Mitochondria are double membrane organelles, containing their own genome, acting as cellular powerhouse via oxidative phosphorylation (Osellame et al., 2012).
Mitochondrial diseases can be caused by mutations in the genes of mitochondrial or nuclear DNA (Koopman et al., 2013). Defects may affect subunits of respiratory chain complexes, mitochondrial assembly proteins, mtDNA maintenance and expression, phospholipid composition of the inner mitochondrial membrane or mitochondrial dynamics; the later controlling the organelle's morphology, with fusion leading to the formation of elongated tubules and fission to isolated punctua. Although many of mitochondrial diseases are multisystemic, some appear to be tissue specific such as optic neuropathies.
Dominant optic atrophy (DOA; OMIM: #165500) is associated with mutations in nuclear genes encoding mitochondrial proteins, primarily the OPA1 gene (opticatrophy gene1 (OPA1; OMIM: *605290)).
Dominant optic atrophy (DOA), Kjer type; or Kjer's autosomal dominant optic atrophy, is an autosomally inherited disease that occurs with an estimated disease prevalence of between 1:12,000 and 1:50,000. (Kivlin, J. D., Lovrien, E. W., Bishop, D. T. & Maumenee, I. H. Linkage analysis in dominant optic atrophy. Am. J. Hum. Genet. 35, 1190-1195 (1983), Kjer, B., Eiberg, H., Kjer, P. & Rosenberg, T. Dominant optic atrophy mapped to chromosome 3q region. II. Clinical and epidemiological aspects. Acta Ophthalmol. Scand. 1996 74, 3-7 (1996) and Lyle, W. M. Genetic risks. Waterloo, Ontario, University of Waterloo Press (1990), Amati-Bonneau et al., 2009, Yu-Wai-Man et al., 2011, Lenaers et al., 2012).
Patients with DOA experience progressive and diffuse atrophy of the retinal ganglion cell layer, loss of myelin and fibrillary gliosis along the anterior visual pathways extending to the lateral geniculate body. This disease is affecting the retina, optic nerves, causing a progressive bilateral reduction in visual acuity beginning in childhood and ultimately could result in blindness.
This pathology remains without effective whether curative or preventive treatment to date, partly due to the complex aetiology of the disease and to unpredictable phases of worsening.
A link between DOA and the OPA1 gene was described in international application WO0227022.
Based on identification of OPA1 mutations as one cause of DOA, a method of diagnosis and treatment of DOA was provided in international application WO00227022. In this application, methods of screening for and detection of carriers of a defective OPA1 gene, diagnosis of a defective OPA1 gene, prenatal OPA1 gene defect screening and detection, gene therapy utilising recombinant technologies and drug therapy using the information derived from the OPA1 gene or OPA1 protein, are disclosed.
A molecular diagnosis is therefore provided by the identification of mutation(s) in the OPA1 gene.
Thus, the majority of patients (about 75%) with DOA harbors at least a mutation in the OPA1 gene (Delettre et al., 2000).
280 different OPA1 mutations have been reported to date (http://mitodyn.org), the majority of which results in premature termination codons and lead to haploinsufficiency by the reduction in OPA1 protein levels (Amati-Bonneau et al 2009).
Other genes were found to be linked to the disease (review in Lenaers et al., 2012), including OPA3 (P Reynier et al., 2004) and more recently NR2F1. (Bosch et al., 2014).
There is a considerable inter- and intra-familial variation in visual acuity, and the penetrance may be as low as about 40% (Cohn et al., 2007).
There is also a marked inter- and intra-familial variability in the rate of disease progression, and a significant proportion of patients (50-75%) will experience further worsening of their visual function in later life (Yu-Wai-Man et al., 2010) (Yu-Wai-Man et al., 2010).
Recent studies evidenced a severe multi-systemic disorder associated with particular OPA1 mutations, named “DOA plus” syndrome (OMIM#125250) (Amati-Bonneau 2008, Zeviani 2008 (Yu-Wai-Man et al., 2010). Nevertheless, although syndromal DOA variants show significant phenotypic variability even within the same family, a consistent finding is a worse visual prognosis among this patient subgroup.
These “DOA plus” patients present additional neurological complications, such as ataxia, sensorineural deafness, chronic progressive external ophtalmoplegia (CPEO) and sensory-motor neuropathy and myopathy in adult life.
