, we demonstrated that pathologic fractures increase metastatic colonization around the injury. NG2+ cells are a common participant in bone metastasis initiation and bone remodeling in both homeostatic and fractured conditions. NG2+ bone mesenchymal stem/stromal cells (BMSC) often colocalize with DTCs in the perivascular niche. Both DTCs and NG2+ BMSCs are recruited to remodeling sites. Ablation of NG2+ lineage impaired bone remodeling and concurrently diminished metastatic colonization. In cocultures, NG2+ BMSCs, especially when undergoing osteodifferentiation, enhanced cancer cell proliferation and migration. Knockout of N-cadherin in NG2+ cells abolished these effects in vitro and phenocopied NG2+ lineage depletion in vivo. These findings uncover dual roles of NG2+ cells in metastasis and remodeling and indicate that osteodifferentiation of BMSCs promotes metastasis initiation via N-cadherin–mediated cell–cell interaction.Significance:The bone colonization of cancer cells occurs in an environment that undergoes constant remodeling. Our study provides mechanistic insights into how bone homeostasis and pathologic repair lead to the outgrowth of disseminated cancer cells, thereby opening new directions for further etiologic and epidemiologic studies of tumor recurrences.This article is highlighted in the In This Issue feature, p. 247

Supplemantary Video 1, LLC1 cells cocultured with NG2+ BMSCs

Supplementary figures and corresponding figure legends

Immunosuppressive elements within the tumor microenvironment, such as tumor-associated macrophages (TAM), can present a barrier to successful antitumor responses by cytolytic T cells. Here we employed preclinical syngeneic p53 null mouse models of triple-negative breast cancer (TNBC) to develop a treatment regimen that harnessed the immunostimulatory effects of low-dose cyclophosphamide coupled with the pharmacologic inhibition of TAMs using either a small-molecule CSF1R inhibitor or an anti-CSF1R antibody. This therapeutic combination was effective in treating several highly aggressive TNBC murine mammary tumor and lung metastasis models. Single-cell RNA sequencing characterized tumor-infiltrating lymphocytes including Th cells and antigen-presenting B cells that were highly enriched in responders to combination therapy. In one model that exhibited long-term posttreatment tumor regression, high-dimensional imaging techniques identified the close spatial localization of B220+/CD86+-activated B cells and CD4+ T cells in tertiary lymphoid structures that were present up to 6 weeks posttreatment. The transcriptional and metabolic heterogeneity of TAMs was also characterized in two closely related claudin-low/mesenchymal subtype tumor models with differential treatment responses. A murine TAM signature derived from the T12 model was highly conserved in human claudin-low breast cancers, and high expression of the TAM signature correlated with reduced overall survival in patients with breast cancer. This TAM signature may help identify human patients with claudin-low breast cancer that will benefit from the combination of cyclophosphamide and anti-CSF1R therapy. These studies illustrate the complexity of the tumor immune microenvironment and highlight different immune responses that result from rational immunotherapy combinations.Significance:Immunostimulatory chemotherapy combined with pharmacologic inhibition of TAMs results in durable treatment responses elicited by Th cells and B cells in claudin-low TNBC models.

Supplementary Data from Chemotherapy Coupled to Macrophage Inhibition Induces T-cell and B-cell Infiltration and Durable Regression in Triple-Negative Breast Cancer

