WO2020037938A1 - Virus adéno-associé recombinant (raav) ou kit le comprenant, et son application - Google Patents
Virus adéno-associé recombinant (raav) ou kit le comprenant, et son application Download PDFInfo
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- WO2020037938A1 WO2020037938A1 PCT/CN2019/070461 CN2019070461W WO2020037938A1 WO 2020037938 A1 WO2020037938 A1 WO 2020037938A1 CN 2019070461 W CN2019070461 W CN 2019070461W WO 2020037938 A1 WO2020037938 A1 WO 2020037938A1
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- associated virus
- adeno
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- fluorescent protein
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
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- G—PHYSICS
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- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
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Definitions
- the invention relates to the field of biological agents, in particular to a recombinant adeno-associated virus or a kit containing the same and application thereof.
- Gene therapy refers to the introduction of foreign normal target genes into target cells to correct or compensate for diseases caused by genetic defects or abnormalities, including gene modification and gene replacement, and the development of gene vectors has played an important role in gene therapy.
- viral vectors are widely used in gene therapy due to their long-term gene expression and high infection efficiency, especially adeno-associated virus vectors with significant advantages.
- Adeno-associated virus is one of the most widely used vectors in gene therapy.
- Recombinant adeno-associated virus recombinant AAV, rAAV
- recombinant adeno-associated virus vector is non-pathogenic, low immunogenic, specific site integration and stable. The expression of the target gene and other characteristics make it stand out among many viruses and become the best among them.
- the technical problem solved by the present invention is to provide a recombinant adeno-associated virus or a kit containing the same and its application in order to overcome the defect that patients have the risk of developing an immune response during the gene therapy of the recombinant adeno-associated virus.
- one of the technical solutions of the present invention is: the use of a recombinant adeno-associated virus or a kit containing the same in the preparation of a diagnostic product, wherein the diagnostic product is used for
- the diagnostic article is used for diagnosing whether or not the subject to be examined is suitable for adeno-associated virus gene therapy.
- the recombinant adeno-associated virus contains a reporter gene.
- the reporter gene is a fluorescent protein, preferably green fluorescent protein (GFP), deep red fluorescent protein, red fluorescent protein, yellow fluorescent protein, or orange fluorescent protein.
- GFP green fluorescent protein
- the green fluorescent protein GFP is an enhanced green fluorescent protein EGFP, Emerald, sfGFP, superfolded GFP or ZsGreen1 coral isolate protein
- the red fluorescent protein is mCherry, mPlum or mStrawberry
- the yellow fluorescent protein is ZsYellow1 MCitrine, mvenus, or YPet
- the orange fluorescent protein is mOrange, TagRFP, mKO, or dTomato.
- recombinant when used as a modification to vectors, such as viruses, and sequences, such as recombinant polynucleotides and polypeptides, means that the vector or sequence has been manipulated in an artificial manner.
- a specific example of a recombinant virus is the insertion of a polynucleotide into the viral genome, which is not normally found in the wild-type viral genome.
- a recombinant adeno-associated virus or a recombinant adeno-associated virus vector is a clone of a heterologous polynucleotide (eg, a reporter gene) into an adeno-associated virus sequence.
- Recombinant is not always used herein with reference to viral vectors such as AAV viruses and vector sequences, as well as polynucleotides and polypeptides, viral vectors such as AAV, vector sequences, and recombinant forms of polynucleotides and polypeptides are expressly included.
- Recombinant adeno-associated viruses are derived from wild-type adeno-associated viruses, and wild-type genes can be removed from viral sequences by using molecular methods and replaced with non-natural nucleic acids such as reporter transgenes.
- a viral vector as a "recombinant” vector, which in the case of AAV may be referred to as "rAAV”.
- rAAV recombinant adeno-associated virus
- rAAV recombinant adeno-associated virus vector
- rAAV recombinant AAV virus
- the gene therapy is recombinant adeno-associated virus gene therapy (the gene therapy is performed by recombinant adeno-associated virus).
