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Aging reduces the pro-tumorigenic potential of bone marrow cells and influences triple-negative breast cancer progression

ABSTRACT: To examine the effects of recombinant granulin on human mammary stromal fibroblasts, we cultured immortalized GFP+ normal human mammary fibroblasts in the presence of recombinant human granulin (1ug/ml) or PBS every 24h for 6 days. To generate GRN-independent CAFs, we injected immortalized GFP+ human mammary fibroblasts, MCF7Ras human breast carcinoma cells, and 20% Matrigel subcutaneously into nude mice. Tumors were allowed to form for a period of 45 days. GFP+ fibroblasts were isolated from tumors by mincing the tumors, dissociating, and culturing in the presence of 1 ug/ml puromycin for ~3-4 weeks. CAF purity was confirmed by ensuring that 100% of the population was GFP-positive.

ORGANISM(S): Homo sapiens

PROVIDER: GSE75333 | GEO | 2016/07/21

SECONDARY ACCESSION(S): PRJNA304091

REPOSITORIES: GEO

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Aging reduces the pro-tumorigenic potential of bone marrow cells and influences triple-negative breast cancer progression

Project description:To examine the effects of recombinant granulin on human mammary stromal fibroblasts, we cultured immortalized GFP+ normal human mammary fibroblasts in the presence of recombinant human granulin (1ug/ml) or PBS every 24h for 6 days. To generate GRN-independent CAFs, we injected immortalized GFP+ human mammary fibroblasts, MCF7Ras human breast carcinoma cells, and 20% Matrigel subcutaneously into nude mice. Tumors were allowed to form for a period of 45 days. GFP+ fibroblasts were isolated from tumors by mincing the tumors, dissociating, and culturing in the presence of 1 ug/ml puromycin for ~3-4 weeks. CAF purity was confirmed by ensuring that 100% of the population was GFP-positive. We cultured normal human mammary fibroblasts in the presence human granulin (1ug/ml) or PBS control every 24h for 6 days. The analysis showen are for 3 control (control) and 3 granulin-treated (PGRN) fibrobast cultures.

2016-07-21 | E-GEOD-75333 | biostudies-arrayexpress

Gene expression profiles of granulin- and control-treated normal human mammary fibroblasts

2010-11-27 | GSE25619 | GEO

Gene expression profiles of granulin- and control-treated normal human mammary fibroblasts

Project description:To examine the effects of recombinant granulin on human mammary stromal fibroblasts, we cultured normal human mammary fibroblasts in the presence of recombinant human granulin (1ug/ml) or PBS every 24h for 6 days. We cultured normal human mammary fibroblasts in the presence human granulin (1ug/ml) or PBS control every 24h for 6 days. The analysis showen are for 3 control (control) and 3 granulin-treated (PGRN) fibrobast cultures.

2010-11-27 | E-GEOD-25619 | biostudies-arrayexpress

Homo sapiens

Project description:Gene expression profiles of granulin- and control-treated normal human mammary fibroblasts

| PRJNA133965 | ENA

Granulins rescue inflammation, lysosome dysfunction, and neuropathology in a mouse model of progranulin deficiency

Project description:In this study, we investigate whether individual granulin subunits derived from the precursor protein progranulin are beneficial. We test this by asking if delivery of a single granulin to the brains of a preclinical model of FTD-GRN Grn-/-mice is sufficient to ameliorate disease-like phenotypes. We performed intracerebroventricular injections of recombinant adeno-associated virus (rAAV2/1) encoding granulin-2, granulin-4, full-length PGRN or GFP in neonatal Grn+/+ and Grn-/- mice. Following AAV injections, mice were aged for 12 months, tissues collected, and analyzed. Quantitative proteomics of the thalamus identified lysosomal function and neuroinflammation as pathways ameliorated by the expression of granulins. Biochemical markers of lysosomal dysfunction, neuroinflammation, and disease-like pathology were assessed via immunoblot and immunohistochemistry. We found that both granulin-2 and granulin-4 rescue markers of lysosomal dysfunction and neuroinflammation in cortical, hippocampal, and thalamic tissue.

2024-10-15 | PXD041095 | Pride

Spatial-temporal tracking of Brca1-driven breast tumorigenesis from sporadic mutant cells with a genetic mosaic mouse model

Project description:We utilized a genetic tool termed Mosaic Analysis with Double Markers (MADM) to achieve a sporadic loss of heterozygosity of Brca1 & Trp53 in mouse mammary epithelial cells and concomitantly label them with GFP. This MADM-based mouse model initiated cancer with sparse GFP+ mutant cells and developed mammary tumors that resemble human disease at pathological, transcriptomic, and genomic levels. This dataset provides bulk RNA sequencing of twelve mammary tumors from MADM Brca1-Trp53 animals.