Considering these multiple variables, it is difficult to predict when and to which extend a patient with a risk of developing DOA will experience a first symptom, whether said patient may experience complications associated with OPA1 mutations and the seriousness of said complications.
The only method available to evaluate the severity of DOA is the ophthalmological examination.
Using funduscopic examination, the main sign of DOA consists in optic nerve pallor, usually bilateral and symmetric on the temporal side, which is observed in about 50% of patients and is global in the other 50%, especially in old or severely affected patients. However, in moderate cases, the optic nerve atrophy may not be visible.
These ophthalmological examinations are not very sensitive and do not allow to evaluate precisely the disease progression and the extent of damages in retina and optic nerves.
These tests do not allow a sensitive and accurate evaluation or prediction of the severity of the disease or to predict a worsening of the pathological condition(s) and/or whether patient may experience additional complications.
Obviously, invasive tests such as retina biopsies can not be used to evaluate the prognosis of DOA and complications can not be predicted using this method.
Moreover, there are no tests available allowing diagnosing/prognosing OPA1-deficit-induced “DOA plus” syndrome.
There is thus a critical and unmet need for providing effective, sensitive and reliable prognostic of DOA or “DOA plus” syndrome and of its complications, in particular of OPA1-deficit-induced dominant optic atrophy or “DOA plus” syndrome and of its complications.
According to one aspect, the present invention provides means of determining whether or not DOA has a risk of developing, a risk to be affected by a complication a means of assessing its seriousness, and ultimately of identifying the most suitable treatment and of preventing the disease in asymptomatic patients. The process according to the invention will ultimately allow preventive treatment to be administered before irremediable damages of optic neurons and extra-ocular tissues or cells.
In particular, they have demonstrated that the level of expression and/or activity of specific factors are predictive of DOA progression and/or severity.
More particularly, they could identify within DOA patients a subgroup of patients that corresponded to patients experiencing a worsening of the pathological condition.
The method according to this invention particularly makes it possible to anticipate and/or determine DOA seriousness, identify the most suitable treatment. This is an essential means in the follow up of patients.
This invention makes up for the drawbacks of the earlier art by offering a new method, process and kit for prognosis of DOA and opens up the possibility of new preventive treatments.
Additionally, the method according to the invention is simple and reproducible, while being less expensive.
The method according to the invention is more specific and much more sensitive than those previously described. The process according to the invention is non invasive and not painful.
The present invention provides Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease.
The present invention provides the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, and/or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease.
An in vitro prognosis according to the invention is a prognosis performed on a sample collected on a patient by a non-invasive method.
The present invention provides the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, and/or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, in particular wherein said NRF2-activated genes products are selected from the group consisting in NRF2, SOD1, SOD2, catalase, GSTP1, NQO1, Glutathione Reductase, Peroxiredoxin 1, Heme oxigenase 1, Thioredoxin reductase 1, Glutamate Cystein Ligase.
Surprisingly, the inventors have shown that subjects suffering DOA displayed an abnormal level and/or activity of factors involved in the oxidative response in fibroblasts or epithelial cells; further, this change in the level and/or activity of factors involved in the oxidative response in fibroblasts is a hallmark signing an increase in sensitivity and early warning sign of a worsening phase of the disease, or of a complication.
According to an embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, said prognosis being done from birth of said patient.
According to another embodiment, the invention relates to nuclear factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of OPA1 deficiency induced diseases and/or complications comprising detecting said NRF2-activated genes products and another marker of said prognosis such as aconitase.
According to an embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis of OPA-1 deficiency induced diseases and/or complications wherein said use comprises detecting said NRF2-activated genes products and another marker of said prognosis such as aconitase.
According to a preferred embodiment, the invention provides Nuclear factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of OPA-1 deficiency induced diseases and/or complications wherein said use comprises detecting said NRF2-activated genes products and another marker of said prognosis such as aconitase.
According to another embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof of a patient having an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the diagnosis and prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof of a patient suspected to have an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, comprising a first step of diagnosis of said OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications by detecting an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, comprising a first step of diagnosis of said OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications by detecting an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, wherein said prognosis is performed on patients having an OPA1 gene- or OPA1 gene product-deficit-induced disease and for which the mutation of OPA1 gene has been identified.
