Blood plasma derived extracellular vesicles (BEVs): particle purification liquid chromatography (PPLC) and proteomic analysis reveals BEVs as a potential minimally invasive tool for predicting response to breast cancer treatment.
Blood plasma derived extracellular vesicles (BEVs): particle purification liquid chromatography (PPLC) and proteomic analysis reveals BEVs as a potential minimally invasive tool for predicting response to breast cancer treatment.
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DOI：
10.1007/s10549-022-06733-x
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发表时间：
2022-11
影响因子：
3.8
通讯作者：
Okeoma, Chioma M.
中科院分区：
文献类型：
--
作者：
Alvarez, Folnetti A.;Kaddour, Hussein;Lyu, Yuan;Preece, Christina;Cohen, Jules;Baer, Lea;Stopeck, Alison T.;Thompson, Patricia;Okeoma, Chioma M.
含有蛋白质的循环血浆源性细胞外囊泡（BEVs）有望作为预测癌症治疗反应的微创生物标志物。本研究的主要目的是建立颗粒纯化液相色谱（PPLC） BEV分离方法的效率和实用性，并评估BEV在预测乳腺癌（BC）患者对新辅助化疗（NAC）的反应中的作用。采用PPLC分离分离bev和非ev污染物，并对17例计划接受NAC的BC患者的bev进行表征。使用LC-MS/MS，我们比较了来自NAC后达到病理完全缓解（pCR）的患者（n=7）（反应者[R]）和未达到pCR的患者（n=10）（非反应者[NR]）的pplc分离bev的蛋白质组。用分离的bev处理管腔MCF7和基底细胞MDA-MB-231 BC细胞，并通过MTT法评估其代谢活性。在接受NAC之前，NR的BEV浓度升高，ζ电位（ζ-电位）为负。在NR的bev中富集了8种蛋白，其中GP1BA （CD42b）、PECAM-1 （CD31）、CAPN1、HSPB1 （HSP27）和ANXA5通过western blot验证。MTT分析显示，来自R和NR患者的bev增加了MCF7和MDA-MB-231 BC细胞的代谢活性，且在接受NR bev治疗的MCF7中，其幅度最高。基于pplc的电动汽车分离为纯电动汽车提供了一种不含大多数污染物的分析前分离过程。我们的研究结果表明，pplc分离的bev和5种相关蛋白可能被建立为化学耐药的预测因子，从而用于鉴定NR以使其免受NAC的毒性作用。
Circulating blood plasma derived extracellular vesicles (BEVs) containing proteins hold promise for their use as minimally invasive biomarkers for predicting response to cancer therapy. The main goal of this study was to establish the efficiency and utility of the particle purification liquid chromatography (PPLC) BEV isolation method and evaluate the role of BEVs in predicting breast cancer (BC) patient response to neoadjuvant chemotherapy (NAC). PPLC isolation was used to separate BEVs from non-EV contaminants and characterize BEVs from 17 BC patients scheduled to receive NAC. Using LC-MS/MS, we compared the proteome of PPLC-isolated BEVs from patients (n=7) that achieved a pathological complete response (pCR) after NAC (responders [R]) to patients (n=10) who did not achieve pCR (non-responders [NR]). Luminal MCF7 and basaloid MDA-MB-231 BC cells were treated with isolated BEVs and evaluated for metabolic activity by MTT assay. NR had elevated BEV concentrations and negative zeta potential (ζ-potential) prior to receipt of NAC. Eight proteins were enriched in BEVs of NR. GP1BA (CD42b), PECAM-1 (CD31), CAPN1, HSPB1 (HSP27), and ANXA5 were validated using western blot. MTT assay revealed BEVs from R and NR patients increased metabolic activity of MCF7 and MDA-MB-231 BC cells and the magnitude was highest in MCF7s treated with NR BEVs. PPLC-based EV isolation provides a preanalytical separation process for BEVs devoid of most contaminants. Our findings suggest that PPLC-isolated BEVs and the five associated proteins may be established as predictors of chemoresistance, and thus serve to identify NR to spare them the toxic effects of NAC.
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影响因子：
14.9
作者：
Babicki S;Arndt D;Marcu A;Liang Y;Grant JR;Maciejewski A;Wishart DS
通讯作者：
Wishart DS
影响因子：
2.2
作者：
Baek, Rikke;Sondergaard, Evo K. L.;Jorgensen, Malene M.
通讯作者：
Jorgensen, Malene M.
影响因子：
8
作者：
Gibert, B.;Hadchity, E.;Diaz-Latoud, C.
通讯作者：
Diaz-Latoud, C.
影响因子：
7.4
作者：
Ding, Lihua;Liu, Li-E;Qu, Lingbo
通讯作者：
Qu, Lingbo
影响因子：
3.4
作者：
Fu, Q;Garnham, CP;Van Eyk, JE
通讯作者：
Van Eyk, JE