Immunosuppressive elements within the tumor microenvironment, such as tumor-associated macrophages (TAM), can present a barrier to successful antitumor responses by cytolytic T cells. Here we employed preclinical syngeneic p53 null mouse models of triple-negative breast cancer (TNBC) to develop a treatment regimen that harnessed the immunostimulatory effects of low-dose cyclophosphamide coupled with the pharmacologic inhibition of TAMs using either a small-molecule CSF1R inhibitor or an anti-CSF1R antibody. This therapeutic combination was effective in treating several highly aggressive TNBC murine mammary tumor and lung metastasis models. Single-cell RNA sequencing characterized tumor-infiltrating lymphocytes including Th cells and antigen-presenting B cells that were highly enriched in responders to combination therapy. In one model that exhibited long-term posttreatment tumor regression, high-dimensional imaging techniques identified the close spatial localization of B220+/CD86+-activated B cells and CD4+ T cells in tertiary lymphoid structures that were present up to 6 weeks posttreatment. The transcriptional and metabolic heterogeneity of TAMs was also characterized in two closely related claudin-low/mesenchymal subtype tumor models with differential treatment responses. A murine TAM signature derived from the T12 model was highly conserved in human claudin-low breast cancers, and high expression of the TAM signature correlated with reduced overall survival in patients with breast cancer. This TAM signature may help identify human patients with claudin-low breast cancer that will benefit from the combination of cyclophosphamide and anti-CSF1R therapy. These studies illustrate the complexity of the tumor immune microenvironment and highlight different immune responses that result from rational immunotherapy combinations. Significance: Immunostimulatory chemotherapy combined with pharmacologic inhibition of TAMs results in durable treatment responses elicited by Th cells and B cells in claudin-low TNBC models.

The bone microenvironment is dynamic and undergoes remodeling in normal and pathologic conditions. Whether such remodeling affects disseminated tumor cells (DTC) and bone metastasis remains poorly understood. Here, we demonstrated that pathologic fractures increase metastatic colonization around the injury. NG2+ cells are a common participant in bone metastasis initiation and bone remodeling in both homeostatic and fractured conditions. NG2+ bone mesenchymal stem/stromal cells (BMSC) often colocalize with DTCs in the perivascular niche. Both DTCs and NG2+ BMSCs are recruited to remodeling sites. Ablation of NG2+ lineage impaired bone remodeling and concurrently diminished metastatic colonization. In cocultures, NG2+ BMSCs, especially when undergoing osteodifferentiation, enhanced cancer cell proliferation and migration. Knockout of N-cadherin in NG2+ cells abolished these effects in vitro and phenocopied NG2+ lineage depletion in vivo. These findings uncover dual roles of NG2+ cells in metastasis and remodeling and indicate that osteodifferentiation of BMSCs promotes metastasis initiation via N-cadherin–mediated cell–cell interaction. Significance: The bone colonization of cancer cells occurs in an environment that undergoes constant remodeling. Our study provides mechanistic insights into how bone homeostasis and pathologic repair lead to the outgrowth of disseminated cancer cells, thereby opening new directions for further etiologic and epidemiologic studies of tumor recurrences. This article is highlighted in the In This Issue feature, p. 247

To be worthy of its name, the Security Council will need to engage more deliberately and resolutely on new threats involving cyber and emerging technology, climate and the environment, and global h...

Reseña del libro Crítica y comunicación. Sobre políticas de las imágenes (Valencia: tirant humanidades, 2018) de Víctor Silva Echeto.

Many solid cancers metastasize to the bone and bone marrow (BM). This process may occur even before the diagnosis of primary tumors, as evidenced by the discovery of disseminated tumor cells (DTCs) in patients without occult malignancies. The cellular fates and metastatic progression of DTCs are determined by complicated interactions between cancer cells and BM niches. Not surprisingly, these niches also play important roles in normal biology, including homeostasis and turnover of skeletal and hematopoiesis systems. In this Review, we summarize recent findings on functions of BM niches in bone metastasis (BoMet), particularly during the early stage of colonization. In light of the rich knowledge of hematopoiesis and osteogenesis, we highlight how DTCs may progress into overt BoMet by taking advantage of niche cells and their activities in tissue turnover, especially those related to immunomodulation and bone repair.

