Project description:Human colorectal cancer organoids were injected intrasplenically to form liver metastasis in mice were treated with the TROP2-targeting antibody-drug conjugate Sacituzumab Govitecan (SG) or non-targeting control IgG1-SN-38 (ADC). Tumors were harvested at different time points: 0 h, 6 h, 24 h, 48 h, 72 h and 120 h} and subjected to single-cell RNA-seq using sample hashtag multiplexing.
Project description:MicroRNAs are important negative regulators of protein coding gene expression, and have been studied intensively over the last few years. To this purpose, different measurement platforms to determine their RNA abundance levels in biological samples have been developed. In this study, we have systematically compared 12 commercially available microRNA expression platforms by measuring an identical set of 20 standardized positive and negative control samples, including human universal reference RNA, human brain RNA and titrations thereof, human serum samples, and synthetic spikes from homologous microRNA family members. We developed novel quality metrics in order to objectively assess platform performance of very different technologies such as small RNA sequencing, RT-qPCR and (microarray) hybridization. We assessed reproducibility, sensitivity, quantitative performance, and specificity. The results indicate that each method has its strengths and weaknesses, which helps guiding informed selection of a quantitative microRNA gene expression platform in function of particular study goals.
Project description:The severe harm of depression to human life has attracted great attention to neurologists, but its pathogenesis is extremely complicated and has not yet been fully elaborated. Here, we provided a new strategy for revealing the specific pathways of abnormal brain glucose catabolism in depression, which from the supply of energy substrates and the evaluation of mitochondrial structure and function. By using stable isotope-resolved metabolomics technique, we discovered the tricarboxylic acid cycle (TCA cycle) is blocked and the gluconeogenesis is abnormally activated in chronic unpredictable mild stress (CUMS) rats. In addition, our results showed an interesting phenomenon that the brain attempted to activate all possible metabolic enzymes in energy-producing pathways, but CUMS rats still exhibited a low TCA cycle activity due to impaired mitochondria. Depression caused mitochondrial structure and function impaired, and then led to abnormal brain glucose catabolism. The combination of the stable isotope-resolved metabolomics and mitochondrial structure and function analysis can accurately clarify the mechanism of depression. The mitochondrial pyruvate carrier and acetyl-CoA maybe the key targets for depression treatment. The strategy provides a unique insight for exploring the mechanism of depression, the discovery of new targets, and the development of ideal novel antidepressants.
Project description:This experiment was performed to elucidate the effects of CDK6 on CALR-mutant Myeloproliferative Neoplasms. VavCre CALRdel52 knockin mice were crossed with full-body Cdk6-/- mice to obtain Cdk6-/- CALRdel52 mice. Bone marrow from mice of the four genotypes Cdk6+/+ CALR+/+ (wild-type), Cdk6+/+ CALRdel52 (CALR-mutant), Cdk6-/- CALR+/+ (Cdk6 knockout) and Cdk6-/- CALRdel52 (CALR-mutant with Cdk6 knockout) was isolated and stained for flow cytometry. To obtain megakaryocyte progenitors (MkPs) from the bone marrow, Lineage (CD11b, Gr-1, Ter119, CD3, CD19), c-Kit, sca-1, CD150 and CD41 surface markers were used. From the gating strategy: Lineage- c-Kit+ sca-1- CD150+ CD41+, MkPs were sorted into round-bottom collection tubes via fluorescence activated cell sorting (FACS). These cells were then used for RNA-isolation and subsequent RNA-Sequencing. The samples were harvested in two batches (Sample dates: \"February\" and \"April\").