In fact, the prognosis according to the invention is performed on subjects for whom a first step of diagnosis of said OPA1 gene- or OPA1 gene product-deficit has been realized by detecting an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, blood samples or a mixture thereof, comprising a first step of diagnosis of said OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications by detecting an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, said use comprising a first step of diagnosis of said OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications by detecting an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for their use as above defined wherein OPA1-deficit induced disease is an OPA1-deficit induced optic neuropathy, OPA1-deficit induced autosomal dominant optic atrophy (DOA, OMIM#165500), and/or complications associated with OPA1-deficit induced DOA, in particular—severe multi-systemic syndromes, “DOA plus” disorders as external ophthalmoplegia, ataxia, and deafness or glaucoma, in particular Primary Open Angle Glaucoma, myopathy, peripheral neuropathy, neurodegenerative diseases related to the age (Alzheimer, Parkinson).
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein OPA1-deficit induced disease is an OPA1-deficit induced optic neuropathy, OPA1-deficit induced autosomal dominant optic atrophy (DOA, OMIM#165500), and/or complications associated with OPA1-deficit induced DOA, in particular—severe multi-systemic syndromes, “DOA plus” disorders, external ophthalmoplegia, ataxia, myopathy and deafness or glaucoma, in particular Primary Open Angle Glaucoma, peripheral neuropathy, neurodegenerative diseases related to the age (Alzheimer, Parkinson).
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein OPA1-deficit induced disease is autosomal dominant optic atrophy (DOA, OMIM#165500), and/or complications associated with OPA1-deficit induced DOA, in particular—severe multi-systemic syndromes, “DOA plus” disorders, external ophthalmoplegia, ataxia and deafness or glaucoma, in particular Primary Open Angle Glaucoma.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as defined above, the expression and/or activity of which, determined in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof is modulated with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as defined above, the expression and/or activity of which, determined in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, is modulated with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis, the expression and/or activity of which, determined in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof is modulated with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the expression of which is increased or decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis, the expression of which is increased or decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the expression of which is increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the expression of which is decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the activity of which is increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
In a particularly advantageous embodiment, the expression of said marker is not modified as compared to a control, and the activity of the NRF2-activated genes products for their use according the invention, is increased or decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis, the activity of which is increased or decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
Controls according to the invention may be and is not limited to, healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
A control according to the invention may be the same patient at another time (just before a worsening phase, after a worsening phase, during a worsening phase or latence phase).
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the activity of which is decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the expression and activity of which are decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, the expression and activity of which are increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
According to the present invention an increase means a statistically significant increase as compared to a control ranging from 0.2 to 100 fold increase as compared to a control, preferably 0.5 to 50 fold increase, more preferably a 2 to 100, 5 to 100, 10 to 100, 20 to 100, 50 to 100, and even more particularly a 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50 fold increase as compared to a control.
According to the present invention a decrease means a statistically significant decrease as compared to a control ranging from 0.2 to 100 fold decreased as compared to a control, preferably 0.5 to 50 fold decrease, more preferably a 2 to 100, 5 to 100, 10 to 100, 20 to 100, 50 to 100, and even more particularly a 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50 fold decrease as compared to a control.
A control as used herein, means a negative control. Usually a negative control correspond to the situation or condition without treatment, or with a mock treatment, it corresponds to a condition wherein one expects no modulation.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, wherein said NRF2-activated gene product is a detoxifying enzyme or an antioxidant protein.
According to another embodiment, the invention relates to NRF2-activated genes products for their use as above defined, wherein said NRF2-activated genes products are selected from the group consisting in NRF2, SOD1, SOD2, catalase, GSTP1, NQO1, Glutathione Reductase, Peroxiredoxin 1, Heme oxigenase 1, Thioredoxin reductase 1, Glutamate Cystein Ligase.
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis wherein said NRF2-activated genes products are selected from the group consisting in NRF2, SOD1, SOD2, catalase, GSTP1, NQO1, Glutathione Reductase, Peroxiredoxin 1, Heme oxigenase 1, Thioredoxin reductase 1, Glutamate Cystein Ligase.