Immunosuppressive elements within the tumor microenvironment such as Tumor Associated Macrophages (TAMs) can present a barrier to successful anti-tumor responses by cytolytic T cells. We employed preclinical syngeneic p53 null mouse models of triple negative breast cancer (TNBC) to develop a treatment regimen that harnessed the immunostimulatory effects of low-dose cyclophosphamide coupled with the pharmacologic inhibition of TAMs using either a small molecule CSF1R inhibitor or an anti-CSF1R antibody. This therapeutic combination was used to successfully treat several highly aggressive TNBC murine mammary tumors and lung metastasis. Using this regimen and single cell RNA sequencing we characterized tumor infiltrating lymphocytes (TILs) including helper T cells and antigen-presenting B cells that were highly enriched in good responders to combination therapy. Using high dimensional imaging techniques, we identified the close spatial localization of B220+ CD86+ activated B cells and CD4+ T cells in tertiary lymphoid structures that were present up to 6 weeks post-treatment in one model that also exhibited long-term tumor regression post-treatment. We also characterized the transcriptional and metabolic heterogeneity of TAMS in these two closely related claudin-low/mesenchymal subtype tumor models with differential treatment responses. A murine TAM signature derived from the T12 model is highly expressed and conserved in human claudin-low breast cancers, and high expression of the T12 signature correlated with reduced overall survival. This T12 tumor TAM signature may help identify human claudin-low breast cancer patients that will benefit from the combination of cyclophosphamide and anti-CSF1R therapy. These studies illustrate the complexity of the tumor immune microenvironment and highlight different immune responses that result from rationale combinations of immunotherapy.SignificanceA treatment regimen that harnessed the immunostimulatory effects of cyclophosphamide coupled with the inhibition of CSF1R was used to successfully treat several highly aggressive claudin-low TNBC murine mammary tumors and lung metastasis.

Polyclonal metastases frequently arise from clusters of circulating tumor cells (CTCs). CTC clusters metastasize better than single CTCs, but the underlying molecular mechanisms are poorly understood. Here, we show that polyclonal metastatic seeds exhibit higher resistance to natural killer (NK) cell killing. Using breast cancer models, we observed higher proportions of polyclonal lung metastasis in immunocompetent mice compared with mice lacking NK cells. Depleting NK cells selectively increased monoclonal but not polyclonal metastases, suggesting that CTC clusters are less sensitive to NK-mediated suppression. Transcriptional analyses revealed that clusters have elevated expression of cell–cell adhesion and epithelial genes, which is associated with decreased expression of NK cell activating ligands. Furthermore, perturbing tumor cell epithelial status altered NK ligand expression and sensitivity to NK-mediated killing. Collectively, our findings show that NK cells can determine the fate of CTCs of different epithelial and mesenchymal states, and impact metastatic clonal evolution by favoring polyclonal seeding. Zhang and colleagues report that multicellular clusters of circulating tumor cells are more resistant to killing by natural killer (NK) cells through altered NK ligand expression, resulting in polyclonal metastases.

One of the hallmarks of life is the widespread use of certain essential ribozymes. The ubiquitous ribonuclease P (RNase P) and eukaryotic RNase MRP are essential complexes where a structured, noncoding RNA acts in catalysis. Recent discoveries have elucidated the three-dimensional structure of the ancestral ribonucleoprotein complex, suggested the possibility of a protein-only composition in organelles, and even noted the absence of RNase P in a non-free-living organism. With respect to these last two findings, import mechanisms for RNases P/MRP into mitochondria have been demonstrated, and RNase P is present in organisms with some of the smallest known genomes. Together, these results have led to an ongoing debate regarding the precise definition of how "essential" these ribozymes truly are.

Computer Tomographic Colonography, combined with computer-aided detection (CAD), is a promising emerging technique for colonic polyp analysis. We present a CAD scheme for polyp flagging based on new texture and geometric features that consider both the information in the candidate polyp location and its immediate surrounding area, testing multiple sizes. The proposed algorithm is tested with ground truth data, including flat and small polyps, with very promising results.

