+

WO1994023582A1 - Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons - Google Patents

Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons Download PDF

Info

Publication number
WO1994023582A1
WO1994023582A1 PCT/US1994/003831 US9403831W WO9423582A1 WO 1994023582 A1 WO1994023582 A1 WO 1994023582A1 US 9403831 W US9403831 W US 9403831W WO 9423582 A1 WO9423582 A1 WO 9423582A1
Authority
WO
WIPO (PCT)
Prior art keywords
adenoviral
vector
surfactant protein
free
lung surfactant
Prior art date
Application number
PCT/US1994/003831
Other languages
English (en)
Other versions
WO1994023582A9 (fr
Inventor
Bruce Trapnell
Jeffrey Whitsett
Original Assignee
Genetic Therapy, Inc.
University Of Cincinnati Children's Hospital
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Genetic Therapy, Inc., University Of Cincinnati Children's Hospital filed Critical Genetic Therapy, Inc.
Priority to JP6523310A priority Critical patent/JPH09500782A/ja
Priority to EP94914075A priority patent/EP0701401A4/fr
Publication of WO1994023582A1 publication Critical patent/WO1994023582A1/fr
Publication of WO1994023582A9 publication Critical patent/WO1994023582A9/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/785—Alveolar surfactant peptides; Pulmonary surfactant peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