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, and/or related complications, in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, wherein said NRF2-activated genes products are selected from the group consisting in NRF2, SOD1, SOD2, catalase, GSTP1, NQO1, Glutathione Reductase, Peroxiredoxin 1, Heme oxigenase 1, Thioredoxin reductase 1, Glutamate Cystein Ligase.
In another more advantageous embodiment, the NRF2-activated genes products according to the invention are Superoxide dismutase 1 (SOD1) (SEQ ID No 1), superoxide dismutase 2 (SOD2) (SEQ ID No 2), catalase (CAT) (SEQ ID No 3), glutathione S-transferase pi 1 (GSTP1) (SEQ ID No 4), NAD(P)H dehydrogenase quinone 1 (NQO1) (SEQ ID No 5), glutathione reductase (GSR) (SEQ ID No 6), thioredoxin reductase 1 (TXNRD1) (SEQ ID No 7), Peroxiredoxin (SEQ ID No 8), heme oxygenase (decycling) 1 (HMOX1) (SEQ ID No 9), glutamate-cysteine ligase modifier subunit (GCLM) (SEQ ID No 10), NRF2 (SEQ ID No 11).
In the invention, said expression and/or activity of said NRF2-activated genes products is detected in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof by immunoblotting or by RT-PCR (Reverse transcription polymerase chain reaction).
In a preferred embodiment, a NRF2-activated gene product expression and/or activity is detected by RT-PCR, using primers SEQ ID No 17 and SEQ ID No 18, SEQ ID No 19 and SEQ ID No 20, SEQ ID No 21 and SEQ ID No 22, SEQ ID No 25 and SEQ ID No 26, SEQ ID No 27 and SEQ ID No 28, SEQ ID No 29 and SEQ ID No 30, SEQ ID No 31 and SEQ ID No 32, or SEQ ID No 33 and SEQ ID No 34.
The invention also relates to a human aconitase for its use in the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease or related complications in a biological sample selected from fibroblasts, blood samples or a mixture thereof in a patient affected or suspected to be affected by said disease.
In the invention, said expression and/or activity of said aconitase is detected in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof by immunoblotting or by RT-PCR and by colorimetric assay.
In a preferred embodiment, the invention relates to a method as defined above, wherein the expression of said NRF2-activated genes products expression and/or activity is detected by RT-PCR, in particular by quantitative RT-PCR and more particularly by quantitative RT-PCR using primers of SEQ ID No 17 and SEQ ID No 18, SEQ ID No 19 and SEQ ID No 20, SEQ ID No 21 and SEQ ID No 22, SEQ ID No 25 and SEQ ID No 26, SEQ ID No 27 and SEQ ID No 28, SEQ ID No 29 and SEQ ID No 30, SEQ ID No 31 and SEQ ID No 32, SEQ ID No 33 and SEQ ID No 34.
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein the prognosis is a prognosis of worsening of the disease and/or related complications.
For patients presenting a “strict DOA” disease, meaning a “light” phenotype of DOA disease without neurological complications, the worsening of the disease and/or related complications means that the “strict DOA” disease evolves to a DOA “plus” syndrome and/or to related complications.
For patients presenting a DOA “plus” syndrome which correspond to patients presenting additional neurological complications, the worsening of the disease and/or related complications means that the syndrome and/or related complications are worsened.
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein, in a group of patients having an OPA1 gene- or OPA1 gene product deficit, a subgroup of patients having:                an expression level of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products lower than that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        and an expression level of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products lower than that of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,is identified as having a prognosis of worsening of the disease and/or related complications.        
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein said NRF2-activated genes products is SOD1 and wherein, in a group of patients having an OPA1 gene- or OPA1 gene product deficit, a subgroup of patients having:                an expression level of SOD1 lower than that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        and an expression level of SOD1 lower than that of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,is identified as having a prognosis of worsening of worsening of the disease and/or related complications.        
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein said NRF2-activated genes products is SOD2 and wherein, in a group of patients having an OPA1 gene- or OPA1 gene product deficit, a subgroup of patients having:                an expression level of SOD2 lower than that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        and an expression level of SOD2 lower than that of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,is identified as having a prognosis of worsening of the disease and/or related complications.        