La periodontitis agresiva localizada ocasiona destrucción ósea severa alrededor de dientes específicos, por lo que en un mismo paciente se encuentran defectos óseos en los que se pueden realizar procedimientos quirúrgicos enfocados a la reconstrucción de los tejidos periodontales perdidos. Objetivo: Valorar clínica y radiográficamente los resultados de procedimientos quirúrgicos periodontales reconstructivos utilizando autoinjertos óseos intraorales solos o combinados con otros materiales de reconstrucción. Caso clínico: Un paciente con periodontitis agresiva localizada, fue tratado con 4 diferentes procedimientos quirúrgicos reconstructivos: 1) autoinjerto óseo combinado con material aloplástico, 2) autoinjerto óseo solo, 3) autoinjerto óseo con membrana absorbible, y 4) autoinjerto óseo mezclado con plasma rico en factores de crecimiento. Resultados: Los resultados se evaluaron clínica y radiográficamente entre los 9 y los 12 meses postoperatorios. Clínicamente se detectó una disminución en la profundidad al sondeo de 10 a 4 mm como valores extremos, encontrándose recesión gingival ≤ 3 mm en los sitios tratados. Radiográficamente, en todos los defectos se apreció llenado óseo. Conclusión: Los procedimientos quirúrgicos reconstructivos que utilizan autoinjertos óseos intraorales, ya sea solos o combinados con otros materiales de reconstrucción, son una buena alternativa en el tratamiento de los defectos intraóseos periodontales en periodontitis agresiva, ya que permiten mejoría de los parámetros clínicos y radiográficos.

In this paper we propose a monoscopic 2D to stereo 3D conversion system. Our process of producing a stereo 3D system from 2D images requires to estimate a relative depth map of the objects in the image that comprises the real world 3D geometry of the scene initially captured. Subsequently, we map the estimated depth into two perspective image views, left and right, with an artificially synthesized parallax between them. We present a depth estimation method based on measuring focus cues, which consists of a local spatial frequency measurement using multiresolution wavelet analysis and a Lipschitz regularity estimation of significant edges; resulting in a pixel resolution depth map. Based on this relative depth map, the stereo 3D image is synthesized with a method that uses interpolated image row sections to artificially generate parallax in the left and right perspective views, and thus when viewed with a stereo 3D display system induce a sense of stereopsis to the observer.

Como parte de un estudio que pretende cuantificar la disponibilidad de biomasa en un sector de la zona árida chilena, se desarrollan funciones para estimar la biomasa de seis especies consideradas importantes, que son: Acacia caven, Cassia coquimbensis, Colliguaya odorifera, Cordia decandra, Flourenzia thurifera y Lithraea caustica.

Quillaja is one of the most important species growing in the semiarid region of Chile because of the high content of saponin in its bark. The wood is normally used as charcoal. Biomass equations to estimate the weight of total stem bark, commercial bark, stem wood, branches and total tree were developed. Data were obtained in three areas located in the north, centre and south of the Chilean semiarid region. Thirty trees were completely weighed. Individual tree components and total tree weights were regressed against DBH, basal diameter, total height and number of main branches, and a combination of them, using the allometric equation of the form Y = a X b and a multiple linear model, both weighted and unweighted. The best results were obtained with the second kind of model, giving better estimates for total tree and stem bark weights. The poorest estimates were obtained for commercial bark.

El objetivo de este trabajo es determinar las funciones de biomasa que mejor estimen el peso de los árboles, para lo cual se muestrearon y pesaron completamente 50 árboles de Quillay, seleccionados de tres zonas de crecimiento de la especie. Para registrar el peso de estos árboles muestra, se separaron y pesaron en forma independiente los principales componentes que conforman el árbol, es decir, fuste, corteza, ramas y ramillas. Para determinar las mejores funciones de biomasa, se probaron modelos matemáticos lineales y alométricos, procesándolos mediante el método de regresión paso a paso. Las funciones calculadas fueron analizadas y comparadas usando varios indicadores estadísticos.

Funciones de biomasa fueron calculadas para dos importantes especies que crecen en la zona central de Chile: Boldo (Peumus boldus) y Espino (Acacia caven). Para lograr este objetivo, se empleó una metodología de terreno y de análisis matemático que ha sido probado con éxito en otras especies nativas de la zona. Las mejores funciones obtenidas son del tiempo logarítimicas para el caso del Boldo y lineales para el Espino. Considerando las buenas correlaciones y bajos errores obtenidos, estas funciones pueden utilizarse con seguridad para estimar el peso total y de los principales componentes de los árboles de Boldo y Espino.