  • This invention relates to adenoviral vectors. More particularly, this invention relates to adenoviral vectors which include DNA encoding a lung surfactant protein and to the use of such vectors in treating disease states associated with lung surfactant protein deficiency, such as infant respiratory distress syndrome, and adult respiratory distress syndrome.
  • Surfactant proteins are natural endogenous proteins produced primarily within the alveolar and airway epithelial cells of the normal lung and interact with phospholipids to maintain the patency of the alveolar structures.
  • pulmonary surfactant protein concentration on the alveolar surface falls below critical levels, surface tension of the liquid-gas interface increases, thereby leading to alveolar collapse, pulmonary ventilation-perfusion mismatch, and hypoxia. In severe cases, this can lead to death.
  • Intermittent administration of exogenous bovine lung surfactant protein has shown partial, but not complete remission of the pathophysiology of the surfactant deficiency state.
  • an adenoviral vector including a DNA sequence encoding a lung surfactant protein.
  • the adenoviral vector is a replication deficient adenoviral vector, i.e., such vector is free of a DNA sequence(s) which is (are) required for viral replication, such as, for example, the El DNA sequence or a portion thereof.
  • the adenoviral vector is free of at least a portion of the adenoviral El DNA sequence and is free of at least a portion of the adenoviral E3 DNA sequence.
  • the E3 region encodes several polypeptides which help the adenoviru ⁇ to evade the immune surveillance of the host.
  • the adenoviral vector comprises an adenoviral 5' inverted terminal repeat, or ITR; an adenoviral 3' ITR; an adenoviral encap ⁇ idation signal; the DNA sequence encoding a lung surfactant protein; and a promoter controlling the expression of the DNA sequence encoding the lung surfactant protein.
  • the vector is free of at least the majority of the adenoviral El and E3 DNA sequences, but is not free of all of the E2 and E4 DNA sequences, and is not free of DNA sequences encoding adenoviral proteins expressed by the adenoviral major late promoter.
  • the vector is also free of at least a portion of at least one DNA sequence selected from the group consisting of the E2 and E4 DNA sequences.
  • the vector is free of the adenoviral El and E3 DNA sequences, and is free of one of the E2 and E4 DNA sequences, and is free of a portion of the other of the E2 and E4 DNA sequences.
  • the vector is free of at least the majority of the El and E3 DNA sequences, is free of at least a portion of at least one DNA sequence selected from the group consisting of the E2 and E4 DNA sequences, and is free of DNA sequences encoding adenoviral proteins expressed under control of the adenoviral major late promoter.
  • Suitable promoters which may be employed include, but are not limited to, adenoviral promoters, such as the adenoviral major late promoter; or heterologou ⁇ promoters, such as the cytomegalovirus (CMV) promoter; the Rous Sarcoma Virus promoter; the respiratory syncytial virus promoter; inducible promoters, such as the mouse mammary tumor virus, or MMTV, promoter; the metallothionein promoter; and heat shock promoters.
  • tissue-specific promoters such as, but not limited to, lung surfactant protein promoters, may also be employed. It is to be understood, however, that the scope of the present invention is not to be limited to any specific promoter.
  • Lung surfactant proteins which may be encoded by the DNA sequence encoding a lung surfactant protein include surfactant protein A (SPA), surfactant protein B (SPB), and surfactant protein C (SPC) .
  • SPA surfactant protein A
  • SPB surfactant protein B
  • SPC surfactant protein C
  • Surfactant protein A is described in Kuroki, et al., J. Biol. Chem.. Vol. 263, No. 7, pgs. 3388-3394 (March 5, 1988).
  • Surfactant protein B and DNA encoding therefor are described in Pilot-Matias, et al., DNA / Vol. 8, No. 2, pgs. 75-86 (1989), Glasser, et al., Proc. Nat. Acad. Sci.. Vol 84, pgs. 4007-4011 (June 1987); Revak, et al., J. Clin. Invest.. Vol. 81, pgs.
  • the DNA sequence encoding a lung surfactant protein encodes lung surfactant protein B.
  • a vector in a preferred embodiment, is assembled first by constructing, according to standard techniques, a shuttle plas id which contains, beginning at the leftward adenoviral geno ic elements, the "critical left end elements", which include an adenoviral 5' ITR, an adenoviral encapsidation signal, and the Ela enhancer sequence; a promoter (which may be an adenoviral promoter or a foreign promoter); a tripartite leader sequence, a multiple cloning site; a poly A signal; and a DNA segment which corresponds to a segment of the adenoviral genome.
  • Such DNA segment serves as a substrate for homologous recombination with a modified or mutated adenovirus, and such sequence may encompass, for example, a segment of the adenoviral genome from base 3328 to base 6241 of the adenovirus 5 genome.
  • the plasmid may also include a selectable marker and an origin of replication.
  • the origin of replication may be, for example, a bacterial origin of replication.
