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Proteomics analysis of primary mouse sarcomas post radiation therapy

ABSTRACT: - We revealed that inhibition of glutamine metabolism via genetic deletion or pharmacological inhibition of glutaminase (Gls1) radiosensitizes primary sarcomas in vivo. To delineate the potential mechanism(s) of radiosensitization post radiation therapy (RT), we performed proteomic analysis of Gls1+/+ and Gls1fl/flsarcomas 48 hours post 10 Gy RT. We found that 490 and 217 proteins were differentially expressed in Gls1fl/flsarcomas post 0 Gy and 10 Gy compared to Gls1+/+ sarcomas, respectively. Hallmark pathway analysis revealed that Gls1 deletion with or without RT decreased the expression of protein related to proliferation and translation (E2F targets and G2M checkpoint). Interestingly, Gls1 deletion post-RT increased innate immune response marked by elevated interferon-alpha, interferon-gamma, and natural killer (NK) cell responses. Flow cytometry analysis further validated these findings and showed significantly elevated NK cells, but not dendritic and myeloid cells in primary Gls1-deficient sarcomas compared to Gls1-proficient sarcomas post-RT. Collectively, proteomic and flow cytometry datasets suggested that innate immune response is partly accountable for radiosensitizing Gls1-deficient sarcomas.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Alexander Saltzman

LAB HEAD: Patel, Rutulkumar

PROVIDER: PXD050936 | Pride | 2024-06-03

REPOSITORIES: Pride

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			Action	DRS
	20220316_212505_5905_Spectronaut_031622.sne	Other
	20220616_144943_5905_SN16_library_061622.sne	Other
	20220617_173346_5905_SN16_library_061722_Report.xls	Xls
	5905_acetyl_metox_individual_DIA.kit	Other
	5905_acetyl_metox_individual_DIA.xls	Xls

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Project description:Background: The determination of altered expression of genes in specific tumor types and their effect upon cellular processes may create insight in tumorigenesis and help to design better treatments. The Flatcoated retriever is a dog breed with an exceptionally high incidence of histiocytic sarcomas. The breed develops two distinct entities of histiocytic neoplasia, a soft tissue form and a visceral form. Gene expression studies of these tumors has value for comparable human diseases such as histiocytic/dendritic cell sarcoma for which knowledge is difficult to accrue due to their rare occurrence. In addition, such studies may help in the search for genetic aberrations underlying the genetic predisposition in this dog breed. Methods: Microarray analysis and pathway analyses were performed on fresh-frozen tissues obtained from Flatcoated retrievers with localized, soft tissue histiocytic sarcomas (STHS) and disseminated, visceral histiocytic sarcomas (VHS) and on normal canine spleens from various breeds. Expression differences of nine genes were validated with quantitative real-time PCR (qPCR) analyses. Results: QPCR analyses identified the significantly altered expression of nine genes; PPBP, SpiC, VCAM1, ENPEP, ITGAD (down-regulated), and GTSF1, Col3a1, CD90 and LUM (up-regulated) in the comparison of both the soft tissue and the visceral form with healthy spleen. DAVID pathway analyses revealed 24 pathways that were significantly involved in the development of HS in general, most of which were involved in the DNA repair and replication process. Conclusions: This study identified altered expression of nine genes not yet implicated in histiocytic sarcoma manifestations in the dog nor in comparable human histiocytic/dendritic sarcomas. Extraploration of this downside effect of canine inbreeding strategies for the study of similar sarcomas in humans might also lead to the identification of genes related to these rare malignancies in the human. Microarray expression dataset including STHS: Soft Tissue (localized) Histiocytic Sarcoma; VHS: Visceral (disseminated)Histiocytic Sarcoma; together with normal spleen (NS) samples. Comparisons were analysed in dyeswap on a 2-color platform against a common reference sample, consisting of a multitude of canine organs, including liver, spleen, kidney, lung, hart, intestine and bone.

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Proteomics analysis of primary mouse sarcomas post radiation therapy

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