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Dysregulation of all-trans retinoic acid metabolism contributes to vascular atherogenesis, restenosis, calcification, and heart failure. Therefore, assessment of proteins involved in retinoid metabolism and signaling has gained interest for identifying potential biomarkers and therapeutic targets in cardiovascular disease. However, quantifying these proteins remains challenging due to limitations of antibody-based methods. We developed a targeted proteomics approach using SureQuant\u2122 internal standard-triggered parallel reaction monitoring mass spectrometry to profile these proteins. We designed a panel of 80 stable isotope-labeled (heavy) peptides representing proteins involved in retinoid signaling and metabolism, with sequences applicable to human samples and conserved across multiple species. Survey experiments using directed data-dependent acquisition on the Orbitrap Fusion Lumos mass spectrometer determined precursor and product ion masses for each heavy peptide, which were programmed into the SureQuant method for continuous monitoring. Upon detection of these heavy internal standards, the instrument transitions to a targeted PRM acquisition mode in which repeated high-resolution MS\/MS spectra of both endogenous (light) and heavy peptides are acquired. Using this method, retinoid pathway-associated proteins were quantified to as low as 10 attomoles for selected targets across multiple tissues and developmental stages. Distinct tissue-specific retinoid metabolic networks were identified across lung, liver, retinal cell lines and cardiac tissues. Developmental profiling of mouse and rat hearts revealed remodeling of retinoid pathway proteins from embryonic to postnatal and adult stages, suggesting a functional transition from retinoid-driven cardiac development toward maintenance of retinoid homeostasis in the mature heart.<\/jats:p>","DOI":"10.64898\/2026.07.27.741027","type":"posted-content","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T12:40:16Z","timestamp":1785242416000},"source":"Crossref","is-referenced-by-count":0,"title":["SureQuant\u2122 IS-PRM Enables Cross-Species Targeted Quantification of Retinoid Metabolism and Signaling Proteins in the Heart"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0136-5821","authenticated-orcid":false,"given":"Sogol","family":"Sedighi","sequence":"first","affiliation":[{"name":"Division of Cardiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7502-3587","authenticated-orcid":false,"given":"Robert N.","family":"O\u2019Meally","sequence":"additional","affiliation":[{"name":"Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sunayana Begum","family":"Syed","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6441-2648","authenticated-orcid":false,"given":"Hugo","family":"Amadei","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1084-4050","authenticated-orcid":false,"given":"Biyi","family":"Li","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4924-2778","authenticated-orcid":false,"given":"Chulan","family":"Kwon","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhishek","family":"Vats","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Johns Hopkins Hospital, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7966-1973","authenticated-orcid":false,"given":"Don","family":"Zack","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Johns Hopkins Hospital, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8093-4465","authenticated-orcid":false,"given":"Kenneth B.","family":"Margulies","sequence":"additional","affiliation":[{"name":"Department of Medicine, Cardiovascular Institute, University of Pennsylvania Perelman, Philadelphia, PA, US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5548-4853","authenticated-orcid":false,"given":"Brian O\u2019","family":"Rourke","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8186-0066","authenticated-orcid":false,"given":"Peter M.","family":"Abadir","sequence":"additional","affiliation":[{"name":"Department of Geriatrics, Johns Hopkins School of Medicine, Baltimore, MD"},{"name":"Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, MD USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert N.","family":"Cole","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9290-9590","authenticated-orcid":false,"given":"D. 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