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Proteomic profiling of urinary extracellular vesicles differentiates breast cancer patients from healthy women

ABSTRACT: Urinary extracellular vesicles (uEVs) reflect the biological conditions of the producing cells, and can allows us to better understand cancer progression. We have, herein, aimed at quantifying the concentration and at generating the proteomic profile of uEVs in breast cancer (BC) patients as compared to that of healthy controls (CT). The proteomic profiling of the uEVs was facilitated through LC-MS/MS.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Sittiruk Roytrakul

PROVIDER: PXD039122 | JPOST Repository | Sun Dec 31 00:00:00 GMT 2023

REPOSITORIES: jPOST

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			Action	DRS
	uEV_N1_RB2_01_27957.mzXML	Mzxml
	uEV_N1_RB2_01_27958.mzXML	Mzxml
	uEV_N1_RB2_01_27959.mzXML	Mzxml
	uEV_N2_RB3_01_27960.mzXML	Mzxml
	uEV_N2_RB3_01_27961.mzXML	Mzxml

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Project description:The kidney and bladder expresses a variety of powerful defense mechanisms to limit urinary infection including iron scavenging, a process called "nutritional immunity". We previously demonstrated that bladder and kidney epithelia generates the archetype of the urinary iron defense, a lipocalin called NGAL which binds Enterochelin, a bacterial siderophore. However, because urinary bacteria can evade the bacteriostatic effects of NGAL by modifying Enterochelin and by producing additional siderophores, other mechanisms of "nutritional immunity" are anticipated. To study the defense in the kidney, we used a cell type specific method of RNA isolation in mouse (adapted from Gay et al, Oregon). A promiscuous form of phosphoribosyltransferase (UPRT) was cloned into the Rosa26 locus using a floxed-stop design which we activated in a cell specific fashion with Atp6v1b1-Cre. Thiouracil was introduced 12hr and 24hrs after urinary tract infection. Isolation of RNA directly from intercalated revealed synthetic (Npas1-Bmal1-Alas), transport (Hrg1, Flvcr1) and metabolic enzymes (Hmox1) of heme metabolism. Hmox1 was detected in the kidney with a luciferase based reporter (Contag et al, Stanford) after infection. CO production was detected in vivo and in AtpCre-mTmG FACS isolated intercalated cells in vitro using a novel metal-based probe synthesized at Columbia (adapted from Liu et al, China). Urinary bacteria (UPEC) with heme transport mutations were not competitive in the colonization of the kidney but conversely were markedly stimulated by iron, suggesting bacterial-host competition for heme capture and metabolism. In fact, infection upregulated both heme synthetic and metabolic genes, suggesting that CO production is induced by infection. Exposure to CO terminated the growth of UPEC. In sum, we have identified an unusual iron trafficking system in the collecting ducts that mirrors the nutritional requirements of UPEC to achieve pyelonephritis. Many of these components are specific to the intercalated cells, consistent with the notion that these cells defend the urinary tract from infection.

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Proteomic profiling of urinary extracellular vesicles differentiates breast cancer patients from healthy women

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