Jacob, Jaison, Ngo, Debby, Finkel, Nancy, Pitts, Rebecca, Gleim, Scott, Farrell, Laurie, Keyes, Michelle, Benson, Mark, Morgan, Thomas, Jennings, Lori and Gerszten, Robert (2017) Application of Large Scale Aptamer-Based Proteomic Profiling to "Planned" Myocardial Infarctions. Circulation. ISSN 1524-4539; 0009-7322

Abstract

Background—Emerging proteomic technologies using novel affinity-based reagents allow for
efficient multiplexing with high sample throughput. To identify early biomarkers of myocardial
injury, we recently applied an aptamer-based proteomic profiling platform that measures 1,129
proteins to samples from patients undergoing septal alcohol ablation for hypertrophic
cardiomyopathy, a human model of “planned myocardial injury” (PMI). Here we examined the
scalability of this approach using a markedly expanded platform to study a far broader range of
human proteins in the context of myocardial injury.
Methods—We applied a highly multiplexed, expanded proteomic technique that uses single
stranded DNA aptamers to assay 4,783 human proteins (4,137 distinct human gene targets) to
derivation and validation cohorts of PMI, to individuals with spontaneous myocardial infarction
(SMI), and to at-risk controls.
Results—We found 376 target proteins that significantly changed in the blood after PMI in a
derivation cohort (n=20; P < 1.05E-05, one-way repeated measures ANOVA, Bonferronithreshold).
Two hundred forty-seven of these proteins were validated in an independent PMI
cohort (n=15; P < 1.33E-04, one-way repeated measures ANOVA); > 90% were directionally
consistent and reached nominal significance in the validation cohort. Among the validated
proteins that were increased within 1 hour after PMI, 29 were also elevated in patients with
spontaneous myocardial infarction (n=63; P < 6.17E-04). Many of the novel markers identified
in our study are intracellular proteins not previously identified in the peripheral circulation or
have functional roles relevant to myocardial injury. For example, the cardiac LIM protein
cysteine and glycine-rich protein 3 (CSRP3) is thought to mediate cardiac mechanotransduction
and stress responses while the mitochondrial ATP synthase F0 subunit component (ATP5J) is a
vasoactive peptide upon its release from cells. Finally, we performed aptamer-affinity
enrichment coupled with mass spectrometry to technically verify aptamer specify for a subset of
the new biomarkers.
Conclusions—Our results demonstrate the feasibility of large scale aptamer multiplexing at a
level that has not previously been reported and with sample throughput that greatly exceeds other
existing proteomic methods. The expanded aptamer-based proteomic platform provides a unique
opportunity for biomarker and pathway discovery following myocardial injury.

Item Type:	Article
Date Deposited:	02 Jan 2018 00:45
Last Modified:	02 Jan 2018 00:45
URI:	https://oak.novartis.com/id/eprint/33075