A Novel Role of Oncostatin M in Invasive Breast Cancer: Induction of Cathepsin D and Lysosomal Trafficking

Oncostatin M (OSM) is an interleukin-6 (IL-6) family cytokine shown to be important in inflammation, hematopoiesis, development and bone homeostasis. Despite its role as a growth suppressor for many cancers, including breast cancer, OSM is currently being studied for its ability to promote tumor invasion and metastasis. Cathepsin D (CTSD) is a lysosomal protease found to be overexpressed and hypersecreted in breast and other cancers. In this study, we found OSM to induce the expression of CTSD protein in human breast cancer cells via the STAT3 and JNK2 pathways. Next, we investigated mechanisms resulting in the increased secretion of CTSD from tumor cells. Previous reports have shown that acidic extracellular pH and cellular transformation stimulate lysosomal trafficking, increase secretion of lysosomal proteases, and increase invasion. In this study, we observed that OSM induced a change in cellular morphology and that CTSD-containing lysosomes traffic to the newly formed cellular protrusions. The trafficking and secretion of CTSD was dependent on Na+/H+ exchanger (NHE) activity and OSM activation of the PI3K and p38 MAPK pathways. OSM induced the secretion of physiologically active CTSD which correlated with lysosomal location. Knockdown of CTSD also prevented an increase in invasive potential, even in the presence of OSM. Together, these results suggest that the expression of CTSD and location of CTSD-containing lysosomes are important aspects of the increase in invasive potential of tumor cells induced by OSM. This study provides further evidence that OSM may be an important therapeutic target for in the early stages of breast cancer metastasis

Improving nursing care in a children’s hospital in rural India

Background: Nursing care quality in developing countries is an ongoing challenge leading to poor patient outcomes. The objective of this study is to evaluate changes in nursing performance providing routine cares following a training program in children’s hospital in Mota Fofalia, Gujarat, India. Methods: The main outcome measure was the proportion of newborns with vital signs and weights obtained by nursing staff before and after a training program. The training program consisted of an in-service reinforced by hands-on management of patient care for 2 weeks. Following the training, the nurses were observed for 2 months. Results: Observation of 138 newborn encounters demonstrated a 29.7% improvement in vital sign monitoring and 88.4% in weight monitoring from the 0% baseline. Conclusion: We observed a moderate improvement in measuring vital signs and a substantial improvement in measuring weights in newborns with the training intervention. For further improvement, continued training, and follow-up is indicated

Antenatal Steroids and Cord Blood T-cell Glucocorticoid Receptor DNA Methylation and Exon 1 Splicing

Antenatal administration of glucocorticoids such as betamethasone (BMZ) during the late preterm period improves neonatal respiratory outcomes. However, glucocorticoids may elicit programming effects on immune function and gene regulation. Here, we test the hypothesis that exposure to antenatal BMZ alters cord blood immune cell composition in association with altered DNA methylation and alternatively expressed Exon 1 transcripts of the glucocorticoid receptor (GR) gene in cord blood CD4+ T-cells. Cord blood was collected from 51 subjects in the Antenatal Late Preterm Steroids Trial: 27 BMZ, 24 placebo. Proportions of leukocytes were compared between BMZ and placebo. In CD4+ T-cells, methylation at CpG sites in the GR promoter regions and expression of GR mRNA exon 1 variants were compared between BMZ and placebo. BMZ was associated with an increase in granulocytes (51.6% vs. 44.7% p = 0.03) and a decrease in lymphocytes (36.8% vs. 43.0% p = 0.04) as a percent of the leukocyte population vs. placebo. Neither GR methylation nor exon 1 transcript levels differed between groups. BMZ is associated with altered cord blood leukocyte proportions, although no associated alterations in GR methylation were observed

Additional file 2: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Figure S1. qPCR standard curve derived from spiking cancer cells into mouse blood. MDA-MB-231 cells were spiked into mouse blood, and DNA was extracted and subjected to qPCR analysis. Specific cell numbers were correlated to CT values and were used to construct a standard curve for the CT values extrapolated from experimental mouse blood. (PPTX 186 kb

