Sample Preparation Variables in Platelet Proteomics for Biomarker Research

Zellner, Maria; Oehler, Rudolf

doi:10.1002/9780470940297.ch3

Cited by 5 publications

(8 citation statements)

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“…This might have an important influence on APP biomarker studies as platelets are easily activated during sample collection. On top of that, anticoagulants strongly affect platelet activation and degranulation, as demonstrated previously [28]. Great care is therefore essential to avoid activation during venipuncture, blood collection, and platelet isolation.…”

Section: Reviewmentioning

confidence: 89%

Platelets, a reliable source for peripheral Alzheimer’s disease biomarkers?

Veitinger

Varga

Guterres

et al. 2014

acta neuropathol commun

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Peripheral biomarkers play an indispensable role in quick and reliable diagnoses of any kind of disease. With the population ageing, the number of people suffering from age-related diseases is expected to rise dramatically over the coming decades. In particular, all types of cognitive deficits, such as Alzheimer’s disease, will increase. Alzheimer’s disease is characterised mainly by coexistence of amyloid plaques and neurofibrillary tangles in brain. Reliable identification of such molecular characteristics antemortem, however, is problematic due to restricted availability of appropriate sample material and definitive diagnosis is only possible postmortem. Currently, the best molecular biomarkers available for antemortem diagnosis originate from cerebrospinal fluid. Though, this is not convenient for routine diagnosis because of the required invasive lumbar puncture. As a consequence, there is a growing demand for additional peripheral biomarkers in a more readily accessible sample material. Blood platelets, due to shared biochemical properties with neurons, can constitute an attractive alternative as discussed here. This review summarises potential platelet Alzheimer’s disease biomarkers, their role, implication, and alteration in the disease. For easy comparison of their performance, the Hedge effect size was calculated whenever data were available.

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Section: Reviewmentioning

confidence: 89%

Platelets, a reliable source for peripheral Alzheimer’s disease biomarkers?

Veitinger

Varga

Guterres

et al. 2014

acta neuropathol commun

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“…The latter must for sure be addressed in the short term to guarantee data are reproducible between labs and not affected by artifacts related to sample preparation. In agreement with this, basic recommendations for blood collection and processing are already in place [1,15]. In addition, recent advances in platelet proteomics technology-with the possibility of carrying out more quantitative studies-will have a positive impact in the field.…”

Section: Maldi-tof/tofmentioning

confidence: 83%

“…At the same time, as it happens in any successful translational research study, there must be a very good communication between the clinicians that obtain the samples and the proteomic researchers since sample preparation is a key issue. For example, in systemic venous blood studies, blood should be extracted from the median cubital vein using a 21-gauge needle, and a light and controlled tourniquet, and collected it preferably in citrate tubes [15]. Hemolysis must be avoided.…”

Section: á Garcíamentioning

confidence: 99%

“…prostacyclin) should not be accepted since there might be changes in the platelet proteome due to platelet preparation artifacts and not to the pathology that is under investigation. Being aware of this issue, our group and others have been actively involved in providing basic guidelines for platelet clinical proteomics research [1,15]. Having all the above in mind, in the last 8 years platelet proteomics has been applied to the study of several platelet-related diseases (summarized in Table 1).…”

Section: Maldi-tof/tofmentioning

confidence: 99%

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Platelet clinical proteomics: Facts, challenges, and future perspectives

García

2016

Proteomics Clinical Apps

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In recent years, proteomics has been applied to platelet clinical research. Platelets are small enucleated cells that play a fundamental role in hemostasis. In a pathological context, unwanted platelet activation is related to various diseases, primarily thrombosis, but also cancer metastasis, inflammation, immunity, and neurodegenerative diseases. The absence of a nucleus is one of the reasons why proteomics can be considered an ideal analytical tool for platelet research. Indeed, platelet proteomics has allowed the identification of many novel signaling proteins and receptors, several of which are being pursued as potential therapeutic targets. Encouraged by this success, several research groups have recently initiated clinical proteomics studies covering diseases where platelets are involved in some way, such as coronary artery disease, storage pool diseases, uremia, cystic fibrosis, and Alzheimer disease. The goal was to identify platelet biomarkers and drug targets that can help to improve the treatment/diagnosis of the disease and provide further mechanistic evidences of the role platelets play in the pathology. The present article will comment on the recent progress of clinical proteomics in the context of platelet research, challenges, and perspectives for the future ahead.

