Vector production

• US 10,912,824 B2
• Filed: 07/13/2015
• Issued: 02/09/2021
• Est. Priority Date: 07/14/2014
• Status: Active Grant

• Current Assignee: Fondazione Telethon Ets, Ospedale San Raffaele Srl (Policlinico San Donato S.p.A.)
• Original Assignee: Ospedale San Raffaele Srl (Policlinico San Donato S.p.A.)
• Inventors: Cantore, Alessio, Lombardo, Angelo Leone, Naldini, Luigi
• Primary Examiner(s): Horning, Michelle S
• Application Number:
  US15/325,124
• Publication Number:
  US 20170165348A1
• Time in Patent Office: 2,038 Days
• Field of Search: None
• CPC Class Codes: A61K 2121/00 Preparations for use in therapy
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 11/02 Nasal agents, e.g. decongestants
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 15/08 for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/04 Antipruritics
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/06 Antimigraine agents
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/04 : Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 15/86 : Viral vectors
• CPC Class Codes: C12N 2740/10052 : relating to complementing cells and packaging systems for producing virus or viral particles
• CPC Class Codes: C12N 2740/16041 : Use of virus, viral particle or viral elements as a vector
• CPC Class Codes: C12N 2740/16052 : relating to complementing cells and packaging systems for producing virus or viral particles
• CPC Class Codes: C12N 7/04 : Inactivation or attenuation; Producing viral sub-units
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

An enveloped viral particle producer or packaging cell, wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell.

Claims

1. An enveloped viral particle producer or packaging cell, wherein:

the producer or packaging cell is genetically engineered to decrease expression of MHC-I on the surface of the producer or packaging cell such that the number of surface-exposed MHC-I molecules is less than 50% of the number of surface-exposed MHC-I molecules that are displayed in the absence of the genetic engineering, and

wherein the producer or packaging cell comprises one or more nucleic acid sequences encoding one or more viral particle structural proteins; and

wherein;

(a) the packaging cell lacks a vector encoding a viral genome;

whereas

(b) the producer cell further comprises a vector encoding a viral genome comprising a therapeutic nucleotide of interest.

2. The enveloped viral particle producer or packaging cell of claim 1 wherein the cell comprises a genetically engineered disruption of a gene encoding β

2-microglobulin.

3. The enveloped viral particle producer or packaging cell of claim 1 wherein the cell comprises a genetically engineered disruption of one or more genes encoding an MHC-I α

chain.

4. The enveloped viral particle producer or packaging cell of claim 1, wherein the cell is a HEK-293 cell or a derivative thereof.

5. The enveloped viral particle producer or packaging cell of claim 4, wherein the cell is a HEK-293T or a HEK-293 T-REx cell.

6. The enveloped viral particle producer or packaging cell of claim 1, wherein the enveloped viral particle is a retrovirus, herpes simplex virus or vaccinia virus, hepadnavirus, togavirus, flavivirus, arenavirus, coronavirus, orthomyxovirus, paramyxovirus, bunyavirus, bornavirus, rhabdovirus, filovirus or derived therefrom.

7. The enveloped viral particle producer or packaging cell of claim 1, wherein the enveloped viral particle is derived from a retrovirus or herpes simplex virus or vaccinia virus.

8. The enveloped viral particle producer or packaging cell of claim 1, wherein the enveloped viral vector is derived from a lentivirus.

9. The enveloped viral particle producer or packaging cell of claim 1, wherein the cell comprises one or more nucleic acid sequences encoding a viral capsid protein and/or a viral envelope protein.

10. The enveloped viral particle producer or packaging cell of claim 1, wherein the viral particle is a retroviral particle, and wherein the cell comprises one or more nucleic acid sequences encoding one or more proteins selected from the group consisting of Gag, Pol and Env.

11. The enveloped viral particle producer or packaging cell of claim 10, wherein the viral particle is a lentiviral particle.

12. The enveloped viral particle producer or packaging cell of claim 11, wherein the cell further comprises a nucleic acid sequence encoding Rev.

13. The enveloped viral particle producer or packaging cell of claim 1, wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell such that the number of surface-exposed MHC-I molecules is less than 40% of the number of surface-exposed MHC-1 molecules that are displayed in the absence of the genetic engineering.

14. The enveloped viral particle producer or packaging cell of claim 1, wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell such that the number of surface-exposed MHC-I molecules is 0% of the number of surface-exposed MHC-I molecules that are displayed in the absence of the genetic engineering.

15. The enveloped viral particle producer or packaging cell of claim 1, wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell such that the cell is substantially devoid of surface-exposed MHC-I molecules.

16. A method of producing enveloped viral particles comprising the steps of:

a) providing an enveloped viral particle producer cell according to claim 1; and

b) culturing the cell under conditions suitable for the production of the enveloped viral particles.

17. The method of claim 12, wherein the enveloped viral particles are substantially devoid of surface-exposed MHC-1 molecules.

18. An enveloped viral particle derived from a retrovirus, herpes simplex virus, vaccinia virus, hepadnavirus, togavirus, flavivirus, arenavirus, coronavirus, orthomyxovirus, paramyxovirus, bunyavirus, bornavirus, rhabdovirus or filovirus, wherein the particle is produced according to the method of claim 12 and is substantially devoid of surface-exposed MHC-I molecules.

19. A cell transduced by the enveloped viral particle of claim 18.

20. A pharmaceutical composition comprising the enveloped viral particle of claim 18, or comprising a cell transduced by said enveloped viral particle, and a pharmaceutically acceptable carrier, diluent or excipient.

21. A method of gene therapy comprising transducing a cell with the enveloped viral particle of claim 18.

22. The method of claim 21 wherein the transduction is carried out ex vivo.

23. A method of gene therapy comprising administering the enveloped viral particle of claim 18, or a cell transduced by said enveloped viral particle, to a subject in need thereof.

24. A method of vaccination comprising administering the enveloped viral particle of claim 18 to a subject in need thereof.