Regulation of growth, invasion and metabolism of breast ductal carcinoma through CCL2/CCR2 signaling interactions with MET receptor tyrosine kinases.

With over 60,000 cases diagnosed annually in the US, ductal carcinoma in situ (DCIS) is the most prevalent form of early-stage breast cancer. Because many DCIS cases never progress to invasive ductal carcinomas (IDC), overtreatment remains a significant problem. Up to 20% patients experience disease recurrence, indicating that standard treatments do not effectively treat DCIS for a subset of patients. By understanding the mechanisms of DCIS progression, we can develop new treatment strategies better tailored to patients. The chemokine CCL2 and its receptor CCR2 are known to regulate macrophage recruitment during inflammation and cancer progression. Recent studies indicate that increased CCL2/CCR2 signaling in breast epithelial cells enhance formation of IDC. Here, we characterized the molecular mechanisms important for CCL2/CCR2-mediated DCIS progression. Phospho-protein array profiling revealed that CCL2 stimulated phosphorylation of MET receptor tyrosine kinases in breast cancer cells. Co-immunoprecipitation and proximity ligation assays demonstrated that CCL2-induced MET activity depended on interactions with CCR2 and SRC. Extracellular flux analysis and biochemical assays revealed that CCL2/CCR2 signaling in breast cancer cells enhanced glycolytic enzyme expression and activity. CRISPR knockout and pharmacologic inhibition of MET revealed that CCL2/CCR2-induced breast cancer cell proliferation, survival, migration and glycolysis through MET-dependent mechanisms. In animals, MET inhibitors blocked CCR2-mediated DCIS progression and metabolism. CCR2 and MET were significantly co-expressed in patient DCIS and IDC tissues. In summary, MET receptor activity is an important mechanism for CCL2/CCR2-mediated progression and metabolism of early-stage breast cancer, with important clinical implications.

Expression of CCL2/CCR2 signaling proteins in breast carcinoma cells is associated with invasive progression.

Ductal carcinoma in situ (DCIS) is the most common type of pre-invasive breast cancer diagnosed in women. Because the majority of DCIS cases are unlikely to progress to invasive breast cancer, many women are over-treated for DCIS. By understanding the molecular basis of early stage breast cancer progression, we may identify better prognostic factors and design treatments tailored specifically to the predicted outcome of DCIS. Chemokines are small soluble molecules with complex roles in inflammation and cancer progression. Previously, we demonstrated that CCL2/CCR2 chemokine signaling in breast cancer cell lines regulated growth and invasion through p42/44MAPK and SMAD3 dependent mechanisms. Here, we sought to determine the clinical and functional relevance of CCL2/CCR2 signaling proteins to DCIS progression. Through immunostaining analysis of DCIS and IDC tissues, we show that expression of CCL2, CCR2, phospho-SMAD3 and phospho-p42/44MAPK correlate with IDC. Using PDX models and an immortalized hDCIS.01 breast epithelial cell line, we show that breast epithelial cells with high CCR2 and high CCL2 levels form invasive breast lesions that express phospho-SMAD3 and phospho-p42/44MAPK. These studies demonstrate that increased CCL2/CCR2 signaling in breast tissues is associated with DCIS progression, and could be a signature to predict the likelihood of DCIS progression to IDC.

SPECC1L regulates palate development downstream of IRF6.

