Methods for the classification and diagnosis of scoliosis through the use of gi protein receptor

Abstract

Methods for diagnosing a predisposition to developing a scoliosis (e.g., adolescent idiopathic scoliosis (AIS)) and identifying compounds for treating scoliosis based on the modulation of Gi protein-coupled receptor activity are described. Specific embodiments of the methods involve measuring a change in impedance signals of cells expressing a receptor coupled to a Gi protein with a ligand. To identify compounds useful in treatment, the cell is contacted with a test compound and a ligand. A higher impedance in the presence relative to the absence of said test compound is indicative that the test compound is useful for treating scoliosis.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority, under 35 U.S.C. §119(e), of U.S. provisional application Ser. No. 61 / 104,442 filed on Oct. 10, 2008 and of U.S. provisional application Ser. No. 61 / 228,769, filed on Jul. 27, 2009, which are incorporated herein by reference in their entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]N / A.FIELD OF THE INVENTION[0003]The present invention relates to the diagnosis of a predisposition to developing a scoliosis (e.g., adolescent idiopathic scoliosis (AIS)) and to screening assays for identifying compounds for treating scoliosis.BACKGROUND OF THE INVENTION[0004]Scoliosis is a medical condition in which a person's spine is curved from side to side, and may also be rotated. It is an abnormal lateral curvature of the spine. On an x-ray, the spine of an individual with a typical scoliosis may look more like an “S” or a “C” than a straight line.[0005]Spinal deformities and scoliosis in part...

AI Technical Summary

Benefits of technology

[0023]As such, the method of the present invention may also be advantageously used to determine, for example, the type and / or severity of the disease or the nature of the defect underlying the disease (i.e., “classification” or “stratification” of the subjects). This is particularly interesting if the most effective drug for treating or preventing idiopathic scoliosis (e.g., AIS) varies between subjects affected by idiopathic scoliosis of different types and / or severities. The method for classifying of the present invention may therefore permit a better selection of the drug to be used for a particular patient. This method is also useful to classify / stratify already diagnosed scoliotic patients (e.g., adult scoliosis).

[0028]Any sample (e.g., cells, tissues) in which one or more Gi-coupled receptors and / or Gi proteins may be expressed may be used in accordance with the methods of the present invention. The cells may naturally or recombinantly express one or more Gi-coupled receptor(s), and / or a Gi protein. For recombinant expression, a nucleic acid encoding a GiPCR and / or a nucleic acid encoding a Gi protein is introduced into a suitable cell, and the cell is incubated under conditions that provide for expression of the protein(s). The cells used herein naturally express one or more receptors coupled to Gi proteins and were selected in part for their accessibility for collection from subjects. Hence, cells such as osteoblasts, osteoclasts, peripheral blood mononuclear cell (PBMC) (inherently including principally lymphocytes but also monocytes) and myoblasts are advantageously accessible and may conveniently be used in the methods of the present invention. Blood cells (e.g., PBMCs, platelets (thrombocytes), etc.) in particular are particularly accessible and provide for a more rapid testing. Any blood cell can be used for the methods of the present invention so long as it possesses at least one GPCR receptor coupled to a Gi protein. In an embodiment, the cells are obtained or derived from a subject having an idiopathic scoliosis (e.g., Infantile Idiopathic Scoliosis, Juvenile Idiopathic Scoliosis or Adolescent Idiopathic Scoliosis (AIS)). For the present invention, PBMCs do not need to be established in cultures since methods described herein can be performed in cell suspension with fresh or frozen PBMCs.

Problems solved by technology

There is unfortunately no method approved by the FDA yet to identify children or adolescents at risk of developing IS to predict which affected individuals require treatment to prevent or stop progression of the disease (Weinstein S L, Dolan L A, Cheng J C et al.

Consequently, the application of current treatments, such as bracing or surgical correction, is delayed until the detection of a significant deformity or a demonstration of clear progression, resulting in a delayed and less-than-optimal treatment.

About 29,000 scoliosis surgeries are done every year in North America, resulting in significant psychological and physical morbidity.

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