  • a representative example of such a shuttle plasmid is pAVS6, shown in Figure 4.
  • An intron may be included within the transcribed portion to enhance the cytoplasmic mRNA accumulation levels.
  • the multiple cloning site facilitates the insertion of the DNA sequence encoding a lung surfactant protein into the plasmid.
  • the DNA sequence encoding the lung surfactant protein may be inserted into the multiple cloning site.
  • restriction enzyme sites separating the above-mentioned components of the shuttle plasmid include "rare" restriction enzyme sites; i.e., sites which are found to occur randomly in eukaryotic genes at a frequency from about one in every 10,000 to about one in every 100,000 base pairs. This increases the flexibility and ease of rearranging components of the vectors in assembled shuttle plasmids.
  • Homologous recombination is then effected with a modified or mutated adenovirus in which at least the majority of the El and E3 adenoviral DNA sequences have been deleted, as shown, for example, in Figure 8.
  • a modified or mutated adenovirus in which at least the majority of the El and E3 adenoviral DNA sequences have been deleted, as shown, for example, in Figure 8.
  • Such homologous recombination may be effected through co-transfection of the shuttle plasmid and the modified adenovirus into a helper cell line, such as 293 (embryonic kidney epithelial) cells, by CaP0 4 precipitation.
  • a cloning vector is formed in which the modified adenovirus DNA which was 5' to the DNA segment in the shuttle plasmid corresponding to a similar segment of the modified adenoviral genome is replaced with the components in the shuttle plasmid which are 5' to such DNA segment.
  • This homologous recombination, or "crossing over" event can occur anywhere along the segment of the genome of the modified adenovirus which corresponds to the segment which is also contained within the shuttle plasmid (such as, for example, bases 3328 to 6241 of adenoviru ⁇ 5 in Example 1 shown below) .
  • a vector which includes an adenoviral 5' ITR; an adenoviral encapsidation signal; an Ela enhancer sequence; a promoter; a tripartite leader sequence; a DNA sequence encoding a lung surfactant protein; a poly A signal; adenoviral DNA free of at least the majority of the El and E3 adenoviral DNA sequences; and an adenoviral 3' ITR.
  • This vector may then be introduced into a cell line such as the 293 cell line for production of large amounts of infectious recombinant adenoviral particles.
  • the 293 cell line is a human fetal kidney epithelial cell line into which has been permanently introduced 11% of the left end of the adenovirus 5 genome. This directs the synthesis of the adenoviral Ela and Elb proteins and allows trans-complementation of El-deleted vectors.
  • infectious viral particles may then be administered to a host in vivo as part of a gene therapy procedure.
  • infectious viral particles may be administered sy ⁇ temically, such as by intravenous or intraperitoneal or intrasmuscular or subcutaneous administration, or may be administered topically, such as by intratracheal or intrabronchial administration, or, alternatively, the infectious viral particles may be administered in an aerosol formulation.
  • the infectious viral particles may be administered in an amount of up to about 10 13 pfu, preferably from about 10 7 pfu to about 10 12 pfu.
  • the infectious viral particles may be employed in the transduction of the epithelium of the respiratory tract or alveoli; whereby the lung epithelial cells will express lung surfactant protein in amounts sufficient to achieve clinical correction of lung surfactant protein deficiency.
  • infectious viral particles may be used to transduce eukaryotic cells in vitro.
  • Eukaryotic cells which may be transduced include, but are not limited to, macrophages, lymphocytes, fibroblasts, liver cells, bronchial cells, and other epithelial or endothelial cells.
  • Such eukaryotic cells then may be administered to a host as part of a gene therapy procedure, or may be cultured in vitro whereby such cells produce lung surfactant protein.
  • the infectious viral particles may be used to transduce eukaryotic cells in vitro for the in vitro production of lung surfactant protein.
  • eukaryotic cells which may be transduced in vitro for the in vitro production of lung surfactant protein include, but are not limited to, those eukaryotic cells hereinabove described, as well as Chinese Hamster Ovary (CHO) cells, COS-7 cells, NIH 3T3 cells, vero cells, HeLa cells, MRC-5 cells, CN1 cells, W138 cells, and chicken lymphoma cells.
  • the lung surfactant protein produced by such cells may then be administered to a host in conjunction with an acceptable pharmaceutical carrier in order to treat lung surfactant protein deficiency states.
  • the vector comprises an adenoviral 5' ITR; an adenoviral 3' ITR; an adenoviral encapsidation signal; a DNA sequence encoding a lung surfactant protein; and a promoter controlling the DNA sequence encoding a lung surfactant protein.
