Biomarkers for diagnosis and prognosis of corneal ectatic disorders
The invention relates to the field of the diagnosis and prognosis methods of molecular pathologies. In particular, the invention relates to methods for determining the diagnosis of an ectatic disease of the cornea in a subject, for determining the risk of developing an ectatic disease of the cornea in a subject, for determining the clinical outcome of a subject suffering from an ectatic disease of the cornea and for selecting a subject to be treated with a therapy for an ectatic disease of the cornea based on the determination of the expression levels of TLR2 and/or TLR4 markers. The invention also relates to the use of the TLR2 and/or TLR4 as diagnosis and prognosis markers for an ectatic disease of the cornea.
1. An in vitro method for diagnosing and treating an ectatic disease of the cornea in a subject, or for determining the clinical outcome of a subject suffering from an ectatic disease of the cornea and treating said subject, or for selecting a subject to be treated with a therapy for an ectatic disease of the cornea and treating said subject, said method comprising:
a) obtaining a sample from said subject:
b) determining the expression level of TLR2 and/or TLR4 in the sample from said subject;
c) comparing said expression level with a reference value;
d) diagnosing the subject with an ectatic disease of the cornea, or determining that the subject has a negative clinical outcome, or selecting a subject as a candidate to be treated with a therapy for an ectatic disease of the cornea when the expression level of TLR2 and/or TLR4 is higher than the reference value; and
e) administering a therapy for treating an ectatic disease of the cornea to the subject identified in step (d), wherein said therapy is selected from the group consisting of corneal crosslinking, intracorneal rings, and refractive surgery.
2. The in vitro method according to claim 1 wherein said ectatic disease of the cornea is selected from subclinical keratoconus, clinical keratoconus, pellucid marginal degeneration, keratoglobus and ectasia post refractive corneal surgery.
3. The in vitro method according to claim 2 , wherein said ectatic disease of the cornea is subclinical keratoconus, and the reference value corresponds to the expression levels of TLR2 and/or TLR4 measured in a sample from a healthy subject.
4. The in vitro method according to claim 2 , wherein said ectatic disease of the cornea is clinical keratoconus, and the reference value corresponds to the expression levels of TLR2 and/or TLR4 measured in a sample from a subject suffering from subclinical keratoconus.
5. The in vitro method according to claim 2 , wherein said ectatic disease of the cornea is pellucid marginal degeneration, and the reference value corresponds to the expression levels of TLR2 and/or TLR4 measured in a sample from a healthy subject.
6. The in vitro method according to claim 1 , wherein the sample from said subject is selected from conjunctival and corneal tissue.
7. The in vitro method according to claim 1 , wherein said expression level comprises determining the level of mRNA encoded from the TLR2 and/or TLR4 gene or determining the level of the TLR2 and/or TLR4 protein.
8. The in vitro method according to claim 7 , wherein the protein level is determined by a method selected from immunohistochemistry, Western blot, flow cytometry or by ELISA.
9. The in vitro method for diagnosing an ectatic disease of the cornea in a subject according to claim 1 which also comprises determining at least one parameter selected from: diopters, corneal thickness and corneal elevation.
10. The in vitro method according to claim 1 , wherein step (b) performed by contacting said sample with a composition or kit which comprises an antibody, a polypeptide, a primer and/or probe which specifically bonds to TLR2 and/or TLR4.
11. An in vitro method for determining whether a subject suffering from a refractive defect is a candidate for a refractive surgery therapy or for determining the risk that a subject suffers ectasia following refractive surgery and treating said subject with a suitable therapy, said method comprising:
a) obtaining a sample from said subject;
b) determining the expression level of TLR2 and/or TLR4 in a sample from said subject;
c) comparing said expression level with a reference value;
d) determining that said subject is not a candidate to be treated with a refractive surgery for said refractive defect, or determining that said subject shows high risk of suffering ectasia following refractive surgery when the expression level of TLR2 and/or TLR4 is higher than the reference value; and
e) administering a therapy for treating a refractive defect other than refractive surgery to the subject identified in step (d), wherein said therapy is selected from the group consisting of corneal crosslinking and intracorneal rings.
12. The in vitro method according to claim 11 , wherein the sample from said subject is selected from conjunctival and corneal tissue.
13. The in vitro method according to claim 11 , wherein said expression level comprises determining the level of mRNA encoded from the TLR2 and/or TLR4 gene or determining the level of the TLR2 and/or TLR4 protein.
14. The in vitro method according to claim 13 wherein the protein level is determined by a method selected from immunohistochemistry, Western blot, flow cytometry or by ELISA.