- the recombinant adeno-associated virus is an adeno-associated virus vector rAVV2 carrying a GFP tag, which can also be referred to as rAVV2-GFP.
- the recombinant adeno-associated virus vector may also be derived from AAV1, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh74 or Rh10 vectors, or a hybrid of any of the foregoing AAV adeno-associated viruses or Chimera.
- ex vivo serum, the recombinant adeno-associated virus and mammalian cells are co-cultured for 36-60 hours, preferably 48 hours; the mammalian cells are preferably HEK293 cells, and more preferably 293T cells.
- the screening uses flow cytometry analysis or RT-PCR analysis.
- the “moderate expression” described in the present invention means that the relative expression amount is 0.2 ⁇ expression amount ⁇ 0.6 in the quantitative quantitative PCR, and / or the expression amount is 20% ⁇ expression amount ⁇ 40% in the flow cytometry (control group)
- the expression level of the reporter gene is set to 50%).
- the expression "high expression” in the present invention refers to a relative expression amount of 0.6 ⁇ expression amount ⁇ 1 in fluorescent quantitative PCR, and / or a expression amount of 40% ⁇ expression amount ⁇ 50% (control group) in flow cytometry.
- the expression level of the reporter gene is set to 50%).
- low expression means that the relative expression level is less than 0.2 in the quantitative PCR and / or that the expression level is less than 20% in the flow cytometry (the expression level of the reporter gene in the control group is set to 50). %).
- No expression as used in the present invention, i.e., the expression level of the reporter gene is not measured, or is conventionally understood by those skilled in the art.
- the screening criteria are (wherein the expression level of the reporter gene in the control group is set to 50%): when the expression level of the reporter gene is ⁇ 20%, When ⁇ 40%, the patient is less likely to have an immune response during gene therapy, so he can go directly to gene therapy; when the reported gene expression level is ⁇ 20% or not expressed, the patient is likely to have an immune response during gene therapy. Higher, so patients are not suitable for direct gene therapy.
- the screening criteria are: when the relative expression value of the reporter gene is ⁇ 0.2, preferably ⁇ 0.6, because the patient is less likely to develop an immune response during gene therapy, Therefore, it can be directly used for gene therapy; when the relative expression value of the reporter gene is ⁇ 0.2 or not expressed, the patient has a higher chance of developing an immune response during gene therapy, so it is not suitable for direct gene therapy.
- the criteria for screening are: taking the expression level of the reporter gene in the control group as a reference value (that is, setting the expression level of the reporter gene in the control group to 100%),
- the relative expression of the patient's reporter gene ie, the expression of the reporter gene in the test group / the expression of the reporter gene in the control group
- the relative expression of the patient's reporter gene ie, the expression of the reporter gene in the test group / the expression of the reporter gene in the control group
- the relative expression level of the patient's reporter gene ie, the expression level of the test group's report gene / control group's report gene expression level
- the patient's chance of an immune response during gene therapy High so patients are not suitable for direct gene therapy.
- the screening criteria are: using the GFP fluorescence intensity of the control group as a reference value (that is, the GFP fluorescence intensity of the control group is set to 100%).
- the relative GFP fluorescence intensity that is, the GFP fluorescence intensity in the detection group / the GFP fluorescence intensity in the control group
- the relative GFP fluorescence intensity ie, the GFP fluorescence intensity in the test group / the GFP fluorescence intensity in the control group
- the relative GFP fluorescence intensity ie, the GFP fluorescence intensity in the test group / the GFP fluorescence intensity in the control group
- the patient has a higher chance of developing an immune response during gene therapy, so the patient is not suitable for direct gene therapy .
- the gene for gene therapy is a gene related to Leber's hereditary optic neuropathy.
- the Leber's hereditary optic neuropathy-related gene is a NADH dehydrogenase subunit 1, 4 or 6 gene.
- the detection group is the detection system.
- control group is the reference system.
- control group does not contain the serum sample to be tested, and other components are the same as those of the detection group.
- the serum sample to be tested is replaced with an equal volume of medium or PBS buffer, and other components are the same as those of the detection group.