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein said NRF2-activated genes products are SOD1 and SOD2 and wherein, in a group of patients having an OPA1 gene- or OPA1 gene product deficit, a subgroup of patients having                expression levels of SOD1 and SOD2 lower than those of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        and expression levels of SOD1 and SOD2 lower than those of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,is identified as having a prognosis of worsening of the disease and/or related complications.        
According to another embodiment, the invention relates to the use of Nuclear Factor (erythroid-derived 2)-like 2 (NRF2)-activated genes products for the in vitro prognosis, wherein said NRF2-activated genes products are SOD1 and SOD2 and wherein, in a group of patients having an OPA1 gene- or OPA1 gene product deficit and suffering from DOA pathology, a subgroup of patients having                expression levels of SOD1 and SOD2 lower than those of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        and expression levels of SOD1 and SOD2 lower than those of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,is identified as having a prognosis of worsening of the DOA pathology.        
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis wherein said NRF2-activated genes products is NRF2.
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis wherein said NRF2-activated genes products is NRF2, and wherein the total cell expression of which is increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
According to another embodiment, the invention relates to the use of NRF2-activated genes products for the in vitro prognosis wherein said NRF2-activated genes products is NRF2, and wherein the nuclear translocation of which is increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit.
In an advantageous embodiment, the present invention provides an aconitase or a NRF2-activated gene product for its use in the prognosis of an OPA1-deficit-induced disease and/or complication, the expression and/or activity of said aconitase or of said NRF2-activated gene product is increased or decreased as a function of disease progression, and/or complications.
In this embodiment, the expression and/or activity of said aconitase is increased or decreased as compared to the expression and/or activity of said aconitase measured in the same patient during a worsening phase or a latent phase.
In a particular embodiment, the present invention provides an aconitase, or for its use in the prognosis of an OPA1-deficit-induced disease and/or complication the expression and/or activity of said aconitase was increased or decreased as compared to that of healthy volunteers, as a function of disease progression, and/or complications.
In a preferred embodiment, the present invention provides an human aconitase 2 (SEQ ID No 12), or for its use in the prognosis of an OPA1-deficit-induced disease and/or complication the expression and/or activity of said aconitase was increased or decreased as compared to that of healthy volunteers, as a function of disease progression, and/or complications.
The present invention relates to an in vitro method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprising detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, a modulation in the expression and/or the activity of NRF2-activated genes products with respect to those of healthy subjects of the same age having no deficit in the OPA1 gene or OPA1 gene product or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
The in vitro method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease above defined comprises:                (a) detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression and/or the activity of NRF2-activated genes products.        (b) comparing said expression to those of healthy subjects of the same age having no deficit in the OPA1 gene or OPA1 gene product, or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.        (c) deducing from said comparison whether said individual may expect suffering and/or the severity of said OPA1 gene or OPA1 gene product deficit-induced disease or related complication.        
The in vitro method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease above defined comprises:                (a) detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression and/or the activity of NRF2-activated genes products, and without the use of an invasive sample,        (b) comparing said expression to those of healthy subjects of the same age having no deficit in the OPA1 gene or OPA1 gene product, or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done,        (c) deducing from said comparison whether said individual may expect suffering and/or the severity of said OPA1 gene or OPA1 gene product deficit-induced disease or related complication.        
The in vitro method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease above defined comprises:                (a) detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression and/or the activity of NRF2-activated genes products and without the use of an invasive sample, and in particular without the use of retina sample or optic nerve sample,        (b) comparing said expression to those of healthy subjects of the same age having no deficit in the OPA1 gene or OPA1 gene product, or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done,        (c) deducing from said comparison whether said individual may expect suffering and/or the severity of said OPA1 gene or OPA1 gene product deficit-induced disease or related complication.        
According to an embodiment, the invention relates to a method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprising detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient suspected to be affected by said disease, a modulation in the expression and/or the activity of NRF2-activated genes products with respect to those of a healthy subject having no deficit in the OPA1 gene or OPA1 gene product, from birth of said patient or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to an embodiment, the invention relates to a method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprising detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient suspected to be affected by said disease, a modulation in the expression and/or the activity of NRF2-activated genes products and of another marker of prognosis such as aconitase with respect to those of a healthy subject having no deficit in the OPA1 gene or OPA1 gene product, from birth of said patient or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above for the prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications using a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, in a patient having an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to a method as defined above for the diagnosis and prognosis of an OPA1 gene- or OPA1 gene product-deficit-induced disease, or related complications using a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, from a patient suspected to have an OPA1 gene- or OPA1 gene product-deficit,
According to another embodiment, the invention relates to a method as defined above for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprising detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, a modulation in the expression and/or the activity of NRF2-activated genes products with respect to those of healthy subjects of the same age having no deficit in the OPA1 gene or OPA1 gene product or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done, comprising a first step of diagnosis of said OPA1 gene- or OPA1 gene product-deficit-induced disease by detecting an OPA1 gene- or OPA1 gene product-deficit.