  • the vector is free of the adenoviral El, E2, E3, and E4 DNA sequences, and the vector is free of DNA sequences encoding adenoviral proteins promoted by the adenoviral major late promoter; i.e., the vector is free of DNA encoding adenoviral structual proteins.
  • Such vector is sometimes hereinafter referred to as a "gutless adenoviral vector," or "GLAd" vector.
  • Promoters which are contained in the vector may be those hereinabove described.
  • Such vectors may be constructed by removing the adenoviral 5' ITR, the adenoviral 3' ITR, and the adenoviral encapsidation signal, from an adenoviral genome by standard techniques.
  • Such components, as well as a promoter (which may be an adenoviral promoter or a non-adenoviral promoter), tripartite leader sequence, poly A signal may, by standard techniques, be ligated into a base plasmid or "starter" plasmid such as, for example, pKSII"(Strategene) , to form an appropriate cloning vector.
  • the cloning vector may include a multiple cloning site, as hereinabove de ⁇ cribed, to facilitate the insertion of the foreign DNA sequence into the cloning vector.
  • An appropriate vector in accordance with the present invention is thus formed by cutting the cloning vector by standard techniques at appropriate restriction sites in the multiple cloning site, and then ligating the DNA sequence encoding a lung surfactant protein into the cloning vector.
  • the GLAd vector may then be packaged into infectious viral particles using a helper adenovirus or cell line which provides the necessary packaging materials.
  • a helper virus is used, in one embodiment, preferably it has a defective encapsidation signal in order that the helper virus will not package itself.
  • Examples of such encapsidation-defective helper viruses which may be employed are described in Grable, et al. , J. Virol.. Vol. 66, pgs. 723-731 (1992), and in Grable, et al., J. Virol.. Vol. 64, pgs. 2047-2056 (1990).
  • the helper virus has a normal packaging signal.
  • DNA for the vector and the encapsidation-defective helper virus are transfected into an appropriate cell line for the generation of infectious viral particles. Transfection may take, place by electroporation, calcium phosphate precipitation, microinjection, or through proteoliposomes. Examples of appropriate cell lines include, but are not limited to, HeLa cells, A549 cells, or 293 (embryonic kidney epithelial) cells.
  • the infectious viral particles may then be purified away from helper virus by CsCl isopycnic density centrifugation and transduced into lung epithelial cells lining the respiratory tract or alveoli, as hereinabove described, whereby such cells express lung surfactant protein.
  • the vector is transfected into the cells, followed by infection of the cells with the encapsidation- defective helper virus.
  • the adenoviral construction shuttle plasmid pAvS6 was constructed in several steps using standard cloning techniques including polymerase chain reaction based cloning techniques.
  • Ad-dl327 (Thimmappaya, et al., Cell, Vol. 31, pg.
  • adenovirus 543 (1983) is identical to adenovirus 5 except that an Xbal fragment including base ⁇ 28593 to 30470 (or map units 78.5 to 84.7) of the adenovirus 5 genome, and which is located in the E3 region, has been deleted.
  • Genbank accession #M73260 The comlete Adenovirus 5 genome is registered as Genbank accession #M73260, incorporated herein by reference, and the virus is available from the American Type Culture Collection, Rockville, Maryland, U.S.A. under accession number VR-5.
  • Ad-dl327 was constructed by routine methods from Adenovirus 5 (Ad5). The method is outlined briefly as follows and previously described by Jones and Shenk, Cell 13:181-188 (1978).
  • Ad5 DNA is isolated by proteolytic digestion of the virion and partially cleaved with Xba I restriction endonuclease. The Xba I fragments are then reassembled by ligation as a mixture of fragments. This results in some ligated genomes with a sequence similar to Ad5, except excluding sequences 28593 bp to 30470 bp.
  • This DNA is then transfected into suitable cells (e.g. KB cells, HeLa cells, 293 cells) and overlaid with soft agar to allow plaque formation.
  • the ITR, encapsidation signal, Rous Sarcoma Virus promoter, the adenoviral tripartite leader (TPL) sequence and linking sequences were assembled as a block using PCR amplification ( Figure 2).
  • the ITR and encapsidation signal (sequences 1-392 of Ad-dl327 [identical to sequences from Ad5, Genbank accession #M73260]) were amplified (amplification 1) together from Ad-dl327 using primers containing NotI or Ascl restriction sites.
  • the Rous Sarcoma Virus LTR promoter was amplified (amplification 2) from the plasmid pRC/RSV (sequences 209 to 605; Invitrogen, San Diego, CA) using primers containing an Ascl site and an Sfil site. DNA products from amplifications 1 and 2 were joined using the "overlap" PCR method (amplification 3) with only the NotI primer and the Sfil primer. Complementarity between the Ascl containing end of each initial DNA amplification product from reactions 1 and 2 allowed joining of these two pieces during amplification.
