Metalloprotease activation of myostatin, and methods of modulating myostatin activity

Abstract

It has been determined that metalloprotease cleavage of a myostatin pro peptide results in activation of a latent inactive myostatin to an active form. Accordingly, methods of identifying agents that modulate metalloprotease mediated activation of myostatin are provided, as are agents identified using such methods. Also provided are methods of modulating muscle growth in an organism by increasing or decreasing metalloprotease mediated cleavage of a myostatin pro peptide.

Description

[0001] This application claims the benefit of priority under 35 U.S.C. §119(e) of U.S. Ser. No. 60 / 486,863, filed Jul. 10, 2003; U.S. Ser. No. 60 / 439,164, filed Jan. 9, 2003; and U.S. Ser. No. 60 / 411,133, filed Sep. 16, 2002; the entire content of each which is incorporated herein by reference.[0002] This invention was made in part with government support under Grant Nos. HD35887, AR47746, and GM63471 awarded by the National Institutes of Health. The United States government has certain rights in this invention.BACKGROUND OF THE INVENTION [0003] 1. Field of the Invention [0004] The invention relates generally to metalloprotease regulation of myostatin activity, and more specifically to methods of using agonists or antagonists of the BMP-1 / TLD family of metalloproteases to modulate myostatin activity including, for example, to regulate muscle development in an organism, to methods of identifying agonists and antagonists of such metalloproteases, and to agonists and antagonists so ide...

AI Technical Summary

Benefits of technology

[0017] In one embodiment, the agent decreases proteolytic activity of a metalloprotease that cleaves myostatin pro peptide from a latent myostatin complex, thereby reducing or inhibiting myostatin activation below a level of myostatin activation that occurs or would occur in the absence of the agent. Where such an agent is administered to a subject, the agent can result in increased muscle mass or decreased fat content or both in the subject. The subject can be any subject in which myostatin is expressed, particularly a vertebrate organism, for example, animals that are raised as a food source, such as a mammalian species (e.g., an ovine, porcine species, or bovine species), avian species (e.g, chickens or a turkeys), or a piscine species (e.g., salmon, trout, or cod). The subject also can be a human subject, for example, a subject suffering from a muscular disorder (e.g., a dystonia or dystrophy), a subject suffering from wasting disorder (e.g., cachexia), or a subject suffering from clinical obesity or other metabolic disorder such as type II diabetes. In another embodiment, the agent increases proteolytic activity of a metalloprotease that cleaves myostatin pro peptide from a latent myostatin complex, thereby increasing myostatin activation above a level, if any, of myostatin activation that occurs or would occur in the absence of the agent. Where such an agent is administered to a subject, the agent can result in decreased muscle mass or increased fat content or both in the subject.

[0025] An agent that modulates metalloprotease mediated activation of latent myostatin is exemplified herein by a peptide agent. A peptide agent can include, for example, a peptide portion of a myostatin polypeptide, or a derivative of such a peptide portion of myostatin. In one embodiment, a derivative of a peptide portion of myostatin is a peptide that corresponds to a myostatin pro peptide. In one aspect of this embodiment, the derivative is a pro peptide having a mutation of the metalloprotease cleavage site, for example, a substitution, deletion, or insertion of an amino acid at or in sufficient proximity to the cleavage site such that the metalloprotease has increased or decreased cleavage activity with respect to the peptide agent. In another aspect of this embodiment, the derivative of a peptide portion of myostatin is a peptide agent that reduces or inhibits metalloprotease mediated activation of latent myostatin. The agent that modulates metalloprotease mediated activation of latent myostatin can be operatively linked to a second molecule, which facilitates the action or activity of the agent, or increases or decreases the stability of the agent in a particular environment. For example, a peptide agent can be stabilized by operatively linking the peptide agent to a polypeptide such as an Fc domain of an antibody molecule, thereby increasing the half-life of the peptide agent in vivo.

Problems solved by technology

Unfortunately, these proteases also can act on a variety of other proteins and, therefore, agents that target and inhibit these proteases, for example, signal peptidase, likely would have diverse and deleterious effects if administered to a living organism.

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