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Uncovering tumor-stroma inter-relationships using quantitative MALDI-Mass spectrometry imaging

ABSTRACT: To facilitate analysis of protein expression changes in in situ tumors and stroma, we took advantage of a mouse model that permits conditional activation of the Ser-Thr kinase ROCK within mammary tumor cells. In this study, we undertook MALDI-MSI analysis of tissue samples derived from our conditional ROCK mammary tumor model, to quantify in an unbiased manner, the proteomic changes occurring during the progression of mammary cancers in their specific spatial contexts.

INSTRUMENT(S): ultraflex, LTQ

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Breast, Stem Cell

DISEASE(S): Breast Cancer

SUBMITTER: Parul Mittal

LAB HEAD: Manuela Klingler-Hoffmann

PROVIDER: PXD019255 | Pride | 2020-09-07

REPOSITORIES: Pride

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			Action	DRS
	20200116_BC-1C_500ng_FAIMS-60_90min_15cm_DE40.raw	Raw
	20200116_BC-1D_500ng_FAIMS-60_90min_15cm_DE40-2.raw	Raw
	20200116_BC-2A_500ng_FAIMS-60_90min_15cm_DE40-2.raw	Raw
	20200116_BC-3A_500ng_FAIMS-60_90min_15cm_DE40-2.msf	Msf
	20200116_BC-3A_500ng_FAIMS-60_90min_15cm_DE40-2.msfView	Msf

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Project description:Dipyridamole (DPM) is widely used to prevent strokes and vascular thrombosis. Combination therapy of DPM and antimetabolites has shown synergistic anticancer activity. This study investigated the chemopreventive effects of DPM in the mouse mammary tumor virus promoter driven polyoma middle T oncoprotein (MMTV-PyMT) metastatic breast cancer model. We also investigated the effects of DPM on gene and miRNA expression. Chemopreventive activity was assessed by comparing the time to onset of palpable lesions, primary tumor growth kinetics and the number of lung metastases in transgenic mice treated with DPM or vehicle. Gene expression and microRNA (miRNA) expression profiles of mammary tumor tissues were then analyzed using the Affymetrix GeneChipM-BM-. or miRNA 2.0 arrays. Real-time quantitative PCR (qPCR) was used to confirm changes in gene expression. Treatment with DPM beginning at the age of four weeks delayed the onset of palpable lesions, delayed tumor progression and suppressed lung metastasis. Microarray gene expression analysis identified 253 genes differentially expressed between DPM-treated and control mammary tumors. miRNA expression analysis revealed that 53 miRNAs were altered by DPM treatment. The results indicate that DPM has chemoprevention activity against breast cancer tumorigenesis and metastasis in mice. The array analyses provide insights into potential mechanisms of DPMM-bM-^@M-^Ys chemopreventive effects, involving upregulation of several genes and miRNAs known to suppress cancer growth and/or metastasis and downregulation of genes known to promote cancer. Some of these genes have not been previously studied in breast cancer and may serve as novel molecular targets for breast cancer chemoprevention. MMTV-PyMT+ female mice were randomly divided into a DPM treatment group (n=10 mice, total of 100 glands) and a vehicle (control) group (n=7 mice, total of 70 glands). When mice were four weeks old, when hyperplasia lesions begin to develop in the FVB strain (21), DPM (10 mg/kg) or vehicle was injected IP into the peritoneum of each mouse on a 5 days on/2 days off dosing schedule for a total of 11 weeks (week one=injection one), until mice were 15 weeks of age. Each of the 10 mammary tumors per mouse was monitored until the study end point. A total six breast tumor RNA samples were used for microarray analysis, with three form mice treated with DPM and three from mice treated with vehicle.

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Uncovering tumor-stroma inter-relationships using quantitative MALDI-Mass spectrometry imaging

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