- the content of each component in the control group is the same as the content of each component in the detection group, except that the serum sample to be tested is not included.
- the medium is a DMEM medium or a DMEM high sugar medium.
- a detection system includes:
- the ratio V0 / V1 of the volume V1 of the serum to be tested to the detection system V0 is 10-100, preferably 20-60.
- the content of the recombinant adeno-associated virus is 5000vg-100000vg / ml, preferably 20,000-40000 / ml.
- the adeno-associated virus host cell is a mammalian cell, preferably a HEK293 cell, and more preferably a 293T cell.
- the content of the adeno-associated virus host cells is 100,000-500000 cells / ml, preferably 30,000-400000 cells / ml.
- the detection system further comprises (d) a culture medium.
- the medium is selected from the group consisting of DMEM medium, DMEM high-sugar medium, or a combination thereof.
- the detection system consists of a reporter gene-containing recombinant adeno-associated virus, a test serum sample, an adeno-associated virus host cell, and optionally a culture medium.
- a system combination includes:
- the reference system does not contain the serum sample to be tested, and other components are the same as those of the detection system.
- the serum sample to be tested is replaced with an equal volume of medium or PBS buffer, and other components are the same as those of the detection system.
- the content of other components is the same as the content of each component in the detection system.
- the medium is a DMEM medium or a DMEM high sugar medium.
- the reporter gene expression level of the detection system is C1
- the reporter gene expression level of the reference system is C0.
- the patient is less likely to develop an immune response during gene therapy
- the patient is more likely to have an immune response during gene therapy.
- a kit is provided, and the kit contains:
- a first reagent comprising a first container and a first recombinant adeno-associated virus containing a reporter gene located in the first container;
- a second reagent comprising a second container, and an adeno-associated virus host cell located in the second container;
- a therapeutic agent comprising a third container and a second recombinant adeno-associated virus located in the third container.
- the second recombinant adeno-associated virus is used for gene therapy.
- the second recombinant adeno-associated virus is a gene therapy drug.
- first reagent and the second reagent may be located in the same or different containers.
- first container and the second container are independent or combined into one.
- an evaluation method includes steps:
- ex vivo serum, the recombinant adeno-associated virus and mammalian cells are co-cultured for 36-60 hours, preferably 48 hours; the mammalian cells are preferably HEK293 cells, and more preferably 293T cells.
- the detection uses flow cytometry analysis or RT-PCR analysis.
- the evaluation method further includes steps:
- step (d) comparing the expression level C1 of the reporter gene detected in step (c) with the expression level C0 of the reference gene (control group).
- the patient when the C1 / C0 ratio is ⁇ 0.4, preferably ⁇ 0.8, the patient is less likely to develop an immune response during gene therapy;
- the patient is more likely to have an immune response during gene therapy.
- the reference system does not contain the serum sample to be tested, and other components are the same as those of the detection system.
- the serum sample to be tested is replaced with an equal volume of medium or PBS buffer, and other components are the same as those of the detection system.
- the content of other components is the same as the content of each component in the detection system.
- the medium is a DMEM medium or a DMEM high sugar medium.
- the reagents and raw materials used in the present invention are all commercially available.
- the positive progress effect of the present invention is as follows: the use of a recombinant adeno-associated virus or a kit containing the same for preparing diagnostic products is provided for the first time, wherein the diagnostic product is used for screening the probability of an immune response in the gene therapy of the recombinant adeno-associated virus.
- the invention also establishes a screening standard for the first time, that is, when the screening is analyzed by flow cytometry, the screening standard is (wherein the expression level of the reporter gene in the control group is set to 50%): If the amount is ⁇ 20%, the patient is less likely to develop an immune response; or, when the screening uses RT-PCR analysis, the screening criteria are: when the relative expression value of the reporter gene is ⁇ 0.2, the patient is likely to develop an immune response Lower. This use has been validated in Leber's hereditary optic neuropathy.
- the standards established by the present invention are widely used, have significant effects, and have strong clinical application value.
