Citations

Abstract Genome-Wide Association Studies (GWAS) have begun to investigate associations between inherited genetic variations and breast cancer prognosis. Here, we report our findings from a GWAS conducted in 536 patients with early-onset breast cancer aged 40 or less at diagnosis and with a mean follow-up period of 4.1 years (SD = 1.96). Patients were selected from the Prospective Study of Outcomes in Sporadic versus Hereditary breast cancer. A Bonferroni correction for multiple testing determined that a P value of 1.0 × 10−7 was a statistically significant association signal. Following quality control, we identified 487,496 single nucleotide polymorphisms (SNP) for association tests in stage 1. In stage 2, 35 SNPs with the most significant associations were genotyped in 1,516 independent cases from the same early-onset cohort. In stage 2, 11 SNPs remained associated in the same direction (P ≤ 0.05). Fixed effects meta-analysis models identified one SNP associated at close to genome wide level of significance 556 kb upstream of the ARRDC3 locus [HR = 1.61; 95% confidence interval (CI), 1.33–1.96; P = 9.5 × 10−7]. Four further associations at or close to the PBX1, RORα, NTN1, and SYT6 loci also came close to genome-wide significance levels (P = 10−6). In the first ever GWAS for the identification of SNPs associated with prognosis in patients with early-onset breast cancer, we report a SNP upstream of the ARRDC3 locus as potentially associated with prognosis (median follow-up time for genotypes: CC = 4 years, CT = 3 years, and TT = 2.7 years; Wilcoxon rank-sum test CC vs. CT, P = 4 × 10−4 and CT vs. TT, P = 0.76). Four further loci may also be associated with prognosis. Cancer Res; 73(6); 1883–91. ©2012 AACR.

Bone marrow‐derived macrophages (BMMs) treated with granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) or macrophage colony‐stimulating factor (M‐CSF), differentiate into GM‐CSF‐induced mouse bone marrow‐derived macrophages (GM‐BMMs) or M‐CSF‐induced mouse bone marrow‐derived macrophages (M‐BMMs), which have an M1 or M2 profile, respectively. GM‐BMMs produce large amounts of proinflammatory cytokines and mediate resistance to pathogens, whereas M‐BMMs produce antiinflammatory cytokines that contribute to tissue repair and remodeling. M‐BMMs stimulated with lipopolysaccharide (LPS) are in an antiinflammatory state, with an IL‐12lowIL‐10high phenotype. However, the regulation of this process remains unclear. Klf10 belongs to the family of Krüppel‐like transcription factors and was initially described as a TGF‐β inducible early gene 1. IL‐12p40 is upregulated in LPS‐stimulated M‐BMMs from Klf10‐deficient mice, but downregulated during Klf10 overexpression. Klf11, another member of the Krüppel‐like factor family, can also repress the production of IL‐12p40. Furthermore, Klf10 binds to the CACCC element of the IL‐12p40 promoter and inhibits its transcription. We have therefore identified Klf10 as a transcription factor that regulates the expression of IL‐12p40 in M‐BMMs.

AbstractSTAT3 regulates cell growth by up-regulating downstream targets, such as Myc. The frequency of phosphorylated STAT3 (pSTAT3) and Myc expression and their prognostic relevance is unknown within diffuse large B-cell lymphoma (DLBCL) germinal center B-cell (GCB) and non-GCB subtypes. pSTAT3 and Myc were studied by immunohistochemistry (IHC) on tumors from 40 DLBCL patients uniformly treated on a clinical trial of epratuzumab/rituximab-CHOP. A total of 35% of cases were pSTAT3-positive, and pSTAT3 positivity was more frequent in the non-GCB (P = .06) type but did not correlate with event-free survival (EFS). Myc expression was observed in 50% of cases and was more frequent in non-GCB type (P = .07). Myc-positive cases had inferior EFS in all patients, including the GCB and pSTAT3-positive cases, were more likely to express Myc (P = .06). Myc translocations involving the major breakpoint regions were found in 10% (3 of 29) of cases, and all 3 cases were GCB and had an inferior EFS (P = .09). pSTAT3, but not Myc expression, was correlated with elevated pretreatment serum cytokines, such as IL-10 (P = .05), G-CSF (P = .03), and TNF-α (P = .04). pSTAT3 IHC in DLBCL tumors has the potential to identify patients for STAT3 pathway–directed therapy; Myc IHC is a potential marker for inferior EFS in GCB patients.

Abstract Introduction: The mammalian target of rapamycin (mTOR), a downstream kinase of the PI3K/Akt signaling pathway, is a critical regulator of basic cellular functions, especially in tumor progression, and therefore has emerged as a key target for cancer chemotherapy. Rapamycin and its derivatives (e.g. Everolimus, Temsirolimus and Deferolimus) are mTOR inhibitors originally approved as immunosuppressants to prevent renal and cardiac allograft rejection. These mTOR inhibitors have recently been used in a large number of cancer clinical trials, including renal cell carcinoma, breast cancer, endometrial carcinoma, glioblastoma and mantle cell lymphoma. However, severe side effects such as toxicity and immune suppression limit the escalation of mTOR inhibitors' dosage, resulting in a low responsive rate. To maximize the efficacy and minimize side effects of mTOR inhibitors, there is a critical need to identify the genetic biomarkers for response to mTOR inhibitors and the mechanisms by which they manifest their therapeutic benefits in cancer. Purpose: This study aims to identify and validate novel genetic biomarkers that might influence the therapeutic responses of mTOR inhibitors. Methods: We used 273 human lymphoblastoid cell lines (LCLs), for which we have obtained genome-wide SNP data, Affymetric U133 plus 2 mRNA expression array data, and Illumina microRNA array data, to perform cytotoxicity assays with two mTOR inhibitors, Rapamycin and Everolimus. We then performed genome-wide association (GWA) analyses using SNPs, mRNA gene expression and microRNA data with cytotoxicity data (AUC values) collected on both mTOR inhibitors. Integrated analysis was then performed to identify the SNPs, and microRNAs that might be associated with genes, whose expressions in turn were significantly associated with the cytotoxicity. We then selected candidate genes to functionally validate their effects on sensitivity of mTOR inhibitors using an siRNA screening approach with 3 cell lines, a renal carcinoma (Caki2), a glioblastoma (U87) and a fibroblast (IMR90) cell line, followed by MTS cell proliferation assay and colony formation assay. Results: mRNA expression levels of 48 and 55 genes were associated with Rapamycin and Everolimus cytotoxicity (p<10−4) respectively, with 16 common genes between two drugs. One hundred and twenty seven and 100 SNPs were associated with Rapamycin and Everolimus cytotoxicity (p<10−4), respectively. We selected 23 candidate genes based on the GWA analyses (mRNA expression vs. cytotoxicity, SNPs vs. cytotoxicity and integrated analysis), and verified 13 genes that significantly altered cells' sensitivity to either one or both mTOR inhibitors in at least one cell line based on siRNA screening study. In addition, one microRNA, miR10a, was shown to be most significantly associated with cytotoxicity for both Rapamycin and Everolimus (p=3.3×10−4; p =1.4×10−4). Interestingly, this particular microRNA was also found to be strongly associated with gene expression levels of 9 out 23 selected candidate genes with p<10−4. Conclusions: This study identified a series of novel candidate genes and microRNA (miR10a) that might contribute to the response of mTOR inhibitors. These findings will enhance our understanding of the regulation of mTOR pathway and the mechanisms underlying the variation in response to mTOR inhibitors. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr PR-11.