A Transformative Structural Proteomics Platform for Early Detection of Misfolding-Driven Cardiovascular Diseases - Project Summary/Abstract Protein misfolding is a central, disease-driving mechanism in numerous HLBS conditions such as transthyretin amyloid cardiomyopathy, light chain amyloidosis, unexplained heart failure, and atherosclerosis, yet it remains largely invisible to current clinical diagnostics. The Covalent Protein Painting (CPP) platform introduces a transformative solution by enabling direct, site-resolved quantification of protein conformational changes in complex biofluids such as plasma. The proposed project will advance the CPP platform and particularly its first application, the Amyfold assay, a first-in-class blood test to detect misfolded transthyretin (TTR), the key pathogenic event in transthyretin amyloid cardiomyopathy (ATTR-CM). Current diagnostic tools for ATTR-CM rely on imaging, echocardiograms, conventional biomarkers for heart muscle damage or biopsy and fail to detect early, pre-amyloidogenic stages—resulting in delayed diagnoses and missed treatment opportunities. Amyfold overcomes this by offering a minimally invasive, mass spectrometry-based structural biomarker assay that has demonstrated >95% sensitivity and specificity in blinded pilot studies. This capability allows for early diagnosis, better disease monitoring, and improved access to therapies that may prevent or even reverse disease progression. To facilitate clinical applicability and commercialization, the goals of this project are to (1) optimize and validate analytical performance and reproducibility of the assay; (2) demonstrate pre- symptomatic detection in at-risk patients to enable earlier diagnosis and shift ATTR-CM from a fatal to a chronic condition; and (3) establish broader applicability of CPP to other HLBS diseases, including unexplained heart failure, by enabling structural subtyping and precision diagnostics. Building on strong preliminary data and a validated prototype, the project leverages expert collaborators in clinical proteomics and cardiology and a newly developed cloud-based analysis pipeline. The platform’s scalability, compatibility with standard mass spectrometry workflows, and demonstrated high sensitivity support its rapid translation into clinical diagnostics. CPP’s broader applicability to other misfolded proteins (e.g., ApoA1, fibrinogen, APP) further positions it as a platform technology for reshaping early diagnostics across HLBS diseases. By enabling direct measurement of structural pathology before irreversible damage occurs, this technology has the potential to fundamentally change how numerous HLBS diseases are diagnosed, monitored, and treated.