- rAAV2-GFP (Guangzhou Paizhen Biotechnology Co., Ltd.) and patient serum (long-term storage -80 ° C, short-term storage -20 ° C, 4 degrees thawing), 293T cells, fetal bovine serum (Gibco, Brazil), DMEM high glucose Medium (Gibco, Brazil), double antibody (10,000 units of penicillin streptomycin double antibody solution Gibco), EDTA, PBS, 24-well plate, flow tube, large culture bottle, bus pipette, pipette tip box and pipette Gun (1ml, 5ml, 10ml), sealing film, 50ml centrifuge tube, alcohol lamp, alcohol, 2ml, 4ml EP tube, high temperature sterilization equipment, centrifuge (Hunan Xiangyi TGL-16M), biological safety cabinet (Thermo 1300 series A2), flow cytometer (Beckman Coulter Cyto FLEX).
- 293T cells cultured with different serum (antibody) concentrations were transfected with rAAV2-GFP using the green fluorescent protein as a reporter gene, and the infection efficiency was sorted by RT-PCR or flow cytometry to compare the rAAV2 for each type
- the infection efficiency of the cells, the affinity of the adeno-associated virus to various types of cells, and the theoretical and experimental basis for the use of adeno-associated virus in gene therapy were further explored.
- the method of the present invention can be used to screen patients with a small adeno-associated virus immune response, thereby effectively avoiding the occurrence of immune response of recombinant adeno-associated virus gene therapy.
- HEK 293 cells (293T cells) were transfected with the GFP-tagged adeno-associated virus rAAV2-GFP to package rAAV2 adeno-associated virus; the adeno-associated virus was concentrated and purified; the titer was subsequently determined. Limited customization. The specific steps are as follows: the day before transfection, 293T cells are seeded in a 225cm 2 cell culture flask, the seeding density is 3.0 ⁇ 10 7 cells / mL, the culture medium is DMEM + 10% bovine serum, and set at 37 ° C.
- High temperature sterilization pipette tip (1ml, 5ml, 10ml), EP tube is placed in the lunch box. Use UV disinfection when drying. Generally, it can be stored for 12 hours after sterilization, which is safer and will not cause cell contamination.
- UV disinfection pipette tip, EP tube, gun, alcohol lamp, marker pen, fetal bovine serum, high sugar, double antibody, EDTA, PBS, 24-well plate, large culture bottle, bus pipette, 50ml centrifuge tube, alcohol lamp
- penicillin-streptomycin mixed solution 10,000 units of penicillin and 10,000 ⁇ g of streptomycin per ml, Gibco
- Passage in culture flasks Passage when the cells become round. After aspirating the waste liquid in the culture flask with a 10ml gun, add 5ml PBS, discard, wash three times, add EDTA, 5ml, put in the incubator for 3min, quickly add 20ml of culture medium, and repeatedly blow with a 10ml gun until the cells are uniformly suspended. Go to another culture bottle, and then base the two culture bottles to 20 ml.
- Example 2 After the 293T cells in Example 2 were cultured on the 24-well plate, the cells became full (approximately 2-3 days) and began to be dispensed.
- Dispensing (recombinant adeno-associated virus dilution): 1960 ⁇ l of DMEM high glucose medium was added with rAAV2-GFP and 40 ⁇ l (4 ml EP tube).
- Serum dilution Configure as follows;
- Group 1 60: 100 ⁇ l patient serum + 500 ⁇ l medium + 300 ⁇ l of the above virus dilution;
- Control group 600 ⁇ l medium + 300 ⁇ l of the above-mentioned virus dilution (or 300 ⁇ l PBS buffer + 300 ⁇ l of medium + 300 ⁇ l of the above-mentioned virus dilution).
- the EP tube is sealed at 4 ° C.
- ⁇ -actin-F cct, aga, ag, c, gt (SEQ ID No .: 1)
- ⁇ -actin-R gag cta cga gcc gtga (SEQ ID NO .: 2)
- GFP-F aca agt tca gcg tgt ccc (SEQ ID NO .: 3)
- GFP-R ctc ggt ggt cgttt gct (SEQ ID NO.:4)
- the total RNA of the cells obtained in Example 3 and Step 5 was extracted using a TRIZOL kit, and a cDNA template was synthesized by reverse transcription.
