Codon-optimized FAM132b prevents diet-induced obesity by modulating adrenergic response and insulin action

Abstract

FAM132b, also known as myonectin, has been identified as a myokine produced by exercise. It is a secreted protein precursor that belongs to the adipolin/erythroferrone family, and has hormone activity in circulation to regulate cellular iron homeostasis and lipid metabolism via unknown receptors. Here, adeno-associated viral vectors (AAV9) were engineered to induce overexpression of FAM132b with 2 codon mutations (A136T and P159A). Treatment of mice under high-fat diet feeding with FAM132b gene transfer resulted in marked reductions in body weight, fat depot, adipocytes size, glucose intolerance and insulin resistance. Moreover, FAM132b overproduction reduced glycemic response to epinephrine (EPI) in whole body and increased lipolytic response to EPI in adipose tissues. This adrenergic response of adipose tissue led to the result that gene transfer reduced glycogen utilization and increased fat consumption in skeletal muscle during exercise. FAM132b knockdown by shRNA significantly increased glycemic response to EPI in vivo and reduced adipocytes response to EPI and adipose tissue browning. Structural analysis suggested that FAM132b mutants delivered by AAV9 may form a weak bond with ADRB2, and potentially bind to insulin against insulin receptor by blocking the receptor binding sites on insulin B-chain. Our study underscores the potential of FAM132b gene therapy with codon optimization to treat obesity by modulating adrenergic response and interfering insulin action. SignificanceWe show here that AAV9-mediated expression of FAM132b with A136T and P159A is a safe and effective therapeutic strategy for improving glucose homeostasis. This is the first demonstration of a therapeutic effect on metabolic disorders in mice with FAM132b codon optimization. These therapeutic effects indicate that FAM132b gene transfer with selective codon mutants in vivo might be a valid therapy for diabetes that can be extended to other metabolic disorders.