Project description:The dataset contains data from 12 samples hybridized on Illumina HumanHT-12 array. -patient's primary lymphoma cells obtained from malignant ascites (P2-A1, P2-A2) -patient's non-malignant B-cells (CTRL1, CTRL2) -lymphoma cell line named UPF4D established from the patient's primary lymphoma cells after 15 and 75 days of in vitro cultivation (UPF4D-D15A, UPF4D-D15B, UPF4D-D75) -cells established by xenotransplantation of UPF4D cell line isolated ex vivo from infiltrated murine kidney (UPF4D-K) -cells established by xenotransplantation of UPF4D cell line isolated ex vivo from subcutaneously growing lymphoma (UPF4D-SC) -patient-derived xenograft (PDX) cells named VFN-D2 established by xenotransplantation of primary lymphoma cells into immunodeficient mice isolated ex vivo from the infiltrated murine kidneys (VFN-D2-K1, VFN-D2-K2) -patient-derived xenograft (PDX) cells named VFN-D2 established by xenotransplantation of primary lymphoma cells into immunodeficient mice isolated ex vivo from subcutaneously growing lymphoma (VFN-D2-SC). Samples were sorted using CD19-microbeads (Miltenyi). For xenotransplantation, NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice (referred to as NSG mice) were used.
Project description:Metastasis is the key determinant of poor prognosis for advanced-stage NSCLC patients. Although an important contributor to metastasis is cross-talk between tumor-associated macrophages (TAMs) and tumor cells, its regulation is not fully understood. Expressed primarily in macrophages, scavenger receptor A1 (SR-A1) has been associated with lung tumorigenesis. Here, the mechanistic basis for the involvement of SR-A1 in lung cancer prognosis was investigated using population genetics, transcriptomics, and functional analyses. SR-A1 genetic variants were investigated for possible association with survival of advanced-stage NSCLC patients in the Harvard Lung Cancer Study cohort. Two SNPs (rs17484273, rs1484751) in SR-A1 were significantly associated with poor overall survival of NSCLC patients. Further, data from The Cancer Genome Atlas showed a considerable down-regulation of SR-A1 in lung tumor tissues. The association of SR-A1 with prognosis was validated in animal models in the context of lung cancer metastasis. Macrophages derived from SR-A1 knockout mice accelerated metastasis in a lung cancer mouse model. On the other hand, tumor cell seeding, migration, and invasion as well as macrophage accumulation in lung cancer tissue were enhanced in SR-A1 knockout mice. Furthermore, SR-A1 deficiency promoted up-regulation of serum amyloid A1 (SAA1) in macrophages, which appeared to be mediated by MAPK/Ikappa-B/NF-kappaB signaling. Further, SAA1 exposure promoted tumor cell invasion and macrophage migration in vitro and in vivo, but these effects were blocked by administration of an anti-SAA1 antibody. These findings suggest that SR-A1 may suppress lung cancer metastasis by down-regulating SAA1 production in macrophages.
Project description:Generation of serotonin neurons (SNs) from human pluripotent stem cells (hPSCs) provides a promising platform to explore the mechanisms of serotonin-associated neuropsychiatric disorders. However, neural differentiation always yields heterogeneous cell populations, making it difficult to identify and purify SNs in vitro or track them in vivo following transplantation. Herein, we generated a TPH2-EGFP reporter hPSC line with insertion of EGFP into the endogenous tryptophan hydroxylase 2 (TPH2) locus using CRISPR-Cas9-mediated gene editing technology. This TPH2-reporter, which faithfully indicated TPH2 expression during differentiation, enabled us to obtain purified SNs for subsequent transcriptional analysis and study of pharmacological responses to antidepressants. In addition, the reporter system showed strong EGFP expression to indicate SNs, which enabled us to explore in vitro and ex vivo electrophysiological properties of SNs. In conclusion, this TPH2-EGFP reporter cell line might be of great significance for studies on human SN-related development and differentiation, drug screening, disease modeling, and cell replacement therapies.