Method of evaluating the response of ataxia telangiectasia patients to glucocorticoids treatment
The present invention relates to a novel procedure for evaluating the response of patients affected by Ataxia Talangiectasia (A-T) to glucocorticoids treatment. In particular, the procedure provides a step of qualitative and/or quantitative identification in the blood of said patients of the expression of a mRNA variant of the ATM (Ataxia-Talangiectasia-Mutated) gene produced by non-canonical splicing induced by glucocorticoid (GC). In fact, it was demonstrated that mRNA variant expression is present in the blood of patients who responded positively to treatment with GC.
1. A method for evaluating glucocordicoid (GC) treatment response in a patient affected by Ataxia Telangiectasia, comprising
a) amplifying an ATMdexa1 nucleotide sequence in blood from said patient to obtain ATMdexa1 nucleotide expression values with primer pairs:
SEQ ID NO: 1/SEQ ID NO: 33;
SEQ ID NO: 3/SEQ ID NO: 4,
SEQ ID NO: 6/SEQ ID NO: 7,
SEQ ID NO: 9/SEQ ID NO: 34; or
SEQ ID NO: 12/SEQ ID NO: 35,
b) evaluating the ATMdexa1 nucleotide expression values, and
c) administering GC treatment when detectable nucleotide expression values of ATMdexa1 are obtained and reducing and/or discontinuing GC treatment when no detectable expression values of ATMdexa1 are obtained.
2. The method according to claim 1 , wherein the glucocorticoid is prednisolone, dexamethasone, betamethasone, deflazacort, or pharmaceutically acceptable salts or esters thereof or mixtures thereof.
3. The method according to claim 1 , wherein the glucocorticoid is prednisolone phosphate, dexamethasone phosphate, betamethasone phosphate, betamethasone phosphate, deflazacort phosphate or sodium salts thereof.
4. The method according to claim 1 , wherein the ATMdexa1 expression value is normalized versus the expression of a housekeeping gene.
5. The method according to claim 1 , wherein the amplifying is performed by PCR, RT-PCR, RT-PCR with an intercalating agent, Taq polymerase-PCR, Molecular Beacon probe method PCR, FRET-probe hybridization, or Scorpion probe PCR.
6. The method according to claim 5 , wherein the amplifying is carried out using a probe of SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, or SEQ ID NO:15.
7. The method according to claim 1 , wherein the glucocorticoid is a glucocorticoid encapsulated within erythrocytes.
8. The method according to claim 7 , wherein the glucocorticoid is dexamethasone phosphate mono or disodium.
9. A method for treating patients affected by Ataxia Telangiectasia, comprising
a) administering an amount of glucocorticoid to said patient;
b) qualitatively and/or quantitatively identifying expression of ATMdexa1 in blood of said patient, by amplifying said ATMdexa1 nucleic acid with primer pairs:
SEQ ID NO: 1/SEQ ID NO: 33;
SEQ ID NO: 3/SEQ ID NO: 4,
SEQ ID NO: 6/SEQ ID NO: 7,
SEQ ID NO: 9/SEQ ID NO:34; or
SEQ ID NO: 12/SEQ ID NO: 35, and
c) reducing glucocorticoid treatment if there are no detectable expression values for ATMdexa1, or continuing glucocorticoid treatment if there are detectable expression values for ATMdexa1, wherein detectable expression values of ATMdexa1 indicate a positive response to treatment.
10. The method according to claim 9 , wherein the glucocorticoid is prednisolone, dexamethasone, betamethasone, deflazacort or pharmaceutically acceptable salts or esters thereof, preferably phosphate, or mixtures thereof.
11. The method according to claim 9 , wherein the glucocorticoid is a glucocorticoid encapsulated within erythrocytes.
12. The method according to claim 9 , wherein the glucocorticoid is dexamethasone phosphate mono or disodium.
13. The method according to claim 9 , wherein the glucocorticoid is dexamethasone.