- Quantitative PCR was performed on a Real-time PCR Detection System instrument.
- a 0.2 mL PCR reaction tube 12.5 ⁇ L of SYBR Green mix, 8 ⁇ L of ddH 2 O, 1 ⁇ L of each pair of primers, 2.5 ⁇ L of cDNA sample, and 25 ⁇ L of the total system were added.
- Each sample was used to amplify both the target gene and the internal reference gene rabbit-actin.
- Each gene was amplified in three replicates.
- the reagents common in each PCR reaction tube can be added together and then aliquoted.
- the fluorescent quantitative PCR was performed according to the pre-denaturation at 95 ° C for 1 s, the denaturation at 94 ° C for 15 s, the annealing at 55 ° C for 15 s, and the extension at 72 ° C for 45 s. A total of 40 cycles of the reaction program were used for amplification. Fluorescent signal. After the reaction, a melting curve analysis of 94 ° C to 55 ° C was performed.
- the relative expression value GFP expression level / ⁇ -actin
- the GFP gene expression level is consistent with the internal control gene expression level as 1.
- GFP expression screening criteria high expression is 0.6 ⁇ relative expression value ⁇ 1, medium expression is 0.2 ⁇ relative expression value ⁇ 0.6, and low expression is less than 0.2.
- High and moderate expression of GFP indicates that the patient's humor is not immune to rAAV2, or the immune response is relatively small, and gene therapy can be performed directly.
- the low expression of GFP indicates that gene therapy using rAAV may cause an immune response, and immunotherapy is required before gene therapy.
- Flow cytometry detection a blank tube needs to be set up for the first time on the machine, that is, a cell group that has not undergone the transfection process is prepared without adding the above drugs, and the same is transferred to the flow tube on the machine, the purpose is to see the GFP of the cell itself Basis value of expression.
- GFP expression level selection criteria control group infection efficiency is 50%: high expression is 40% ⁇ expression level ⁇ 50%, intermediate expression is 20% ⁇ expression level ⁇ 40%, and low expression is below 20%. High and moderate expression of GFP indicates that the patient's humor is not immune to rAAV2 and can be directly used for gene therapy. Low expression of GFP indicates that gene therapy using rAAV may cause an immune response and requires immunotherapy before gene therapy.
- the GFP expression levels detected by patients A, B, C, and D are all higher than 20% (as shown in Table 2-5), which indicates that the patient's humor is not immune to rAAV2, or the immune response that occurs is small and can For gene therapy, especially the GFP expression in D patients is higher than 40%, indicating that the patients are in an ideal state for gene therapy.
- the expression level of GFP detected by patient E is less than 20% (as shown in Table 6), which indicates that gene therapy using rAAV2 may cause an immune response and requires immunotherapy before gene therapy.
- the inventors unexpectedly found that subjects with an expression level of GFP below 20% elicited an immune response during gene therapy. For safety reasons, in this example, no further clinical verification was included. And in the clinic, it is rare to find subjects with expression levels greater than 50%, so the purpose of this embodiment is to verify subjects with expression levels between 20% and 50%. In the clinic, the inventors found that in the 20-50% expression level range, the gene therapy effect has a certain relationship with the expression level, and the subject with an expression level below 40% has a gene therapy effect slightly lower than the expression level at 40 % And above subjects. Therefore, the above low expression, medium expression, and high expression standards were formulated (wherein the GFP expression amount of the control group was set to 50%), and the following verification was performed.
- the subjects Before treatment, screen the subjects for immune response to recombinant adeno-associated virus gene therapy.
- the subjects are grouped according to the amount of GFP expression as follows: high expression is 40% ⁇ expression level ⁇ 50%, that is, group A, intermediate expression is 20% ⁇ expression level ⁇ 40%, that is, group B, in which the GFP expression level of the control group was set to 50%.