Additional file 4: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Figure S3. Deterioration of physical condition in MDATO/OSM tumor-bearing mice treated with TET. a MDATO/OSM tumor-bearing mice treated with tetracycline (+TET) lost, on average, 11.4% of their body weight during TET treatment, compared with −TET mice, which gained an average of 5.5% of their body weight over the same period. b Representative image of mice with MDATO/OSM tumors +TET shows prominent spinal column, muscle wasting, and lack of visible adipose tissue. c Gross morphology of normal (left) and abnormal kidneys (right). Normal kidneys have a distinct border between the medulla and the cortex, with the cortex shown in a darker pink/red color and the medulla shown in a lighter pink color. This indicates that normal blood perfusion was taking place. Abnormal kidneys were either both pale and hypoperfused (middle) or damaged (right), with no clear distinction between the cortex and the medulla. d One hundred percent of mice in the +TET group have abnormal kidney morphologies, whereas only 25% of the mice in the −TET group have abnormal kidneys. **p < 0.01 by Fisher’s exact test. e Sera from mice with abnormal kidneys have a statistically significant higher level of OSM than sera from mice with normal kidneys. Data are expressed as mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 by two-tailed Student’s t test. (ZIP 135 kb

Additional file 3: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Figure S2. Representative OSM staining intensity in IHC. 0 = no staining (image not shown), 1 = light staining, 2 = moderate staining, 3 = heavy staining. (PPTX 27 kb

Additional file 8: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Figure S7. Test of cell line-specific variance in colony-forming assay between 4T1.2-shLacZ and 4T1.2-shOSM2 cell lines. Approximately 10 and 50 cells of 4T1.2-shLacZ or 4T1.2-shOSM2 cells were seeded onto tissue culture plates and were allowed to incubate until colony formation. No significant differences between the cells were detected with ~ 10 cells seeded; however, there was a small but significant increase in the number of colonies with 4T1.2-shOSM2 cells at 50 cells seeded. Data are expressed as mean ± SEM. *p < 0.05 by one-way ANOVA with Bonferroni’s multiple comparisons test. (PPTX 68 kb

Additional file 1: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Table S1. Primer and probe sequences used for qPCR assay for the detection of CTC in blood obtained from tumor-bearing mouse. Table S2. Table format data for Fig. 1B show total number of patients and cores for each stage of breast tissue assessed. Table S3. Mean expression levels are statistically significantly different among cancerous and normal tissues for both stroma and blood vessel endothelium (p<.001). Table S4. Margin status, Her2/neu status and estrogen receptor (ER) status, were revealed by repeated measures analysis. Mean expression levels are statistically significantly different among cancerous, normal, and metastatic tissues for margin status, and ER status. For Her2 status, significant differences were found among cancerous, normal and between 0 and 1 staining intensity for metastatic tissues (p<.001). (DOCX 343 kb

Additional file 5: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Figure S4. OSM is highly expressed in orthotopic 4T1.2 primary mammary tumors in female BALB/c mice. Histology using H&E confirmed the presence of a large primary mammary tumor (T) 32 days after 4T1.2 mouse mammary tumor cell injection into the fourth mammary fat pad of female BALB/c mice. High OSM expression is seen in the tumor, as is background expression in the normal breast connective tissue (CT). OSM expression is shown to be highest in the invasive edge of the tumor (T) closest to the normal breast connective tissue (CT). Control slides with no primary OSM antibody show low background staining. (PPTX 315 kb

Additional file 6: of OSM potentiates preintravasation events, increases CTC counts, and promotes breast cancer metastasis to the lung

Figure S5. qPCR analysis of lung metastases after intracardiac injections. 4T1.2-shLacZ cells and 4T1.2-shOSM2 cells were introduced via intracardiac injection, and qPCR analysis of the lung metastases indicated that the difference between the groups was not significant by two-tailed Student’s t test. (ZIP 60 kb