Poster #021, Sanford Burnham Prebys
Development of a histological model to validate a novel skeletal muscle regulator’s knockdown in vivo
Mentors: Aashna Lamba, PhD Candidate; Alexandre Colas, PhD
Skeletal muscle is one of the most abundant tissues in the human body, comprising approximately 40% of total body mass, and is indispensable for locomotion, metabolic homeostasis, and systemic health. The maintenance of skeletal mass is governed by a dynamic equilibrium between anabolic and catabolic programs, the disruption of which underlies a broad spectrum of debilitating conditions, from sarcopenia and cachexia to Duchenne muscular dystrophy, that collectively affect hundreds of millions of people worldwide and for which disease-modifying therapies remain critically insufficient. Primary ways to target the muscle mass equilibrium program have been the development of anti-myostatin antibodies, soluble ACVR2B decoy receptors, and follistatin gene therapy, all of which increase muscle mass in preclinical models and some clinical trials. However, these approaches have shown limited functional benefit, dose-limiting toxicities, and incomplete suppression of muscle atrophy, highlighting the need to identify upstream regulators that coordinate both anabolic and anti-catabolic pathways. Thus, there is a need to find novel targets that regulate muscle mass. Using a high-throughput screening approach, our lab identified a novel negative regulator of regenerative skeletal muscle growth. Knocking down this regulator has shown a conserved effect on increasing muscle mass in-vitro (mouse and human) and in-vivo (in mouse injury models). As we continue to test its knockdown in-vivo, it has become necessary to develop specialized tools to validate and quantify knockdown efficiency. Here, we present a histological model created through Biodock and trained on IF microscopy images of in vivo mouse tissue sections, with the goal of recognizing puncta that represent the antibody-stained regulator. The model segments these puncta, distinguishing true positives from background or artifacts. Through this model, we aim to quantify in-vivo knockdown efficiency of the novel regulator and expand its utility to other punctate therapeutic targets in our lab.