SPECC1L mutations have been identified in patients with rare atypical orofacial clefts and with syndromic cleft lip and/or palate (CL/P). These mutations cluster in the second coiled-coil and calponin homology domains of SPECC1L and severely affect the ability of SPECC1L to associate with microtubules. We previously showed that gene-trap knockout of Specc1l in mouse results in early embryonic lethality. We now present a truncation mutant mouse allele, Specc1lΔC510, that results in perinatal lethality. Specc1lΔC510/ΔC510 homozygotes showed abnormal palate rugae but did not show cleft palate. However, when crossed with a gene-trap allele, Specc1lcGT/ΔC510 compound heterozygotes showed a palate elevation delay with incompletely penetrant cleft palate. Specc1lcGT/ΔC510 embryos exhibit transient oral epithelial adhesions at E13.5, which may delay shelf elevation. Consistent with oral adhesions, we show periderm layer abnormalities, including ectopic apical expression of adherens junction markers, similar to Irf6 hypomorphic mutants and Arhgap29 heterozygotes. Indeed, SPECC1L expression is drastically reduced in Irf6 mutant palatal shelves. Finally, we wanted to determine if SPECC1L deficiency also contributed to non-syndromic (ns) CL/P. We sequenced 62 Caucasian, 89 Filipino, 90 Ethiopian, 90 Nigerian and 95 Japanese patients with nsCL/P and identified three rare coding variants (p.Ala86Thr, p.Met91Iso and p.Arg546Gln) in six individuals. These variants reside outside of SPECC1L coiled-coil domains and result in milder functional defects than variants associated with syndromic clefting. Together, our data indicate that palate elevation is sensitive to deficiency of SPECC1L dosage and function and that SPECC1L cytoskeletal protein functions downstream of IRF6 in palatogenesis.

Axillary web syndrome among Chilean women with breast cancer: incidence and possible predisposing factors.

PURPOSE: Breast cancer (BC) is the most common cancer in women worldwide. The main treatment for BC is surgery, which involves an axillary procedure that associates with the development of axillary web syndrome (AWS). The incidence of AWS among Chilean women with BC and its possible predisposing factors are currently unknown. Thus, we aimed to (1) determine the incidence of AWS among Chilean women with BC after surgery and (2) identify possible predisposing factors. METHODS: Within 90 days post-surgery, patients were assessed for AWS, i.e., palpable or visible axillary cords in the axillary region extending down from the mid-axilla to the ipsilateral arm. We then computed the odds ratio with 95% confidence interval (OR [95% CI]) for having AWS considering the following predisposing factors: age, body mass index (BMI), number of lymph nodes removed, axillary procedure, days from surgery to the physical therapy assessment, hospital for the surgery, type of breast surgery, and neoadyuvant chemotherapy. RESULTS: AWS was present in 49 out of 107 patients (45.8%). Younger age and lower BMI appeared as the sole predisposing factors for AWS (age, 0.95 [0.91-0.99]; BMI, normal weight 1.00, overweight 0.35 [0.11-1.12], obesity 0.28 [0.08-0.97]). CONCLUSION: The incidence of AWS among Chilean women with BC was 45.8%. Our study also confirms data from previous reports showing that younger age and low BMI are associated with the development of AWS.

Hemoglobina de reticulocito y su importancia en el diagnóstico temprano de anemia ferropénica / Reticulocyte hemoglobin and its importance in early diagnosis of iron deficiency anemia

Resumen Introducción: El contenido de hemoglobina de reticulocitos (CHr), es un parámetro en la biometría hematológica automatizada que proporciona información sobre el contenido de hierro, por ello se ha utilizado como un marcador de la biodisponibilidad del hierro en la eritropoyesis, permite su detección en una etapa temprana de la anemia ferropénica y otras patologías como inflamación crónica, enfermedad renal crónica; además realizar monitoreo de terapias con eritropoyetina y hierro. Objetivo: Exponer la aplicabilidad de la CHr como un parámetro en el diagnóstico precoz de la anemia por deficiencia de hierro, así como su medición e interpretación. Materiales y métodos: Se realizó la revisión de artículos científicos en inglés y español en las bases de datos PubMed, ScienceDirect, LILACS y Medline, usando descriptores validados en Medical Subject Headings (MeSH), considerando periodo de publicabilidad del 80% inferior a 5 años. Resultados: Se describe la importancia, aplicabilidad, determinación e interpretación de este parámetro como biomarcador específico hemático temprano en el diagnóstico de deficiencia de hierro antes de presentarse cambios morfológicos eritroides. Conclusiones: La CHr es un parámetro de gran utilidad en el diagnóstico temprano de anemia ferropénica y otras patologías como deficiencia funcional de hierro, estados de inflamación crónica y enfermedad renal crónica.