  • the TPL was amplified (amplification 4) (sequences 6049 to 9730 of Ad-dl327 [identical to similar sequences from Ad5, Genbank accession #M73260]) from cDNA made from mRNA isolated from 293 cells infected for 16 hours with Ad-dl327 using primers containing Sfil and Xbal sites respectively. DNA fragments from amplification reactions 3 and 4 were then joined using PCR (amplification 5) with the NotI- and Xbal-site-containing primers, thus creating . the complete gene block.
  • amplification 4 sequences 6049 to 9730 of Ad-dl327 [identical to similar sequences from Ad5, Genbank accession #M73260]
  • DNA fragments from amplification reactions 3 and 4 were then joined using PCR (amplification 5) with the NotI- and Xbal-site-containing primers, thus creating . the complete gene block.
  • the ITR-encapsidation signal-TPL fragment was then purified, cleaved with NotI and Xbal and inserted into the NotI, Xbal cleaved pHR plasmid.
  • This plasmid was designated pAvS6A and the orientation was such that the NotI site of the fragment was next to the T7 RNA polymerase site ( Figure 3).
  • the SV40 early polyA signal was removed from SV40 DNA as an Hpal-BamHI fragment, treated with T4 DNA polymerase and inserted into the Sail site of the plasmid pAvS6A-( Figure 3) to create pAvS6 ( Figures 3 and 4).
  • pAVS6SPB#7 Such clones are named pAVS6SPB#7, pAVS6SPB#12, and pAVS6SPB#13.
  • pAVS6SPB#7 is shown in Figure 7. The orientation of the SPB DNA within the shuttle plasmid was obtained by evaluating the DNA sequences of the two termini of the SPB cDNA insert in the plasmid with primers derived from pAVS6.
  • the recombinant adenoviral vector AV1SPB1 ( Figure 8), containing SPB cDNA was constructed through homologous recombination between the Ad5 deletion mutant Ad-dl327 ( Figure 8), and pAVS6SPB#7. Homologous recombination, or "crossing over,” occurs between Ad-dl327 and pAVS6.SPB#7, along the segment common to both Ad-dl327 and pAVS6.SPB#7 which corresponds to bases 3328 to 6241 (or map units 9.24 to 17.34) of the adenovirus 5 genome.
  • Ad-dl327 has a deleted E3 region in which base pairs 28593 to 30470 are absent (Thimmappaya, et al. Cell, Vol. 31, pgs. 543-551 (1982)).
  • pAVS6SPB#7 contains an adenoviral 5' ITR, an origin of replication contained completely within the 5' ITR, an Ela enhancer and encapsidation signal, a Rous Sarcoma Virus promoter, an adenovirus 5 tripartite leader sequence and the 2 b human SPB cDNA including the entire protein coding sequence (nucleotide ⁇ 1 to 1172), and the SV40 poly A signal.
  • Example 2 293 cells (ATCC No. CRL 1573) were infected with AV1SPB1 at a multiplicity of infection (MOI) of 50 MOI units. At 12 hours post-infection, the cells were radiolabeled with 3S S-methionine (50 ⁇ Ci/ml) overnight. Identical amounts of labeled protein were used for immunoprecipitation with antisera against SPB. Immunopreciptates were analyzed by SDS-polyacrylamide gel electrophoresis on 16% gel. The gels were fluorographed. C 14 - labeled molecular weight markers and BioRad broad range molecular weight markers were used as size markers.
  • MOI multiplicity of infection
  • lane 1 shows an uninfected control
  • lane 2 show ⁇ AVISPBl-infected 293 cells in which electrophoresis of immunoprecipitates occurred under reducing conditions
  • lane 3 shows AVISPBl-infected 293 cells in which electrophoresis of immunoprecipitates occurred under non-reducing conditions.
  • the precursor protein also was detected in both reduced and unreduced conditions.
  • Example 3 Mouse lung Type II-like epithelial cell lines were transduced with adenoviral vector AVISPBI, which contains the full length human SPBcDNA under the control of the Rous Sarcoma Virus promoter. Expression of SPB was assessed by RNa ⁇ e protection using 32 P-labelled probes specific for endogenous mouse SPB (upper band) or human SPB (middle band). A B-tubulin specific probe (lowest band) also was used to ensure that the same amount of RNA was added to each assay. The B-tubulin probe did not recognize human B-tubulin in lane 1.
  • the filter was crosslinked (UV Crosslink, Stratagene), and hybridized with a 32 P-labeled 2.0 kb human SP-B cDNA probe prepared by random priming (Loftstrand) and evaluated by autoradiography. Lungs from uninfected control rats and from rats infected with AvlLacZ4 also were subjected to the above hybridization procedure.
  • AvlLacZ4 treated rats and untreated rats were used as controls.
  • de novo synthesis and secretion of human SP-B peptide was detected from lung fragments removed from animals infected with AvlSPBl, and was not detected in uninfected animals or in animals infected with AvlLacZ4.
  • Human SP-B peptide was detected as secreted, with 8 kda and 18 kda oligo eric forms suggesting that vector derived precursor SP-B (proSP-B) was processed after in vivo adenoviral vector-mediated gene transfer.
  • the animals were sacrificed 48 hours after infection, and the lungs were prepared for in situ hybridization analysis according to the method of Wert, et al.. Development Biology. Vol. 156, pgs. 426-443 (1993), using human SP-BcRNA. An uninfected rat was used as a control.
  • ADDRESSEE Carella, Byrne, Bain, Gilfillan,
  • CTACTCCGTC ATCCTGCTCG ACACGCTGCT GGGCCGCATG CTGCCCCAGC TGGTCTGCCG 840
  • GAGCTATTGC TTTGTTAAGA TATAAAAAGG GGTTTCTTTT TGTCTTTCTG TAAGGTGGAC 1860

