Project description:Breast cancer is a major cause of cancer-associated mortality in females. Although therapeutic strategies targeting subtype-specific receptors have been developed for various breast cancer subtypes, effective targeted therapies for triple-negative breast cancer (TNBC) are lacking. Recently, the role of the tumor microenvironment (TME), particularly cancer-associated fibroblasts (CAFs), in the progression and metastasis of breast cancer, especially TNBC, has attracted increasing attention. However, the role of exosomes secreted by CAFs in TNBC cell proliferation is poorly understood. Thus, in this study, we analyzed exosomes derived from both CAFs and MDA-MB-231 cells and identified exosomal miR-1290 as a key component. When delivered to TNBC cells (BT-549 and Hs578T), exosomal miR-1290 directly suppressed the expression of the putative tumor suppressor TBC1D4, which was accompanied by reduced TSC1 expression and activation of the mTOR pathway. The promotion of TNBC cell growth by miR-1290 through direct targeting of TBC1D4 was validated using 2D culture models, 3D spheroid models, and in vivo xenograft models. Furthermore, we demonstrated that treatment with the mTOR inhibitor rapamycin and the FDA-approved rapamycin analog everolimus effectively suppressed miR-1290-induced TNBC cell proliferation, suggesting a potential therapeutic strategy for TNBC. Moreover, in patient cohorts, elevated levels of circulating miR-1290 significantly distinguished patients with breast cancer from healthy individuals and correlated with poor survival outcomes. Therefore, miR-1290 and its direct target TBC1D4 have potential value in both targeted treatment and diagnosis of TNBC.
Project description:Lipidomic LC/MS of neurons and lipid droplets isolated from mouse. Extracted using MTBE and run on an Agilent 1290 Infinity II LC with MS acquisition on an Agilent QToF 6546.
Project description:Metabolic profiling of serum samples were performed on an Agilent 1290 infinity system (Agilent technologies, Santa-Clara, California, USA) coupled to an AB SCIEX Triple TOF 6600 System (AB SCIEX, Framingham, MA, USA) with an electrospray ion (ESI) source in both positive and negative ion modes. There were 30 differential metabolites under positive ion mode and 23 differential metabolites under negative ion mode, respectively.
Project description:Metastasis accounts for almost 90% of breast cancer-related fatalities, making it frequent malignancy and the main reason of tumor mortality globally among women. A key player in breast cancer is the histone demethylase lysine-specific demethylase 1 (LSD1). We used LSD1 knockdown MCF7 and T47D cell exosomes to treat breast cancer cells for greatly increasing the invasion and migration of breast cancer cells for evaluating the impact of LSD1 on breast cancer invasion and migration. miR-1290 expression was downregulated in LSD1 knockdown MCF7 exosomes. Furthermore, miR-1290 could control NAT1 expression by looking through the database of miR-1290 target genes. These data provide fresh insights into the biology of breast cancer therapy by demonstrating how the epigenetic factor LSD1 stimulates the breast cancer cells’ invasion and migration via controlling exosomal miRNA.
Project description:Metastatic castration-resistant prostate cancer (mCRPC) is a lethal stage of disease for which current biomarkers have modest predictive power. Liquid biopsies have emerged as a minimally invasive approach but may be inaccessible to patients with limited access to clinical facilities. Here we evaluate small-volume dried capillary blood as a biospecimen for liquid biopsies that stabilizes biomarkers for use in decentralized clinical scenarios. We focus on exosomal microRNAs miR-1290 and miR-375 that in venous blood draws predict overall progression and survival in mCRPC. Using a combination of RNA sequencing (RNA-seq) and reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in matched plasma and dried blood samples from 62 mCRPC patients, microRNA recovery was optimized to exceed 50% for abundant microRNAs (miR-16-5p) but was more variable (30–70%) for lower-abundance targets (miR-1290, miR-30a-5p, miR-375). For predictive biomarkers, RT-qPCR consistently measured miR-1290 but was insufficiently sensitive to consistently detect less abundant miR-375 which was selectively depleted from DBS samples relative to plasma. Normalization by abundant, stable miRs was necessary to reduce systematic bias for correlation between plasma and dried blood extracts. Kaplan-Meier analysis showed that high miR-1290/miR-16-5p ratios from dried blood predicted poor survival with a hazard ratio of 3.29 (95% CI = 1.64–6.62, p = 0.0013), performing comparably to PSA. These findings indicate that small-volume dried capillary blood may be a valid biospecimen for decentralized liquid biopsies applying microRNA profiling, and may enhance the flexibility of prognostic testing in mCRPC.
Project description:The extracted MRM peaks from LC-MS/MS experiments were integrated using Agilent MassHunter Quantitative Data Analysis software. Data extraction for GC-MS experiments was performed using Agilent MassHunter Profinder software.