IntroductionMicroRNAs (miRNAs) are small non-coding RNA that plays a vital role in cancer progression. Neo-adjuvant chemotherapy (NAC) has become the standard of care for locally advanced breast cancer. The aim of this study was to evaluate miRNA alterations during NAC using multiple samples of tissue and serum to correlate miRNA expression with clinico-pathological features and patient outcomes.MethodsTissue and serum samples were collected from patients with locally advanced breast cancer undergoing NAC at four time points: time of diagnosis, after the first and fourth cycle of doxorubicin/cyclophosphamide treatment, and after the fourth cycle of docetaxel administration. First, we evaluated the miRNA expression profiles in tissue and correlated expression with clinico-pathological features. Then, a panel of four miRNAs (miR-451, miR-3200, miR-21, and miR-205) in serum samples was further validated using quantitative reverse-transcription polymerase chain reaction (RT-qPCR). The alterations in serum levels of miRNA, associations with clinical and pathological responses, correlation with clinico-pathological features, and survival outcomes were studied using Friedman, Mann-Whitney U, and Spearman, Wilcoxon signed-ranks tests. P≤0.05 was considered statistically significant.ResultsWe analyzed 72 tissue samples and 108 serum samples from 9 patients and 27 patients, respectively. MicroRNA expression profiling of tumor versus normal tissue revealed more than 100 differentially expressed miRNAs. Serum miR-451 levels were significantly decreased during treatment, and higher serum levels were associated with improved clinical and pathological responses and disease-free survival. This is one of the early reports on miR-3200 in response to treatment in breast cancer, as serum levels of miR-3200 found to decline during NAC, and higher serum levels were associated with lower residual breast cancer burden and relapse rates at time of diagnosis.ConclusionVariations in serum miRNA levels during NAC treatment may be therapeutically significant for predicting response and survival outcomes.

Acylation stimulating protein (ASP) is a potent lipogenic factor produced from adipocytes. Plasma ASP levels were shown to increase in obesity, diabetes mellitus type II and dyslipidemia, and decrease after weight loss and fasting. Growing evidence suggests that ASP may significantly contribute to subcutaneous fat storage in females. In vitro, ASP stimulated triglyceride synthesis to a larger extent in subcutaneous compared with omental adipocytes. The ASP receptor binding affinity to plasma membranes prepared from adipose tissue showed higher binding affinity to plasma membranes from female adipose tissue compared with male adipose tissue, and was more pronounced to subcutaneous compared with omental plasma membranes. Human studies demonstrated that postprandial triglyceride clearance predicted by ASP levels was more efficient in women than in men. In mice, postprandial triglyceride clearance, with intraperitoneal ASP administration, was faster in females compared with males. The ASP deficient mice were resistant to weight gain and had reduced fat mass that was more pronounced in females. Recent findings in humans and mice point to a significant association between progesterone and ASP variations in females. In this review, we highlight findings, to date, linking ASP to physiological and hormonal alterations that may contribute to subcutaneous fat distribution typical to females.

Objective: The menstrual cycle represents a continuous state of change in terms of female sex steroid environment. Progesterone is linked to increased fat storage while estrogen exerts anti-lipogenic effects. This study investigated variations in the potent lipogenic factor acylation-stimulating protein (ASP), and examined its association with hormonal and lipid profile alterations across the menstrual cycle. Methods and design: Nineteen non-obese women with regular menstrual cycles were investigated in a longitudinal study during the follicular, ovulatory, and mid-luteal phases (ML) of the cycle. Fasting ASP, LH, FSH, progesterone, estradiol, insulin, lipid profile, and apoproteins were evaluated during different phases of the cycle. Results: ASP levels changed significantly throughout the menstrual cycle (K-related Friedman test: PZ0.013). Interestingly, these changes coincide with variations in progesterone levels across the cycle as no significant change in the ASP levels was seen across the follicular phases of the cycle, followed by a significant increase in the ovulatory phase, which continued to elevate toward the ML. The ASP levels correlated positively with the progesterone levels normally elevated in the ML. No significant correlation was seen between ASP and estrogen or any other measured female hormone. Multiple regression analysis including all measured parameters and body mass index showed that progesterone was the only significant predictor of the ASP levels. Conclusion: Our findings suggest that during the menstrual cycle of normal women, the ASP levels coincidentally fluctuate with the progesterone levels, possibly reflecting cooperation between them in fat storage enhancement.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that have unique functions at post-transcriptional level (epigenetics). MiRNAs play a pivotal role in controlling gene expression at various levels including differentiation, cell-cycle regulation, apoptosis and many others in mammals as well as in many organisms. Recently, there has been greater understanding of the contribution of dysregulation of miRNA into disease status in particular carcinogenesis. In this review, we will discuss miRNA discovery, nomenclature, function, contribution of their dysregulation into disease status in particular carcinogenesis and their potential role as biomarkers.

e22099 Background: NAC is used in LABC to downsize the primary tumor and eradicate metastases. This study assessed the Dynamic changes of miRNA and CRP expression during NAC with aim to identify molecular biomarkers that may predict outcome. Methods: Tissue, blood and serum samples were collected at four time points from LABC patients undergoing NAC, including time of diagnosis (base line) after 1st and 4th cycle of Adriamycin/cyclophosphamide, and after 4th cycle of Docetaxel (end point). We used microRNA-microarray for tissue samples and Proximity Ligation Assay (PLA) for serum samples to quantify global miRNA expression and 92 CRPs in LABC patients. The miRNA and CRPS were correlated with clinico-pathological features including stage, grade and clinical response.Statistical analysis was performed using Significant Analysis of Microarray ( SAM), Student's t-test, and Pearson's correlation test. Significant p value was defined as p<0.05. Results: Tissue miRNA expression profiling (n=14) revealed 58 miRNA species that were differentially expressed between NAC endpoint and the baseline (FDR < 15%). Reduced miR-21 expression was associated with disease recurrence. Using PLA (n=33) we identified serumosteoprotegerin, MCP-1, Prolactin, Carbonic Anhydrase IX, CXCL10, CD30L and TGF-alpha levels were significantly associated with overall clinical response (t-test, p<0.05). Moreover, Prolactin and Erythropoietin showed significant associations with the stage (t-test, p<0.05), while MMP3, ErbB4, Midkine, Cathepsin D and Prostasin proteins were associated with the recurrence. Analysis is ongoing to determine the dynamic changes of these biomarkers during the NAC. Conclusions: Our study identified specific miRNAs and CRPs that were differentially expressed during NAC in LABC patients which may have clinical significance.