According to another embodiment, the invention relates to a method as defined above, wherein said OPA1-deficit induced disease is an OPA1-deficit induced optic neuropathy, particularly OPA1-deficit induced autosomal dominant optic atrophy (DOA, OMIM#165500), and/or complications associated with OPA1-deficit induced DOA, severe multi-systemic syndromes, “DOA plus” disorders, external ophthalmoplegia, ataxia and deafness and/or glaucoma, in particular Primary Open Angle Glaucoma, myopathy, peripheral neuropathy, neurodegenerative diseases related to the age (Alzheimer, Parkinson).
According to another embodiment, the invention relates to a method as defined above, wherein the expression and/or activity of NRF2-activated genes products, determined in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, is modulated with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein the expression and/or activity of NRF2-activated genes products, determined in a biological sample selected from fibroblasts, epithelial cells, epithelial cells, blood samples or a mixture thereof, is modulated with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said expression is increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above wherein said expression is decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said activity is increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said activity is decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said expression and activity are decreased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said expression and activity are increased with respect to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said NRF2-activated gene product is a detoxifying enzyme or an antioxidant protein.
According to another embodiment, the invention relates to a method as defined above, wherein said NRF2-activated genes products are selected from the group consisting in NRF2, SOD1, SOD2, catalase, GSTP1, NQO1, Glutathione Reductase, Peroxiredoxin 1, Heme oxigenase 1, Thioredoxin reductase 1, Glutamate Cystein Ligase.
According to another embodiment, the invention relates to an in vitro method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprising detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, a modulation in the expression and/or the activity of aconitase with respect to those of a healthy subject of the same age having no deficit in the OPA1 gene or OPA1 gene product.
According to another embodiment, the invention relates to a method as defined above, wherein the expression and/or activity of said NRF2-activated genes products or of aconitase is detected in a biological sample by RT-PCR or by immunoblotting.
According to another embodiment, the invention relates to a method as defined above, wherein the expression of said NRF2-activated genes products expression and/or activity is detected by RT-PCR, in particular by quantitative RT-PCR and more particularly by quantitative RT-PCR using primers designed with the following sequence ID: NRF2, SOD1, SOD2, catalase, GSTP1, NQO1, Glutathione Reductase, Peroxiredoxin 1, Heme oxigenase 1, Thioredoxin reductase 1, Glutamate Cystein Ligase.
In a preferred embodiment, the invention relates to a method as defined above, wherein the expression of said NRF2-activated genes products expression and/or activity is detected by RT-PCR, in particular by quantitative RT-PCR and more particularly by quantitative RT-PCR using primers of SEQ ID No 17 and SEQ ID No 18, SEQ ID No 19 and SEQ ID No 20, SEQ ID No 21 and SEQ ID No 22, SEQ ID No 25 and SEQ ID No 26, SEQ ID No 27 and SEQ ID No 28, SEQ ID No 29 and SEQ ID No 30, SEQ ID No 31 and SEQ ID No 32, SEQ ID No 33 and SEQ ID No 34.
According to another embodiment, the invention relates to a method as defined above, wherein said method is a non invasive method.
The invention also relates to an in vitro method for the prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprising detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, a modulation in the expression and/or the activity of aconitase with respect to those of healthy subjects of the same age having no deficit in the OPA1 gene or OPA1 gene product or to that of healthy subjects of the same age having no OPA1 gene- or OPA1 gene product deficit and to the same patient for whom at least one prognosis test has been previously done.
According to another embodiment, the invention relates to a method as defined above, wherein said modulation in the expression or activity of NRF2-activated genes products or of aconitase is a decrease or an increase.