- the subjects in this study met the diagnostic criteria for Leber's hereditary optic neuropathy and were identified by gene sequencing as a 11778 site mutation, that is, the arginine of NADH dehydrogenase subunit 4 protein was converted to histidine, which caused dysfunction and optic nerve damage. And Leber hereditary optic neuropathy, which has a high incidence and a poor prognosis (see Chinese Patent CN 102634527 B). Subjects are 10-65 years old, regardless of gender. The clinical-grade rAAV2-ND4 dosage form (State Key Laboratory of Biotherapy of Sichuan University) is used as an injection at a dose of 1 ⁇ 10 10 vg / 0.05ml.
- the route of administration is Intracavity injection, intravitreal injection of 0.05ml under local anesthesia, single administration, clinical observation for 3 months, observation of efficacy, international guidelines for improving vision show that vision improvement of 0.3logMAR (15 letters) is significantly improved, increased by 0.2 logMAR (10 letters) is improved. The increase is less than 0.1logMAR (5 letters). Visual improvement of 0.2 logMAR is effective.
- the method of Embodiment 5 is adopted.
- the experimental result is the average of the three test results (that is, the average value of the GFP fluorescence intensity), and the following formula (I) or (II) is used to calculate the relative GFP fluorescence intensity of the first test group and the second test group;
- Relative GFP fluorescence intensity 1 (First group of GFP fluorescence intensity / Control group GFP fluorescence intensity) ⁇ 100% (I)
- Relative GFP fluorescence intensity 2 (Second group GFP fluorescence intensity / Control group GFP fluorescence intensity) ⁇ 100% ( II)
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Abstract
L'invention concerne une application d'AAV recombinant (rAAV) ou un kit le comprenant dans la préparation d'un produit de diagnostic. Le produit de diagnostic est utilisé pour le criblage pour obtenir la probabilité de réponse immunitaire dans une thérapie génique d'AAV recombinant. L'invention peut déterminer efficacement la probabilité de réponse immunitaire se produisant chez un patient.
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CN202110786630.6A CN113476484A (zh) | 2018-06-29 | 2019-07-01 | 治疗遗传性视神经病变的组合物和方法 |
EP19826653.8A EP3814492A4 (fr) | 2018-06-29 | 2019-07-01 | Compositions et méthodes de traitement de la neuropathie optique héréditaire de leber |
CN201980003485.0A CN110876269B (zh) | 2018-06-29 | 2019-07-01 | 治疗遗传性视神经病变的组合物和方法 |
CN202110786772.2A CN113528510A (zh) | 2018-06-29 | 2019-07-01 | 治疗遗传性视神经病变的组合物和方法 |
SG11202012044QA SG11202012044QA (en) | 2018-06-29 | 2019-07-01 | Compositions and methods for treating leber's hereditary optic neuropathy |
PCT/CN2019/094136 WO2020001657A1 (fr) | 2018-06-29 | 2019-07-01 | Compositions et méthodes de traitement de la neuropathie optique héréditaire de leber |
BR112020026361-3A BR112020026361A2 (pt) | 2018-06-29 | 2019-07-01 | Composições e métodos para tratar neuropatia óptica hereditária de leber |
KR1020217001385A KR102627561B1 (ko) | 2018-06-29 | 2019-07-01 | 레버 유전성 시신경병증의 치료를 위한 조성물 및 방법 |
JP2021521870A JP2021529001A (ja) | 2018-06-29 | 2019-07-01 | レーベル遺伝性視神経症を治療するための組成物及び方法 |
CA3103740A CA3103740A1 (fr) | 2018-06-29 | 2019-07-01 | Compositions et methodes de traitement de la neuropathie optique hereditaire de leber |
AU2019296451A AU2019296451B2 (en) | 2018-06-29 | 2019-07-01 | Compositions and methods for treating leber's hereditary optic neuropathy |
