Alberto Herrera

Cancer-associated fibroblast and M2 macrophage markers together predict outcome in colorectal cancer patients.

Tumor epithelial cells within a tumor coexist with a complex microenvironment in which a variety of interactions between its various components determine the behavior of the primary tumors. Cancer‐associated fibroblasts (CAF) and M2 macrophages, characterized by high expression of different markers, including α‐SMA, FSP1 and FAP, or CD163 and DCSIGN, respectively, are involved in the malignancy of different tumors. In the present study, expression of the above markers in CAF and M2 macrophages was analyzed using RT‐PCR and immunohistochemistry in the normal mucosa and tumor tissue from a cohort of 289 colorectal cancer patients. Expression of CAF and M2 markers is associated with the clinical outcome of colorectal cancer patients. Moreover, the combination of CAF and M2 markers identifies three groups of patients with clear differences in the progression of the disease. This combined variable could be a decisive factor in the survival of advanced‐stage patients. Taken together, these analyses demonstrate the prognostic involvement of interrelationships between DCSIGN, CD163, α‐SMA, FSP1 and FAP markers in the survival of colon cancer patients.

Functional heterogeneity of cancer-associated fibroblasts from human colon tumors shows specific prognostic gene expression signature.

Purpose: Cancer-associated fibroblasts (CAF) actively participate in reciprocal communication with tumor cells and with other cell types in the microenvironment, contributing to a tumor-permissive neighborhood and promoting tumor progression. The aim of this study is the characterization of how CAFs from primary human colon tumors promote migration of colon cancer cells. Experimental design: Primary CAF cultures from 15 primary human colon tumors were established. Their enrichment in CAFs was evaluated by the expression of various epithelial and myofibroblast specific markers. Coculture assays of primary CAFs with different colon tumor cells were performed to evaluate promigratory CAF-derived effects on cancer cells. Gene expression profiles were developed to further investigate CAF characteristics. Results: Coculture assays showed significant differences in fibroblast-derived paracrine promigratory effects on cancer cells. Moreover, the association between CAFs promigratory effects on cancer cells and classic fibroblast activation or stemness markers was observed. CAF gene expression profiles were analyzed by microarray to identify deregulated genes in different promigratory CAFs. The gene expression signature, derived from the most protumorogenic CAFs, was identified. Interestingly, this “CAF signature” showed a remarkable prognostic value for the clinical outcome of patients with colon cancer. Moreover, this prognostic value was validated in an independent series of 142 patients with colon cancer, by quantitative real-time PCR (qRT-PCR), with a set of four genes included in the “CAF signature.” Conclusions: In summary, these studies show for the first time the heterogeneity of primary CAFs effect on colon cancer cell migration. A CAF gene expression signature able to classify patients with colon cancer into high- and low-risk groups was identified. Clin Cancer Res; 19(21); 5914–26. ©2013 AACR.

Exosomes enriched in stemness/metastatic-related mRNAS promote oncogenic potential in breast cancer

Cancer cells efficiently transfer exosome contents (essentially mRNAs and microRNAs) to other cell types, modifying immune responses, cell growth, angiogenesis and metastasis. Here we analyzed the exosomes release by breast tumor cells with different capacities of stemness/metastasis based on CXCR4 expression, and evaluated their capacity to generate oncogenic features in recipient cells. Breast cancer cells overexpressing CXCR4 showed an increase in stemness-related markers, and in proliferation, migration and invasion capacities. Furthermore, recipient cells treated with exosomes from CXCR4-cells showed increased in the same abilities. Moreover, inoculation of CXCR4-cell-derived exosomes in immunocompromised mice stimulated primary tumor growth and metastatic potential. Comparison of nucleic acids contained into exosomes isolated from patients revealed a “stemness and metastatic” signature in exosomes of patients with worse prognosis. Finally, our data supported the view that cancer cells with stem-like properties show concomitant metastatic behavior, and their exosomes stimulate tumor progression and metastasis. Exosomes-derived nucleic acids from plasma of breast cancer patients are suitable markers in the prognosis of such patients.

Protumorigenic effects of Snail‐expression fibroblasts on colon cancer cells

Snail1 is a transcriptional factor that plays an important role in epithelial–mesenchymal transition and in the acquisition of invasive properties by epithelial cells. In colon tumors, Snail1 expression in the stroma correlates with lower specific survival of cancer patients. However, the role(s) of Snail1 expression in stroma and its association with patients survival have not been determined. We used human primary carcinoma‐associated fibroblasts (CAFs) or normal fibroblasts (NFs) and fibroblast cell lines to analyze the effects of Snail1 expression on the protumorigenic capabilities in colon cancer cells. Snail1 expression was higher in CAFs than in NFs and, as well as α‐SMA, a classic marker of activated CAFs. Moreover, in tumor samples from 50 colon cancer patients, SNAI1 expression was associated with expression of other CAF markers, such as α‐SMA and fibroblast activation protein. Interestingly, coculture of CAFs with colon cells induced a significant increase in epithelial cell migration and proliferation, which was associated with endogenous SNAI1 expression levels. Ectopic manipulation of Snail1 in fibroblasts demonstrated that Snail1 expression controlled migration as well as proliferation of cocultured colon cancer cells in a paracrine manner. Furthermore, expression of Snail1 in fibroblasts was required for the coadjuvant effect of these cells on colon cancer cell growth and invasion when coxenografted in nude mice. Finally, cytokine profile changes, particularly MCP‐3 expression, in fibroblasts are put forward as mediators of Snail1‐derived effects on colon tumor cell migration. In summary, these studies demonstrate that Snail1 is necessary for the protumorigenic effects of fibroblasts on colon cancer cells.

