Kit and method for the diagnosis/prognosis of idiopathic scoliosis
Inventors
GARCIA GIMENEZ, JOSE LUIS • BAS HERMIDA, Teresa • PALLARDO CALATAYUD, Federico Vicente • HERVAS MARIN, David • MENA MOLLA, Salvador
Assignees
Hospital Universitario Y Politecnico La Fe • Hospital Universitario Y Politecnico La Fe • Universitat de Valencia • Centro de Investigacion Biomedica en Red CIBER
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Publication Number
US-10975435-B2
Publication Date
2021-04-13
Expiration Date
Abstract
The present invention provides a method for diagnosing or detecting Idiopathic Scoliosis, in particular AIS, as well as a method for prognosticating this disease in a human subject.
Core Innovation
The document describes diagnosing and prognosticating Adolescent Idiopathic Scoliosis (AIS) using circulating plasma microRNAs (miRNAs). It reports a prospective cohort study in which circulating miRNAs are discovered and validated by comparing AIS subjects to healthy controls. The approach uses RT-qPCR to evaluate specific miRNAs in plasma samples and to form a diagnostic and prognostic decision based on measured expression levels relative to control plasma.
A multivariate model computes a probability of AIS from RT-qPCR expression levels of hsa-miR-1306-3p, hsa-miR-223-5p, hsa-miR-27a-5p, and hsa-miR-122-5p. The probability is determined according to Equation 1, and the subject is identified as a human subject in whom AIS is probable. The document also reports model support by diagnostic performance metrics such as ROC curve and AUC, together with additional model criteria.
The document further specifies that treatment of the human subject identified as having probable AIS is performed using one or more of braces; surgery; physical exercise; heparin; warfarin; cyclosporine; glucocorticoids; medroxyprogesterone acetate; thiazide diuretics; melatonin; and a bone morphogenetic protein. Dependent claim refinements include use of X-ray radiographs for imaging conditions relative to the determining and treating steps, and RT-qPCR detection details using forward primer nucleotide sequences referenced by SEQ ID NOs.
Claims Coverage
The independent claim provides a complete method framework that combines RT-qPCR measurement of four circulating plasma miRNAs, computation of an AIS probability using Equation 1, identification of a subject with probable AIS, and subsequent treatment using one or more enumerated AIS therapies. Dependent claims further refine the treatment regimen and add imaging and RT-qPCR primer specificity for particular miRNA detection.
RT-qPCR detection of plasma miRNA panel relative to control
Detecting, by RT-qPCR, an expression level in a test plasma sample from the human subject of each of hsa-miR-1306-3p; hsa-miR-223-5p; hsa-miR-27a-5p; and hsa-miR-122-5p relative, respectively, to the expression level of each of hsa-miR-1306-3p; hsa-miR-223-5p; hsa-miR-27a-5p; and hsa-miR-122-5p in a control plasma sample from a healthy subject.
Computing probability of AIS using Equation 1 from up-regulated miRNAs
Determining, from the expression levels of hsa-miR-1306-3p; hsa-miR-223-5p; hsa-miR-27a-5p and hsa-miR-122-5p that are each up-regulated in the test plasma sample relative, respectively, to the expression levels of hsa-miR-1306-3p; hsa-miR-223-5p; hsa-miR-27a-5p; and hsa-miR-122-5p in the control plasma sample, a probability of AIS in the human subject at risk of AIS according to Equation 1.
Identifying probable AIS subject and treating using enumerated interventions
Identifying the human subject as a human subject in whom AIS is probable; and treating the human subject using one or more of braces; surgery; physical exercise; heparin; warfarin; cyclosporine; glucocorticoids; medroxyprogesterone acetate; thiazide diuretics; melatonin; and a bone morphogenetic protein.
Treatment with braces and/or surgery for probable AIS
Treating the human subject in whom AIS is probable using one or both of braces and surgery.
X-ray radiograph conditions for probable AIS subject
Applying the method where the human subject with probable AIS had previously received an X-ray radiograph.
Exactly one prior X-ray radiograph
Applying the method where the human subject previously received only one X-ray radiograph.
X-ray radiograph between determining and treating
Performing an X-ray radiograph on the human subject where AIS is probable between determining step (ii) and treating step (iii).
RT-qPCR forward primer specificity for hsa-miR-223-5p (SEQ ID NO:13)
Detecting the expression level of hsa-miR-223-5p using a forward primer having the nucleotide sequence of SEQ ID NO:13.
Overall, the claims cover a method that detects a specific plasma miRNA set by RT-qPCR relative to healthy controls, computes an AIS probability using Equation 1, identifies a probable AIS subject, and treats using an enumerated set of interventions, with dependent claims further specifying treatment components, X-ray radiograph conditions, and RT-qPCR primer sequence details for at least hsa-miR-223-5p.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Diagnosing and prognosticating Adolescent Idiopathic Scoliosis (AIS) using circulating plasma microRNAs (miRNAs), including computation of an AIS probability to identify a subject in whom AIS is probable.
Using the identified probable AIS subject to guide treatment with one or more of braces; surgery; physical exercise; heparin; warfarin; cyclosporine; glucocorticoids; medroxyprogesterone acetate; thiazide diuretics; melatonin; and a bone morphogenetic protein.
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