Development of predictive multimarker test for preeclampsia in early and late pregnancy
Date
2023-06-15
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Abstract
Sõltuvalt maailmajaost ja riigis kättesaadavast tervishoiuteenuse kvaliteedist mõjutab preeklampsia (PE) 3–5% kõigist rasedustest. Selle peamisteks kliinilisteks sümptomiteks on raseduse teisel poolel arenev hüpertensioon ja proteinuuria. PE võib põhjustada emal organpuudulikkust ning ka ema ja/või loote surma. PE-d iseloomustav uteroplatsentaarne puudulikkus põhjustab raskendatud ainevahetust ema ja loote vahel, laguproduktide eemaldamist ja üldist regulatiivset pärsingut. Kliiniliselt eristatakse varajast, enne 34. rasedusnädalat avalduvat ja hilist, alates 34. rasedusnädalast avalduvat PE-d. Tänapäeval peetakse oluliseks varajast PE ennustust aspiriinil põhineva profülaktika alustamiseks ning hilisemas raseduse faasis haiguse kinnitamist või välistamist. Varajane ennustus põhineb ema baasnäitajatel (eelnev raseduste arv, vanus, varasem PE, rass), ultraheli uuringul ja vereseerumist määratavatel PlGF või PAPP-A tasemetest. Kombineeritud riskihinnang võimaldab tuvastada 90% varajastest PE juhtudest, hilise PE korral aga ainult 40%. PE riskihinnang sFlt-1/PlGF kaudu on raseduse teisel poolel efektiivne ainult varajase PE korral. Hilise PE korral on see lähenemine spetsiifiline ainult 75% juhtudel.
Käesoleva doktoritöö eesmärk oli luua uuenduslik multimarker-immuunuuring ja kombineerida selle abil saadud mõõtmiste andmestikust PE ennustusmudelid:
1. Töötada välja uudne Luminex® xMAP-il põhinev immuunuuring 6PLEX PE seoseliste seerumi biomarkerite määramiseks: ADAM12, sENG, leptiin, PlGF, sFlt-1 ja PTX3.
2. PE ennustus oli III trimestril kogutud proovidest kõige parem, kui ennustusmudelisse kaasati viis biomarkerit (sFlt-1, PlGF, ADAM12, sENG, leptiin) koos ema lisa-faktoritega. Nimetatud mudeli PE ennustustäpsus oli 96,5%.
3. I trimestri PE ennustusmudeli väljatöötamisel kasutati masinõppel põhinevat algoritmi, mis tagas parima PE ennustusmudelina 88,2% täpsuse (kaasates ADAM12, PTX3, sFlt-1 ja emapoolsed faktorid). Platsenta FLT1 rs4769613 genotüübi kaasamisel paranes I trimestri PE ennustusmudeli täpsus 93,5%-ni.
Doktoritöö tulemusena töötasin välja kõrge tundlikkuse ja spetsiifilisusega innovaatilise multimarker-immuunuuringu 6PLEX, mis võimaldab ema vereproovi alusel kõrge täpsusega hinnata PE tekkeriski raseduse esimesel ja teisel poolel. Sellise uuringu eeliseks on kuluefektiivsus ja aja kokkuhoid, kuna huvipakkuvad biomarkerid määratakse samaaegselt ühest proovimaterjalist.