Thumb metacarpophalangeal joint ligament reconstruction with a triangular tendon graft in posttraumatic chronic instability.

UNLABELLEDnThe purpose of this study was to report the results of static triangular ligament reconstruction, in thumb metacarpophalangeal (MCP) joint chronic posttraumatic laxity using a tendon graft with a proximal apex in ten patients. The mean postoperative follow-up was 40.2 months. The mean postoperative thumb MCP joint stress testing was 43° less than before surgery, and 6.5° less than in the non-injured hand. The mean range of flexion was 10.5° lower in the operated thumb than in the contralateral one, and the mean range of extension was 8° lower. Minimal differences in the values of the Kapandji score, grip and key-pinch strength were found. The preoperative pain became an occasional discomfort after surgery. All patients had a subjective sense of stability until final follow-up. All patients returned to their work or daily activities.nnnLEVEL OF EVIDENCE IVnFor therapeutic studies investigating the results of treatment.

Endothelial cell activation on 3D-matrices derived from PDGF-BB-stimulated fibroblasts is mediated by Snail1

Carcinomas, such as colon cancer, initiate their invasion by rescuing the innate plasticity of both epithelial cells and stromal cells. Although Snail is a transcriptional factor involved in the Epithelial-Mesenchymal Transition, in recent years, many studies have also identified the major role of Snail in the activation of Cancer-Associated Fibroblast (CAF) cells and the remodeling of the extracellular matrix. In CAFs, Platelet-derived growth factor (PDGF) receptor signaling is a major functional determinant. High expression of both SNAI1 and PDGF receptors is associated with poor prognosis in cancer patients, but the mechanism(s) that underlie these connections are not understood. In this study, we demonstrate that PDGF-activated fibroblasts stimulate extracellular matrix (ECM) fiber remodeling and deposition. Furthermore, we describe how SNAI1, through the FAK pathway, is a necessary factor for ECM fiber organization. The parallel-oriented fibers are used by endothelial cells as “tracks”, facilitating their activation and the creation of tubular structures mimicking in vivo capillary formation. Accordingly, Snail1 expression in fibroblasts was required for the co-adjuvant effect of these cells on matrix remodeling and neoangiogenesis when co-xenografted in nude mice. Finally, in tumor samples from colorectal cancer patients a direct association between stromal SNAI1 expression and the endothelial marker CD34 was observed. In summary, our results advance the understanding of PDGF/SNAI1-activated CAFs in matrix remodeling and angiogenesis stimulation.

Differential distribution and enrichment of non-coding RNAs in exosomes from normal and Cancer-associated fibroblasts in colorectal cancer

Exosome production from cancer-associated fibroblasts seems to be an important driver of tumor progression. We report the first in-depth biotype characterization of ncRNAs, analyzed by Next Generation Sequencing and Bioinformatics, expressed in established primary human normal and cancer-associated fibroblasts (CAFs) from cancer and normal mucosa tissues from 9 colorectal cancer patients, and/or packaged in their derived exosomes. Differential representation and enrichment analyses based on these ncRNAs revealed a significant number of differences between the ncRNA content of exosomes and the expression patterns of the normal and cancer-associated fibroblast cells. ncRNA regulatory elements are specifically packaged in CAF-derived exosomes, supporting a specific cross-talk between CAFs and colon cancer cells and/or other stromal cells, mediated by exosomes. These sncRNAs are potential biomarkers present in cancer-associated fibroblast-derived exosomes, which should thereby contribute to developing new non-invasive diagnostic, prognostic and predictive methods for clinical applications in management of cancer patients.

181 Determination of Snail1 Paracrine Functions – Implication in Pro-tumorogenic Abilities on Colorectal Epithelial Cells Lines

180 Molecular Role of EGFR-mediated Docetaxel Resistance in Human Androgen-Independent Prostate Cancer S. Chuang, T. Hour, W. Kang, C. Huang, A.M.E.I. Huang, G. Liu, S. Huang, Y. Pu. Chung Shan Medical University, Immunology, Taichung City, Taiwan, Kaohsiung Medical University, Biochemistry, Kaohsiung City, Taiwan, Kuo General Hospital, Clinical Pathology, Tainan City, Taiwan, National Taiwan University Hospital, Urology, Taipei City, Taiwan, Kaohsiung Medical Hospital, Urology, Kaohsiung City, Taiwan

Comportamiento biomecánico de los metatarsianos durante la marcha en rampa. Distribución de las presiones plantares

The aim of this paper is to study the behaviour of the metatarsals in healthy individuals while walking uphill and downhill through the use of the podometric insole system Pdm 240. We have studied the walking temporal patterns and the pressures registered on the hindfoot and the forefoot, and, within the latter, on each of its metatarsal heads, during the stride and its phases, while the individuals walked on a treadmill at 3.5 Km/h on a 15% gradient. We found that the hindfoot bears more pressure while walking downhill whereas the forefoot bears it walking uphill. If we analyse each metatarsal separately we will see that the highest pressure values are registered when walking uphill. The location of these peaks varies according to the different phases within stance: while walking uphill, they are observed on the midstance phase, and while walking downhill, on that same phase (M1, M4 and M5) and on the forefoot contact phase (M2 and M3). During stance, M1 bears a bigger share of the pressure on a positive gradient, whereas M5 bear it on a negative gradient.