According to another embodiment, the invention relates to a method for preventing or treating OPA1 deficiency induced diseases and/or complications further comprising administering a treatment comprising administering at least one compound selected from:
Glutathione, Vitamin A, Vitamin C, Vitamin E, Vitamin cofactors (Coenzyme Q10 and Coenzyme Q10 analogs), Minerals (Manganese and Iodide) Carotenoid terpenoids, Natural phenols (Flavonoïdes (such as resveratrol)), Phenolic acids and their esters, Other nonflavonoid phenolics (such as curcuminoids), organic antioxidants (Capsaicin, Bilirubin, oxalic acid, phytic acid, N-Acetylcysteine, R-α-Lipoic acid, fat and water soluble Uric acid), ARE inducers (Sulforafane, Nordihydroguaiaretic acid, Diallyl Sulfid, Diallyl disulfid, Diallyl trisulfid, Pterostilbene, D3T (1,2-dithiole-3-thione), CPDT (5,6-dihydro-cyclopento-(c)-1,2-dithiole-(4H)-thione), Oltipraz, Salicylcurcuminoids, BG12, Bardoxolonemethyl), a combination thereof.
The present invention encompasses these compounds for their use in the prevention and or treatments of worsening phases of DOA, wherein said compounds could be administered daily from birth.
The invention also relates to a kit for the prognosis of OPA1 deficiency induced diseases and/or complications comprising at least one means of detection of NRF2-activated genes products and/or aconitase optionally, comprising at least one means of diagnostic of detection of OPA1 mutated or deficient gene or gene product.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expressions of NRF2-activated genes products, and without the use of retina sample or optic nerve sample,        comparing said expressions to those of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit.        
Said healthy subjects who are used as controls are preferably healthy subjects of the same age as said patients.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expressions of NRF2-activated genes products, and without the use of retina sample or optic nerve sample,        comparing said expressions to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of NRF2-activated genes products, and without the use of retina sample or optic nerve sample,        comparing said expressions to those of same patient having OPA1 gene- or OPA1 gene product deficit at an early stage and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit, and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are lower than those of said healthy subjects, the prognosis is the worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit, and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are similar to those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit, and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are upper than those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are lower than those of said healthy subjects, the prognosis is the worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are similar to those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are upper than those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit, and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are lower than those of the same patients at early stage, the prognosis is associated to a worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit, and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are similar to those of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of NRF2-activated genes products of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to NRF2-activated genes products expressions of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit
and when NRF2-activated genes products expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are upper than those of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the patient prognosis of an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of NRF2-activated genes products, and without the use of retina sample or optic nerve sample,        comparing said expression to those of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        comparing said expression to those of patients at the same age having an OPA1 gene- or OPA1 gene product deficit and having the same expression than that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        deducing from comparison of expression of NRF2-activated genes products of said patient                    to the expression of NRF2-activated genes products of patients at the same age having an OPA1 gene- or OPA1 gene product deficit and having the same expression as that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit                        and to the expression of NRF2-activated genes products of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        that whether said patient has                    expression levels of NRF2-activated genes products lower than those of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,            and expression levels of NRF2-activated genes products lower than those of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,said patient may expect suffering and/or the severity of said OPA1 gene or OPA1 gene product deficit-induced disease or related complication.                        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expressions of SOD 1 and SOD2, and without the use of retina sample or optic nerve sample,        comparing said expressions to those of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit.        