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“…A tensão imposta ao vaso sanguíneo durante a coleta de sangue, e também as etapas de centrifugação no processo de obtenção de plaquetas lavadas, podem ainda contribuir para este fenômeno (Zellner and Oehler, 2011;Parguiña et al, 2012). Além disso, a presença de vesículas extracelulares no sobrenadante de plaquetas não ativadas, que podem ser fisiologicamente liberadas (Flaumenhaft et al, 2009;Østergaard et al, 2012;Parguiña et al, 2012), e de algumas proteínas envolvidas na inibição da ativação plaquetária, também compõem o secretoma de plaquetas não ativadas.…”

Section: Análise Da Fração Proteica Do Sobrenadante De Plaquetas Por unclassified

Caracterização proteômica comparativa da agregação plaquetária induzida pela trombina e pela PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca

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Aos pesquisadores, pós-graduandos, estagiários e funcionários do LETA. Plaquetas são fragmentos celulares anucleados, derivados de megacariócitos, que estão envolvidos em diversos processos fisiológicos e patológicos, como coagulação, inflamação, trombose, aterosclerose, e metástase e angiogênese tumorais. Para executar estas funções, plaquetas ativadas secretam uma fração solúvel de moléculas presentes em seus conteúdos granulares, que passam a interagir com outras moléculas e células adjacentes ao local da injúria, e com os próprios receptores plaquetários. No entanto, os mecanismos que regem a secreção em plaquetas ainda são pouco conhecidos. Neste sentido, o objetivo deste estudo foi analisar comparativamente a agregação de plaquetas ativadas por dois diferentes agonistas enzimáticos: a trombina, um dos mais importantes agonistas plaquetários, e a PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca, que assim como a trombina, ativa plaquetas através dos receptores PAR-1 e PAR-4. Neste estudo foram utilizadas abordagens de espectrometria de massas e de bioinformática para caracterizar alterações nos proteomas do sedimento de plaquetas não ativadas e ativadas, e também, para em paralelo analisar as frações proteicas e peptídicas presentes no sobrenadante (secretoma). Nas análises do sedimento de plaquetas ativadas tanto por PA-BJ quanto por trombina, foi verificada a menor abundância das proteínas PBP, PF4, proteína S, fibronectina, fator V e alfa-1 antitripsina, entre outras, e que também foram identificadas no sobrenadante (secretadas), e o aumento de abundância das proteínas ADAM-10, tromboxano A2 sintase, integrina αIIb, miosina-9 e fosforilase b, que estão diretamente envolvidas na ativação/agregação. Por outro lado, verificamos que na secreção plaquetária induzida por trombina ocorreu o aumento de abundância de proteínas envolvidas na regulação da formação do coágulo, como a proteína S, PAI1 e antitrombina III, sugerindo que nos eventos disparados pela trombina, exista uma regulação rigorosa de sua ação no local da injúria vascular. Já na secreção induzida por PA-BJ, verificamos o aumento significativo das proteínas amiloide beta A4 e do fibrinogênio, envolvidas na ativação/agregação plaquetária, além da liberação e ativação de MMP1, indicando que esta metaloproteinase atue sinergicamente com a PA-BJ para a formação e estabilização do agregado plaquetário. Nas análises do secretoma de plaquetas não ativadas, identificamos pela primeira vez, a presença das proteínas catalase, anidrase carbônica, inibidor de elastase leucocitária e a glicoproteína rica em histidina, que estão envolvidas na inibição e regulação da ativação plaquetária. A análise da fração peptídica do sobrenadante plaquetário permitiu avaliar pela primeira vez o degradoma gerado no processo de agregação por PA-BJ e trombina. Ao longo desses dez anos de trabalho e aprendizado no Instituto Butantan eu tenho muito que agradecer! Obrigada a todos!!! "É necessário abrir os olhos e perceber que as coisas boas estão dentro de nós, onde os s...

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Sample Preparation Variables in Platelet Proteomics for Biomarker Research

Cited by 5 publications

References 50 publications

Platelets, a reliable source for peripheral Alzheimer’s disease biomarkers?

Platelets, a reliable source for peripheral Alzheimer’s disease biomarkers?

Platelet clinical proteomics: Facts, challenges, and future perspectives

Caracterização proteômica comparativa da agregação plaquetária induzida pela trombina e pela PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca

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