2023-10-16 | GSE214433 | GEO

Genome-wide analysis of gene expression in murine breast fibroblasts from normal tissue, hyperplasia, adenoma and carcinoma (MMTV-PyMT model)

Project description:Analysis of differences at gene expression level of hpvE6 immortalized fibroblasts isolated from normal mammary glands and from hyperplasia, adenoma and carcinoma stages using the MMTV-PyMT model (FVB/N background). Analysis demonstrated the activation of specific transcriptional programs in fibroblasts from later stages. Total RNA obtained from isolated hpvE6 immortalized fibroblasts from normal mammary gland (NF) and from different stages of tumour development using the MMTV-PyMT murine breast cancer model. Stages were hyperplasia (HpAF), adenoma (AdAF) and carcinoma (CAF). Fibroblasts were seeded in a deformable Matrigel:collagen I matrix and total RNA isolated 72h later.

2013-03-19 | E-GEOD-45256 | biostudies-arrayexpress

Genome-wide analysis of gene expression in murine breast fibroblasts from normal tissue, hyperplasia, adenoma and carcinoma (MMTV-PyMT model)

2013-03-19 | GSE45256 | GEO

Single-cell RNA-seq of de-differentiated mammary adipocytes during lactation

Project description:Adipose tissue in the mammary gland undergoes dramatic remodeling during reproduction. Adipocytes are replaced by mammary alveolar structures during pregnancy and lactation, then reappear upon weaning. Here, we reveal that adipocytes in the mammary gland de-differentiate into Pdgfrα+ preadipocyte- and fibroblast-like cells during pregnancy, and remain de-differentiated during lactation. Upon weaning, de-differentiated fibroblasts proliferate and re-differentiate into adipocytes. In order to determine the molecular signature of these de-differentiated adipocytes in the mammary gland, we compared these cells with classical adipocytes. Using the AdipoChaser-mT/mG system, we pre-labeled mature adipocytes with GFP expression to characterize the features of these de-differentiated adipocytes (Figure 4A), and then purified CD31-/CD45-/PDGFRα+/Tomato+ and CD31-/CD45-/PDGFRα+/GFP+ cells from the stromal vascular fraction (SVF) of lactating mammary gland at the peak of lactation through FACS. Gene expression analyses showed that the CD31-/CD45-/PDGFRα+/Tomato+ cells were indeed enriched with Tomato expression, while the CD31-/CD45-/PDGFRα+/GFP+ cells were enriched with GFP expression (Figure 4C). We then collected CD31-/CD45-/PDGFRα+/GFP+ cells as single cells for subsequent single cell RNA-sequencing analysis (Figure 4D-G, Supplemental. Figure S1A-G). After the flow sorting and single cell RNA amplification, 26 CD31-/CD45-/PDGFRα+/GFP+ cells passed the quality control, and these cells were used for single-cell RNA-sequencing analysis. Due to technical difficulties in sorting single mature white adipocyte through flow cytometry, adipocytes differentiated from the immortalized murine-derived brown pre-adipocyte cell line were used as mature adipocyte control (Pradhan et al., 2017). Additionally, we also included population RNA-seq experiments, i.e. three mature white adipocyte samples, two GFP+, and six GFP- ones.

2018-06-17 | E-MTAB-6818 | biostudies-arrayexpress

Increased RUNX3 expression mediates tumor-promoting ability of human breast cancer-associated fibroblasts

Project description:Cancer-associated fibroblasts (CAFs) are a major stromal component of human breast cancers and often promote tumor proliferation, progression and malignancy. We previously established an experimental CAF (exp-CAF) cell line equipped with a potent tumor-promoting ability. It was generated through prolonged incubation of immortalized human mammary fibroblasts with human breast cancer cells in a tumor xenograft mouse model. Herein, we found that the exp-CAFs highly express Runt-related transcription factor 3 (RUNX3), while counterpart fibroblasts do not. In breast cancer patients, the proportion of RUNX3-positive stromal fibroblast-like cells tends to be higher in cancerous regions than in non-cancerous regions. These findings suggest an association of RUNX3 with CAF characteristics in human breast cancers. To investigate the functional role of RUNX3 in CAFs, the exp-CAFs with or without shRNA-directed knockdown of RUNX3 were implanted with breast cancer cells subcutaneously in immunodeficient mice. Comparison of the resulting xenograft tumors revealed that tumor growth was significantly attenuated when RUNX3 expression was suppressed in the fibroblasts. Consistently, Ki-67 and CD31 immunohistochemical staining of the tumor sections indicated reduction of cancer cell proliferation and microvessel formation in the tumors formed with the RUNX3-suppressed exp-CAFs. These results suggest that increased RUNX3 expression could contribute to the tumor-promoting ability of CAFs through mediating cancer cell growth and neoangiogenesis in human breast tumors.

2023-08-15 | GSE240453 | GEO

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