Methods for determining whether a patient suffering from rhabdomyolysis achieves a response with a TLR9 antagonist

Abstract

The inventors initially participated to the identification of LPIN1 mutations as a cause for massive rhabdomyolysis episodes in children, triggered by febrile illness. The inventors have suggested that TLR9 antagonists would be suitable for the treatment of rhabdomyolysis (WO2017085115). The inventors thus treated 2 patients with lipin-1 disease by a TRL9 antagonist (hydroxychloroquine). They showed that the accumulation of mtDNA in plasma of the two patients before treatment decreases under treatment. When the treatment was stopped, the accumulation of mtDNA reappeared, then normalized when treatment was resumed. Accordingly, the present invention relates to a method for determining whether a patient suffering from rhabdomyolysis achieves a response with a TLR9 antagonist comprising determining the amount of mitochondrial DNA (mtDNA) in a blood sample obtained from the patient (e.g. by PCR).

Claims

1. A method for determining whether a patient suffering from rhabdomyolysis achieves a response with a TLR9 antagonist and treating the patient, comprising: (i) determining the amount of mitochondrial DNA (mtDNA) in a blood sample obtained from the patient; (ii) comparing the amount determined at step (i) with a predetermined reference value; and (iii) concluding that the patient achieves a response when the amount determined at step (i) is lower that the predetermined reference value, treating the patient with the TLR9 antagonist.

2. A method of claim for monitoring compliance of a patient being treated for rhabdomyolysis with a TLR9 antagonist, comprising: (i) determining the amount of mitochondrial DNA (mtDNA) in a blood sample obtained from the patient; (ii) comparing the amount determined at step (i) with a predetermined reference value obtained from the patient after treatment began; and (iii) treating the patient with the TLR9 antagonist when an increase in the mtDNA compared to the predetermined reference value is detected, wherein the increase shows that the patient has failed to comply with the prescribed treatment.

3. The method of claim 1, wherein the TFRL9 TLR9 antagonist is hydroxychloroquine.

4. The method of claim 1, wherein the blood sample is a plasma sample.

5. The method of claim 1, wherein the quantification of mtDNA is performed by immunofluorescence or by PCR.

Description

FIGURES

(1) FIG. 1. Inflammatory profile in serum of lipin-1 deficient patients in vivo. Above: IL-6 secretion in serum was measured by ELISA. IL-6 concentration decreases under treatment by Plaquenil® then increases after interruption of treatment then decreases again when the treatment is resumed.

(2) FIG. 2. Quantity of mitochondrial DNA (qPCR) in plasma of two patients. The accumulation of mtDNA before treatment in plasma decreases under treatment by Plaquenil® then increases after interruption of treatment then decreases again when the treatment is resumed.

EXAMPLE

(3) We treated in 2016 two patients (P1, P2) with lipin-1 disease by hydroxychloroquine sulphate (plaquenil®) and two patients (P3, P4) with dantrolene sodium after obtaining the authorization by the CPP of Paris Descarte University (Comité pour la protection des personnes). Moreover the 16-year-old patient (P5) has been treated for one month by plaquenil® because of his cardiac dysfunction, his fatigability and his sleep disorders. Indeed we think that plaquenil® works well on the two patients treated (P1, P2) and P5's state was considered sufficiently serious to propose it. Plaquenil® was given at a dose of 6 mg/Kg/day into 1 intake (dose usually given in Lupus disease) to P1, P2 and P5.

(4) Both patients 1 and 2 treated in vivo with Plaquenil® normalized their serum inflammatory profile in one month after the start of treatment (FIG. 1). IL-6 secretion increased in plasma when the two patients stopped their treatment (FIG. 1). The accumulation of mtDNA in plasma of the two patients before treatment decreases under treatment by Plaquenil® (FIG. 2). After Plaquenil® was stopped, the accumulation of mtDNA reappeared, then normalized when treatment was resumed. We observe a correlation between variations of IL-6 and mtDNA accumulation levels, and with the compliance of the treatment. Both patients also decreased the level of serum Creatine kinase (CK) and did not have any new episode of rhabdomyolysis during 6 months.

(5) Patient 1 corrected his clinical muscle phenotype (Table 1). Physical maximum capacities were measured using six-minute walk (TC6′), Short-Form life quality by using Questionary (SF-36) to assess QOL for parent and for patient, and a self-assessment pain scale (EVA). For patient 2, the pain disappeared but the walking ability did not improve markedly. After 6 months of treatment, the two patients stopped their treatment for lack of compliance after the death of their grandfather, while their mother had to be absent: they immediately underwent extensive rhabdomyolysis and were hospitalized in intensive care (CK 160 000 U/L and 300 000 U/L respectively). Since then they have resumed treatment. Throughout the winter 2016-2017 they did not do rhabdomyolysis and did not have muscle pain whereas winter is usually a difficult time for them.

(6) TABLE-US-00001 TABLE 1 Physical maximum capacities of two patients treated by Plaquenil ®. M 6-M 8 interruption ttt; M 16 Patients 1 and 2 M 0 M 2 M 5 M 9: treatment Six-minute walk P1: 450 m P1: 540 m P1: 520 m P1: 539 m P1: 530 test (normal 560-740) P2: 510 m P2: 550 m P2: 595 m P2: 587 m P2: 590 Questionnary P1: 26 P1: 12 P1: 11 P1: 38 P1: 8 (0: very well - P2: 45 P2: 32 P2: 35 P2: 47 P2: 30 92: very bad) PESDQL children Questionnary P1: 30 P1: 22 P1: 12 P1: 31 P1: 20 PESDQL parent P2: 40 P2: 42 P2: 33 P2: 50 P2: 35 (0: very well - 92: very bad) Pain Evaluation (EVA) P1: 24 P1: 16 P1: 0 P1: 4 P1: 0 (4: not painful - P2: 18 P2: 12 P2: 0 P2: 16 P2: 0 40: very painful)

REFERENCES

(7) Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.

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