Abstract
Background
Transthyretin amyloidosis (ATTR) is a degenerative disease affecting the heart and other organs. Transthyretin (TTR) aggregation is a driver of ATTR pathology, but the mechanism is poorly understood. We used proteomics and tissue clearing technology on wild‐type (WT) human cardiac (WT/WT) and V122I human cardiac (V122I/WT) tissue to better understand TTR cardiomyopathy.
Methods
Flash‐frozen cardiac tissue slices from human subjects with end‐stage WT‐TTR cardiomyopathy, end‐stage V122I TTR cardiomyopathy, and an age‐matched control were used. Fibrils and tissue proteomes were extracted and assessed by bottom‐up proteomics. Tissue clearing was performed using a lauryl sulfate–based lipid removal strategy. Slices were stained using indirect immunofluorescence against targets identified by proteomics. TTR deposits were imaged by antibody and AmyTracker 480 staining. Structures of ATTR fibrils were characterized using cryogenic electron microscopy.