Abstract Introduction: The reticulocyte hemoglobin content (CHr) is a parameter in automated hematological biometrics, which can provide information on the iron content. So it has been used as a marker of the bioavailability of iron in the erythropoiesis, it allows its detection at an early stage of iron deficiency anemia and other pathologies such as chronic inflammation, chronic kidney disease; in addition to monitoring therapy with erythropoietin and iron. Objective: To expose the applicability of CHr as a parameter in the early diagnosis of iron deficiency anemia, as well as its measurement and interpretation. Materials and methods: The review of scientific articles in English and Spanish was carried out in the PubMed, ScienceDirect, LILACS and Medline databases, using descriptors validated in Medical Subject Headings (MeSH), considering the publication period of 80% less than 5 years. Results: The importance, applicability, determination and interpretation of this parameter is described as an early specific biomarker in the blood in the diagnosis of iron deficiency before presenting morphological changes occurring during terminal erythroid differentiation. Conclusions: CHr is a very useful parameter in the early diagnosis of iron deficiency anemia and other pathologies such as functional deficiency, chronic inflammation states and chronic renal disease.

Chemokine Signaling Facilitates Early-Stage Breast Cancer Survival and Invasion through Fibroblast-Dependent Mechanisms.

Ductal carcinoma in situ (DCIS) is the most common form of breast cancer, with 50,000 cases diagnosed every year in the United States. Overtreatment and undertreatment remain significant clinical challenges in patient care. Identifying key mechanisms associated with DCIS progression could uncover new biomarkers to better predict patient prognosis and improve guided treatment. Chemokines are small soluble molecules that regulate cellular homing through molecular gradients. CCL2-mediated recruitment of CCR2+ macrophages are a well-established mechanism for metastatic progression. Although the CCL2/CCR2 pathway is a therapeutic target of interest, little is known about the role of CCR2 expression in breast cancer. Here, using a mammary intraductal injection (MIND) model to mimic DCIS formation, the role of CCR2 was explored in minimally invasive SUM225 and highly invasive DCIS.com breast cancer cells. CCR2 overexpression increased SUM225 breast cancer survival and invasion associated with accumulation of CCL2 expressing fibroblasts. CCR2-deficient DCIS.com breast cancer cells formed fewer invasive lesions with fewer CCL2+ fibroblasts. Cografting CCL2-deficient fibroblasts with DCIS.com breast cancer cells in the subrenal capsule model inhibited tumor invasion and survival associated with decreased expression of aldehyde dehydrogenase (ALDH1), a proinvasive factor, and decreased expression of HTRA2, a proapoptotic serine protease. Through data mining analysis, high expression of CCR2 and ALDH1 and low HTRA2 expression were correlated with poor prognosis of breast cancer patients.Implications: This study demonstrates that CCR2 overexpression in breast cancer drives early-stage breast cancer progression through stromal-dependent expression of CCL2 with important insight into prognosis and treatment of DCIS. Mol Cancer Res; 16(2); 296-308. ©2017 AACR.

Targeted gene silencing of CCL2 inhibits triple negative breast cancer progression by blocking cancer stem cell renewal and M2 macrophage recruitment.

Triple negative breast cancers are an aggressive subtype of breast cancer, characterized by the lack of estrogen receptor, progesterone receptor and Her2 expression. Triple negative breast cancers are non-responsive to conventional anti-hormonal and Her2 targeted therapies, making it necessary to identify new molecular targets for therapy. The chemokine CCL2 is overexpressed in invasive breast cancers, and regulates breast cancer progression through multiple mechanisms. With few approaches to target CCL2 activity, its value as a therapeutic target is unclear. In these studies, we developed a novel gene silencing approach that involves complexing siRNAs to TAT cell penetrating peptides (Ca-TAT) through non-covalent calcium cross-linking. Ca-TAT/siRNA complexes penetrated 3D collagen cultures of breast cancer cells and inhibited CCL2 expression more effectively than conventional antibody neutralization. Ca-TAT/siRNA complexes targeting CCL2 were delivered to mice bearing MDA-MB-231 breast tumor xenografts. In vivo CCL2 gene silencing inhibited primary tumor growth and metastasis, associated with a reduction in cancer stem cell renewal and recruitment of M2 macrophages. These studies are the first to demonstrate that targeting CCL2 expression in vivo may be a viable therapeutic approach to treating triple negative breast cancer.