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pulmonology (AREA)
  • Physics & Mathematics (AREA)
  • Virology (AREA)
  • Toxicology (AREA)
  • Plant Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention se rapporte à un vecteur adénoviral renfermant une séquence d'ADN codant une protéine surfactant des poumons. Le vecteur adénoviral peut être un vecteur adénoviral déficient de réplication qui est exempt au moins de la majorité des séquences d'ADN E1 et E3. Ces vecteurs peuvent être utilisés dans la génération de particules virales infectieuses capables de transduire des cellules épithéliales des poumons in vivo afin de faciliter l'expression de la protéine surfactant des poumons par ces cellules.
PCT/US1994/003831 1993-04-08 1994-04-07 Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons WO1994023582A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP6523310A JPH09500782A (ja) 1993-04-08 1994-04-07 肺界面活性タンパク質をコード化するｄｎａを含むアデノウイルスベクター
EP94914075A EP0701401A4 (fr) 1993-04-08 1994-04-07 Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US4440693A 1993-04-08 1993-04-08
US08/044,406 1993-04-08

Publications (2)

Publication Number Publication Date
WO1994023582A1 true WO1994023582A1 (fr) 1994-10-27
WO1994023582A9 WO1994023582A9 (fr) 1994-12-08

Family

ID=21932217

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1994/003831 WO1994023582A1 (fr) 1993-04-08 1994-04-07 Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons

Country Status (4)

Country Link
EP (1) EP0701401A4 (fr)
JP (1) JPH09500782A (fr)
CA (1) CA2160136A1 (fr)
WO (1) WO1994023582A1 (fr)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0710123A1 (fr) * 1993-06-07 1996-05-08 The Regents of The University of Michigan Vecteurs d'adenovirus pour therapie genique
WO1996020732A2 (fr) * 1994-12-30 1996-07-11 Chiron Corporation Administration atraumatique de vehicules d'apport de genes
WO1998032860A1 (fr) * 1997-01-28 1998-07-30 Baxter International Inc. Procede de production a haut rendement de vecteurs adenoviraux
WO1999002647A2 (fr) * 1997-07-10 1999-01-21 Hepavec Ag Für Gentherapie Vecteurs de clonage pour preparer des adenovirus de type virus minimaux
WO1999027101A1 (fr) * 1997-11-25 1999-06-03 Princeton University Procede de preparation de vecteurs d'adenovirus, vecteurs ainsi prepares et leurs utilisations
EP0927263A1 (fr) * 1996-01-05 1999-07-07 Genetic Therapy, Inc. Generation de vecteurs adenoviraux a mediation de recombinase
US5994128A (en) * 1995-06-15 1999-11-30 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6265212B1 (en) 1995-06-15 2001-07-24 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
EP1123383A1 (fr) * 1998-10-20 2001-08-16 Children's Hospital Medical Center La proteine tensioactive d servant au diagnostic et au traitement de l'emphyseme pulmonaire
US6403370B1 (en) 1997-02-10 2002-06-11 Genstar Therapeutics Corporation Oncolytic/immunogenic complementary-adenoviral vector system
US6670188B1 (en) 1998-04-24 2003-12-30 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6783980B2 (en) 1995-06-15 2004-08-31 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6838428B2 (en) 1998-10-20 2005-01-04 Children's Hospital Medical Center Surfactant protein D for the prevention and diagnosis of pulmonary emphysema
US9370555B2 (en) 1998-10-20 2016-06-21 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury
US9492503B2 (en) 2011-02-04 2016-11-15 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2705686B1 (fr) * 1993-05-28 1995-08-18 Transgene Sa Nouveaux adénovirus défectifs et lignées de complémentation correspondantes.

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5585362A (en) * 1989-08-22 1996-12-17 The Regents Of The University Of Michigan Adenovirus vectors for gene therapy
CA2592997A1 (fr) * 1992-12-03 1994-06-09 Genzyme Corporation Vecteurs pseudo-adenoviraux
FR2705686B1 (fr) * 1993-05-28 1995-08-18 Transgene Sa Nouveaux adénovirus défectifs et lignées de complémentation correspondantes.
FR2707664B1 (fr) * 1993-07-13 1995-09-29 Centre Nat Rech Scient Vecteurs viraux et utilisation en thérapie génique.

