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In Vivo Somatic Cell Gene Transfer of an Engineered Noggin Mutein Prevents BMP4-Induced Heterotopic Ossification
David L. Glaser, MD1; Aris N. Economides, PhD3; Lili Wang, PhD2; Xia Liu, MS3; Robert D. Kimble, PhD3; James P. Fandl, PhD3; James M. Wilson, MD, PhD2; Neil Stahl, PhD3; Frederick S. Kaplan, MD1; Eileen M. Shore, PhD1
1 Department of Orthopaedic Surgery, University of Pennsylvania, 424 Stemmler Hall, 36th and Hamilton Walk, Philadelphia, PA 19104-6081. E-mail address for D.L. Glaser: david.glaser@uphs.upenn.edu. E-mail address for F. S. Kaplan: frederick.kaplan@uphs.upenn.edu
3 Regeneron Pharmaceuticals, 777 Old Saw Mill River Road, Tarrytown, NY 10591. E-mail address for A.N. Economides: aris@regpha.com
2 Medical Genetics Division, Department of Medicine, University of Pennsylvania, 424 Stemmler Hall, 36th and Hamilton Walk, Philadelphia, PA 19104-6081. E-mail address for D.L. Glaser: david.glaser@uphs.upenn.edu. E-mail address for F. S. Kaplan: frederick.kaplan@uphs.upenn.edu
The Journal of Bone & Joint Surgery.  2003; 85:2332-2342 
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Abstract

Background: The formation of the skeleton requires inductive signals that are balanced with their antagonists in a highly regulated negative feedback system. Inappropriate or excessive expression of BMPs (bone morphogenetic proteins) or their antagonists results in genetic disorders affecting the skeleton, such as fibrodysplasia ossificans progressiva. BMP signaling mediated through binding to its receptors is a critical step in the induction of abnormal ossification. Therefore, we hypothesized that engineering more effective inhibitors of this BMP-signaling process may lead to the development of therapies for such conditions.

Methods: BMP4-induced heterotopic ossification was used as a model for testing the ability of the BMP antagonist Noggin to block de novo bone formation, either by local or systemic delivery. Since Noggin naturally acts locally, a Noggin mutein, hNOG?B2, was engineered and was shown to circulate systemically, and its ability to block heterotopic ossification was tested in a mouse model with use of adenovirus-mediated somatic cell gene transfer.

Results: A mouse model of BMP4-induced heterotopic ossification was developed. Local delivery of wild-type NOG inhibited heterotopic ossification, but systemic administration was ineffective. In contrast, systemic delivery of the adenovirus encoding hNOG?B2 resulted in systemic levels that persisted for more than two weeks and were sufficient to block BMP4-induced heterotopic ossification.

Conclusions: BMP4-induced heterotopic ossification can be prevented in vivo either by local delivery of wild-type Noggin or after somatic cell gene transfer of a Noggin mutein, hNOG?B2. Furthermore, the data in the present study provide proof of concept that a naturally occurring factor can be engineered for systemic delivery toward a desirable pharmacological outcome.

Clinical Relevance: Blocking bone formation is clinically relevant to disorders of heterotopic ossification in humans, such as fibrodysplasia ossificans progressiva. Furthermore, development of BMP antagonists as therapeutic agents may provide modalities for the treatment of other pathologic conditions that arise from aberrant expression of BMPs and/or from a lack of their antagonists.

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    These activities have been planned and implemented in accordance with the Essential Areas and policies of the Accreditation Council for Continuing Medical Education (ACCME) through the joint sponsorship of the American Academy of Orthopaedic Surgeons and The Journal of Bone and Joint Surgery, Inc. The American Academy of Orthopaedic Surgeons is accredited by the ACCME to provide continuing medical education for physicians.
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