Said healthy subjects who are used as controls are preferably healthy subjects of the same age as said patients.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expressions of SOD 1 and SOD2, and without the use of retina sample or optic nerve sample,        comparing said expressions to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of SOD 1 and SOD2, and without the use of retina sample or optic nerve sample,        comparing said expressions to those of same patient having OPA1 gene- or OPA1 gene product deficit at an early stage and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are lower than those of said healthy subjects, the prognosis is the worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are similar to those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to SOD1 and SOD2 expressions.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are upper than those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to SOD1 and SOD2 expressions.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are lower than those of said healthy subjects, the prognosis is the worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are similar to those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to SOD1 and SOD2 expressions.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are upper than those of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to SOD1 and SOD2 expressions.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are lower than those of the same patients at early stage, the prognosis is associated to a worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are similar to those of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to SOD1 and SOD2 expressions.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of SOD 1 and SOD2 of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to SOD1 and SOD2 expressions of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when SOD1 and SOD2 expressions of patient having an OPA1 gene or OPA1 gene product deficit-induced disease are upper than those of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to SOD1 and SOD2 expressions.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of SOD 1 and SOD2, and without the use of retina sample or optic nerve sample,        comparing said expression to those of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        comparing said expression to those of patients at the same age having an OPA1 gene- or OPA1 gene product deficit and having the same expression than that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,        deducing from comparison of expression of SOD1 and SOD2 of said patient                    to the expression of SOD1 and SOD2 of patients at the same age having an OPA1 gene- or OPA1 gene product deficit and having the same expression as that of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit            and to the expression of SOD1 and SOD2 of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,            that whether said patient has                            expression levels of SOD1 and SOD2 lower than those of healthy subjects at the same age having no OPA1 gene- or OPA1 gene product deficit,                and expression levels of SOD1 and SOD2 lower than those of patients at the same age having an OPA1 gene- or OPA1 gene product deficit,said patient may expect suffering and/or the severity of said OPA1 gene or OPA1 gene product deficit-induced disease or related complication.                                                
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of catalase, and without the use of retina sample or optic nerve sample,        comparing said expressions to that of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit.        
Said healthy subjects who are used as controls are preferably healthy subjects of the same age as said patients.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of catalase, and without the use of retina sample or optic nerve sample,        comparing said expressions to that of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the expression of catalase, and without the use of retina sample or optic nerve sample,        comparing said expressions to that of same patient having OPA1 gene- or OPA1 gene product deficit at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is lower than that of said healthy subjects, the prognosis is the worsening of the disease and/or related complications.
Said healthy subjects who are used as controls are preferably healthy subjects of the same age as said patients.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit, and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is similar to that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to catalase expression.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expressions of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is upper than that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to catalase expression.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is lower than that of said healthy subjects, the prognosis is the worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is similar to that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to catalase expression.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is upper than that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to catalase expression.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is lower than that of the same patients at early stage, the prognosis is associated to a worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is similar to that of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to catalase expression.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprising the comparison of the expression of catalase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to catalase expression of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when catalase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is upper than that of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to catalase expression.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the activity of aconitase, and without the use of retina sample or optic nerve sample,        comparing said expressions to that of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit.        
Said healthy subjects who are used as controls are preferably healthy subjects of the same age as said patients.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the activity of aconitase, and without the use of retina sample or optic nerve sample,        comparing said expressions to that of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises:                detecting in a biological sample selected from fibroblasts, epithelial cells, blood samples or a mixture thereof, of a patient affected or suspected to be affected by said disease, the activity of aconitase, and without the use of retina sample or optic nerve sample,        comparing said expressions to that of same patient having OPA1 gene- or OPA1 gene product deficit at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit.        
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase expression of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when aconitase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is lower than that of said healthy subjects, the prognosis is associated to a worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the activity of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase activity of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when aconitase activity of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is similar to that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the activity of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase activity of healthy subjects having no OPA1 gene- or no OPA1 gene-product deficit,
and when aconitase activity of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is upper than that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase expression of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when aconitase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is lower than that of said healthy subjects, the prognosis is associated to a worsening of the disease and/or related complications
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase activity of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when aconitase activity of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is similar to that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the activity of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase expression of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene-product deficit,
and when aconitase activity of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is upper than that of said healthy subjects, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase activity of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when aconitase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is lower than that of the same patients at early stage, the prognosis is associated to a worsening of the disease and/or related complications.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase expression of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when aconitase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is similar to that of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.
According to an embodiment of the invention, the in vitro method for the prognosis of patient having an OPA1 gene or OPA1 gene product deficit-induced disease comprises the comparison of the expression of aconitase of patient having an OPA1 gene or OPA1 gene product deficit-induced disease, to aconitase expression of the same patient at an early stage, and to those of healthy subjects at the same age having no OPA1 gene- or no OPA1 gene product deficit,
and when aconitase expression of patient having an OPA1 gene or OPA1 gene product deficit-induced disease is upper than that of the same patients at early stage, the prognosis is that the disease and/or related complications are not worsened with respect to the antioxidant mechanism related to the NRF2 activation.