Vincent Albrecht (Martinsried / DE), Vincenth Brennsteiner (Martinsried / DE), Medini Steger (Martinsried / DE), Aneeta Hotwani (Karachi / PK), Javairia Khalid (Karachi / PK), Imran Nisar (Karachi / PK), Margaret Kasaro (Chapel Hill, NC / US; Lusaka / ZM), Humphrey Mwabe (Lusaka / ZM), Dominique Davis (Chapel Hill, NC / US), Joni Price (Chapel Hill, NC / US), Waqassuddin Khan (Karachi / PK), Fyezah Jehan (Karachi / PK), Kristina De Paris (Chapel Hill, NC / US), Jeffrey SAS Stringer (Chapel Hill, NC / US), Johannes Müller-Reif (Martinsried / DE), Matthias Mann (Martinsried / DE)
Objective
Proteomic technology has progressed tremendously in the past years fueling a revival of mass spectrometry (MS)-based plasma proteomics (Bader, PMID 37209816). We have recently developed a highly robust, deep and economic workflow that should enable the analysis of very large cohorts. We are currently applying this to study adverse pregnancy outcomes (APOs) in low- and middle-income countries and identify biomarkers for improved prediction aiding clinical decision-making.
Methods
The Multi-Omics for Mothers and Infants (MOMI) consortium funded by the Bill & Melinda Gates Foundation has collected more than 40,000 longitudinal plasma samples from women in Africa and South Asia, taken at up to three different time points during pregnancy. Using a semi-automated liquid handling robot, we subjected these samples to our plasma profiling workflow, which features an adaption of the recently reported perchloric-acid enrichment technique (Viode, PMID 36989362). Here, high abundant proteins partition more to the precipitate effectively enriching other plasma proteins in the supernatant. Subsequently we heat-denature and digest them using LysC/trypsin. Peptides are measured on an Orbitrap Astral mass spectrometer interfaced with the Evosep One system using IonOpticks columns.
Here, we report on results of three sub-cohorts accounting in total for more than 10,000 samples provided by the Pakistani AMANHI (Alliance-for-Maternal-and-Newborn-Health-Improvement) and the Zambian IPOP (Improving Pregnancy Outcomes with Progesterone) and ZAPPS (Zambian Preterm Birth Prevention Study) cohorts of MOMI.
Results and Conclusion
The AMANHI cohort with 5,500 plasma samples was measured with a throughput of 60 samples-per-day, yielding on average 2,000 protein groups. Our preliminary data suggest that this in-depth dataset reveals statistically significant changes in more than 1,500 proteins between the first and second trimesters. We identified both known and novel pregnancy-related markers, while corroborating the presence of known markers for APOs such as PAPP-A. Potential novel markers of APOs are currently being validated across the different MOMI subcohort in a "rectangular study approach" (Geyer, PMID 28951502). Initial results from the two Zambian cohorts with more than 5,000 plasma samples are being measured with a throughput of 100 samples-per-day yielding on average 2,400 protein groups. Results appear to support findings from the AMANHI cohort, promising novel biomarkers that would allow for improved early detection of APOs in routine clinical diagnostics.