SPECC1L deficiency results in increased adherens junction stability and reduced cranial neural crest cell delamination.

Cranial neural crest cells (CNCCs) delaminate from embryonic neural folds and migrate to pharyngeal arches, which give rise to most mid-facial structures. CNCC dysfunction plays a prominent role in the etiology of orofacial clefts, a frequent birth malformation. Heterozygous mutations in SPECC1L have been identified in patients with atypical and syndromic clefts. Here, we report that in SPECC1L-knockdown cultured cells, staining of canonical adherens junction (AJ) components, ß-catenin and E-cadherin, was increased, and electron micrographs revealed an apico-basal diffusion of AJs. To understand the role of SPECC1L in craniofacial morphogenesis, we generated a mouse model of Specc1l deficiency. Homozygous mutants were embryonic lethal and showed impaired neural tube closure and CNCC delamination. Staining of AJ proteins was increased in the mutant neural folds. This AJ defect is consistent with impaired CNCC delamination, which requires AJ dissolution. Further, PI3K-AKT signaling was reduced and apoptosis was increased in Specc1l mutants. In vitro, moderate inhibition of PI3K-AKT signaling in wildtype cells was sufficient to cause AJ alterations. Importantly, AJ changes induced by SPECC1L-knockdown were rescued by activating the PI3K-AKT pathway. Together, these data indicate SPECC1L as a novel modulator of PI3K-AKT signaling and AJ biology, required for neural tube closure and CNCC delamination.

Variations in starch physicochemical properties from a generation-means analysis study using amylomaize V and VII parents.

GEMS-0067 (PI 643420) maize line is a homozygous mutant of the recessive amylose-extender (ae) allele and an unknown number of high-amylose modifier (HAM) gene(s). GEMS-0067 produces starch with a approximately 25% higher resistant-starch (RS) content than maize ae single-mutant starches. The objective of this study was to understand how the HAM gene(s) affected the RS content and other properties of ae-background starches. Nine maize samples, including G/G, G/F1, G/H, F1/G, F1/F1, F1/H, H/G, H/F1, and H/H with HAM gene-dosages of 100, 83.3, 66.7, 66.7, 50, 33.3, 33.3, 16.7, and 0%, respectively, were produced from self- and intercrosses of GEMS-0067 (G), H99ae (H), and GEMS-0067xH99ae (F1) in a generation-means analysis (GMA) study. RS contents of examined starches were 35.0, 29.5, 28.1, 32.0, 28.2, 29.4, 12.9, 18.4, and 15.7%, respectively, which were significantly correlated with HAM gene-dosage (r = 0.81, p < 0.01). Amylose content, number of elongated starch granules, and conclusion gelatinization temperature increased with the increase in HAM gene-dosage. X-ray diffraction study showed that the relative crystallinity (%) of starch granules decreased with the increase in HAM gene-dosage. The results suggested that the HAM gene-dosage was responsible for changes in starch molecular structure and organization of starch granules and, in turn, the RS formation in the maize ae mutant starch.

La violencia: lo que nos es mas familiar. Relación entre el poder y el cuerpo en la violencia intrafamiliar / Violence: what is most familiar to us. Relationship between power and the body in domestic violence

El conflicto es un elemento inherente al hombre , se encuentra presente dentro de cualquier espacio en el cual éste se desenvuelve, por lo tanto, el conflicto en si mismo no es un obstáculo para el ser humano que lo inhabilita para su desarrollo, sino una parte constituyente de su complejidad individual y social, potenciadora de cambios.

The conflict is an element inherent to man, it is present within any space in which it develops, therefore, the conflict itself is not an obstacle for the human being that disables it for its development, but a part constituent of its individual and social complexity, promoting change.