KR1020247001775A KR20240014102A (ko) | 2018-06-29 | 2019-07-01 | 레버 유전성 시신경병증의 치료를 위한 조성물 및 방법 |
MX2020013772A MX2020013772A (es) | 2018-06-29 | 2019-07-01 | Composiciones y métodos para el tratamiento de la neuropatía óptica hereditaria de leber. |
PCT/CN2019/101538 WO2020038352A1 (fr) | 2018-08-20 | 2019-08-20 | Compositions et méthodes de traitement de la neuropathie optique héréditaire de leber |
SG11202101032VA SG11202101032VA (en) | 2018-08-20 | 2019-08-20 | Compositions and methods for treating leber's hereditary optic neuropathy |
KR1020217007727A KR20210068014A (ko) | 2018-08-20 | 2019-08-20 | 레버 유전성 시신경병증의 치료를 위한 조성물 및 방법 |
CN201980054770.5A CN112584874A (zh) | 2018-08-20 | 2019-08-20 | 用于治疗莱伯氏遗传性视神经病变的组合物和方法 |
JP2021509893A JP7403852B2 (ja) | 2018-08-20 | 2019-08-20 | レーベル遺伝性視神経症を治療するための組成物及び方法 |
CA3109432A CA3109432A1 (fr) | 2018-08-20 | 2019-08-20 | Compositions et methodes de traitement de la neuropathie optique hereditaire de leber |
EP19853225.1A EP3840785A4 (fr) | 2018-08-20 | 2019-08-20 | Compositions et méthodes de traitement de la neuropathie optique héréditaire de leber |
AU2019323434A AU2019323434A1 (en) | 2018-08-20 | 2019-08-20 | Compositions and methods for treating leber's hereditary optic neuropathy |
US16/836,644 US11034954B2 (en) | 2018-06-29 | 2020-03-31 | Compositions and methods for treating leber's hereditary optic neuropathy |
US17/181,849 US11352645B2 (en) | 2018-08-20 | 2021-02-22 | Compositions and methods for treating Leber's hereditary optic neuropathy |
US17/317,295 US20220340895A1 (en) | 2018-06-29 | 2021-05-11 | Compositions and methods for treating leber's hereditary optic neuropathy |
US17/320,388 US11332741B1 (en) | 2018-06-29 | 2021-05-14 | Compositions and methods for treating leber's hereditary optic neuropathy |
AU2021204690A AU2021204690A1 (en) | 2018-06-29 | 2021-07-05 | Compositions and methods for treating Leber's hereditary optic neuropathy |
US17/726,833 US20220259619A1 (en) | 2018-08-20 | 2022-04-22 | Compositions and methods for treating leber's hereditary optic neuropathy |
JP2023029170A JP2023078173A (ja) | 2018-06-29 | 2023-02-28 | レーベル遺伝性視神経症を治療するための組成物及び方法 |
JP2023205807A JP2024028861A (ja) | 2018-08-20 | 2023-12-06 | レーベル遺伝性視神経症を治療するための組成物及び方法 |
AU2023285773A AU2023285773A1 (en) | 2018-06-29 | 2023-12-20 | Compositions and methods for treating Leber's hereditary optic neuropathy |
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US11034954B2 (en) | 2018-06-29 | 2021-06-15 | Wuhan Neurophth Biological Technology Limited Company | Compositions and methods for treating leber's hereditary optic neuropathy |
US11352645B2 (en) | 2018-08-20 | 2022-06-07 | Wuhan Neurophth Biotechnology Limited Company | Compositions and methods for treating Leber's hereditary optic neuropathy |
US11357869B2 (en) | 2019-12-09 | 2022-06-14 | Wuhan Neurophth Biotechnology Limited Company | Compositions and methods for treating leber's hereditary optic neuropathy with NADH dehydrogenase proteins |
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US11332741B1 (en) | 2018-06-29 | 2022-05-17 | Wuhan Neurophth Biotechnology Limited Company | Compositions and methods for treating leber's hereditary optic neuropathy |
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US11357869B2 (en) | 2019-12-09 | 2022-06-14 | Wuhan Neurophth Biotechnology Limited Company | Compositions and methods for treating leber's hereditary optic neuropathy with NADH dehydrogenase proteins |
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