Preeclampsia (PE) affects in total close 3-5% pregnancies worldwide, depending on the access and level of local healthcare. PE is defined as mother`s new hypertension after 20 weeks of gestation with most common additional symptom being proteinuria. It can also cause maternal organ dysfunction (hepatic, renal, haematological) and in worst case mother`s/baby`s death. One of the core causes of PE is uteroplacental pathology, leaded by inadequate spiral artery modification and poor villous development. PE is divided based on its symptomatic presence to be early- (before 34th gestational week) or late-onset (from 34th gestational week). Modern practice for PE management holds two concepts - early phase prediction for prophylaxis with aspirin or late pregnancy PE rule-out in-case of PE suspicion. First, general today to assess PE prediction based on FMF model (screening on gestational weeks 11-14) and assessing maternal characteristics and measuring preferably UtA-PI, PAPP-A or PlGF reaches to correct prognosis in 90% of cases with early-onset PE but moderate 40% with late-onset. PE rule-out method, in case of developed symptoms, by assessing sFlt-1/PLGF ratio only works effectively for early-onset situation. Use in case of late-onset increases false-positive PE predictions sharply. The aim of this thesis was to explore the dynamics and value of proposed serum biomarkers of PE in predicting the risk for the disease, and to combine informative biomarkers and clinical data to develop novel PE prediction models applicable either in early or late pregnancy: 1. The novel Luminex® based 6PLEX assay detects six PE related biomarkers ADAM12, PlGF, sENG, sFlt1, PTX3 and leptin in multiplex format with high precision and low intra- and interassay variation. 2. 6PLEX assay based multicomponent modelling (sFlt-1, PlGF, ADAM12, sENG, leptin) enables superior PE prediction in III trimester with 96.5% accuracy. 3. Applying the 6PLEX assay on I trimester, specifically between 10-14 gestational weeks, allows 88.2% precise PE prediction without any false negative cases and 80% specificity (ADAM12, PTX3, sFlt-1). Incorporation of fetal FLT1 rs4769613 data even improves the prediction performance to 93.5%, raising the precificity metrics. This thesis presents innovative experimental research developing and applying multiplex immunoassay measurements of PE related biomarkers in either early or the late pregnancy. Modelling the PE prediction using this novel assay enable timely identification of high-risk pregnancies, application of preventive measures and targeted monitoring of patients. Benefit of such approach is cost-effectiveness and time-savings as multiple biomarkers are detected at the same time in single reaction.
Preeclampsia (PE) affects in total close 3-5% pregnancies worldwide, depending on the access and level of local healthcare. PE is defined as mother`s new hypertension after 20 weeks of gestation with most common additional symptom being proteinuria. It can also cause maternal organ dysfunction (hepatic, renal, haematological) and in worst case mother`s/baby`s death. One of the core causes of PE is uteroplacental pathology, leaded by inadequate spiral artery modification and poor villous development. PE is divided based on its symptomatic presence to be early- (before 34th gestational week) or late-onset (from 34th gestational week). Modern practice for PE management holds two concepts - early phase prediction for prophylaxis with aspirin or late pregnancy PE rule-out in-case of PE suspicion. First, general today to assess PE prediction based on FMF model (screening on gestational weeks 11-14) and assessing maternal characteristics and measuring preferably UtA-PI, PAPP-A or PlGF reaches to correct prognosis in 90% of cases with early-onset PE but moderate 40% with late-onset. PE rule-out method, in case of developed symptoms, by assessing sFlt-1/PLGF ratio only works effectively for early-onset situation. Use in case of late-onset increases false-positive PE predictions sharply. The aim of this thesis was to explore the dynamics and value of proposed serum biomarkers of PE in predicting the risk for the disease, and to combine informative biomarkers and clinical data to develop novel PE prediction models applicable either in early or late pregnancy: 1. The novel Luminex® based 6PLEX assay detects six PE related biomarkers ADAM12, PlGF, sENG, sFlt1, PTX3 and leptin in multiplex format with high precision and low intra- and interassay variation. 2. 6PLEX assay based multicomponent modelling (sFlt-1, PlGF, ADAM12, sENG, leptin) enables superior PE prediction in III trimester with 96.5% accuracy. 3. Applying the 6PLEX assay on I trimester, specifically between 10-14 gestational weeks, allows 88.2% precise PE prediction without any false negative cases and 80% specificity (ADAM12, PTX3, sFlt-1). Incorporation of fetal FLT1 rs4769613 data even improves the prediction performance to 93.5%, raising the precificity metrics. This thesis presents innovative experimental research developing and applying multiplex immunoassay measurements of PE related biomarkers in either early or the late pregnancy. Modelling the PE prediction using this novel assay enable timely identification of high-risk pregnancies, application of preventive measures and targeted monitoring of patients. Benefit of such approach is cost-effectiveness and time-savings as multiple biomarkers are detected at the same time in single reaction.
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