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
Cell, Volume 68, issued 10 January 1992, M.A. ROSENFELD et al., "In Vivo Transfer of the Human Cystic Fibrosis Transmembrane Conductance Regulator Gene to the Airway Epithelium", pages 143-155, see the entire document. *
DNA, Volume 8, No. 2, issued 1989, T.J. PILOT-MATIAS et al., "Structure and Organization of the Gene Encoding Human Pulmonary Surfactant Proteolipid SP-B", pages 75-86, see the entire document. *
Journal of Clinical Investigation, Volume 81, issued March 1988, S.D. REVAK et al., "Use of Human Surfactant Low Molecular Weight Apoproteins in the Reconstitution of Surfactant Biologic Activity", pages 826-833, see the entire document. *
Science, Volume 252, issued 19 April 1991, M.A. ROSENFELD et al., "Adenovirus-Mediated Transfer of a Recombinant Alpha1-Antitrypsin Gene to the Lung Epithelium in Vivo", pages 431-434, see the entire document. *
See also references of EP0701401A4 *
W. DOERFLER, "Adenovirus DNA", published 1986, by Martinus Nijhoff Publishing (Boston), pages 53-95, see the entire document. *

Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0710123A1 (fr) * 1993-06-07 1996-05-08 The Regents of The University of Michigan Vecteurs d'adenovirus pour therapie genique
EP0710123A4 (fr) * 1993-06-07 1998-07-29 Univ Michigan Vecteurs d'adenovirus pour therapie genique
WO1996020732A2 (fr) * 1994-12-30 1996-07-11 Chiron Corporation Administration atraumatique de vehicules d'apport de genes
WO1996020732A3 (fr) * 1994-12-30 1996-12-12 Viagene Inc Administration atraumatique de vehicules d'apport de genes
US6602706B1 (en) 1995-06-15 2003-08-05 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6692966B2 (en) 1995-06-15 2004-02-17 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6238893B1 (en) 1995-06-15 2001-05-29 Introgene B.V. Method for intracellular DNA amplification
US6395519B1 (en) 1995-06-15 2002-05-28 Introgene B.V. Means and methods for nucleic acid delivery vehicle design and nucleic acid transfer
US6783980B2 (en) 1995-06-15 2004-08-31 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6306652B1 (en) 1995-06-15 2001-10-23 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US5994128A (en) * 1995-06-15 1999-11-30 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6033908A (en) * 1995-06-15 2000-03-07 Introgene, B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6265212B1 (en) 1995-06-15 2001-07-24 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
EP0927263A1 (fr) * 1996-01-05 1999-07-07 Genetic Therapy, Inc. Generation de vecteurs adenoviraux a mediation de recombinase
EP0927263A4 (fr) * 1996-01-05 1999-07-07
US6156497A (en) * 1996-01-05 2000-12-05 Genetic Therapy, Inc. Recombinase-mediated generation of adenoviral vectors
WO1998032860A1 (fr) * 1997-01-28 1998-07-30 Baxter International Inc. Procede de production a haut rendement de vecteurs adenoviraux
US6403370B1 (en) 1997-02-10 2002-06-11 Genstar Therapeutics Corporation Oncolytic/immunogenic complementary-adenoviral vector system
WO1999002647A2 (fr) * 1997-07-10 1999-01-21 Hepavec Ag Für Gentherapie Vecteurs de clonage pour preparer des adenovirus de type virus minimaux
WO1999002647A3 (fr) * 1997-07-10 1999-04-15 Hepavec Ag Fuer Gentherapie Vecteurs de clonage pour preparer des adenovirus de type virus minimaux
WO1999027101A1 (fr) * 1997-11-25 1999-06-03 Princeton University Procede de preparation de vecteurs d'adenovirus, vecteurs ainsi prepares et leurs utilisations
US6878549B1 (en) 1998-04-24 2005-04-12 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6670188B1 (en) 1998-04-24 2003-12-30 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US7037716B2 (en) 1998-04-24 2006-05-02 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
EP1123383A1 (fr) * 1998-10-20 2001-08-16 Children's Hospital Medical Center La proteine tensioactive d servant au diagnostic et au traitement de l'emphyseme pulmonaire
US6838428B2 (en) 1998-10-20 2005-01-04 Children's Hospital Medical Center Surfactant protein D for the prevention and diagnosis of pulmonary emphysema
EP1123383A4 (fr) * 1998-10-20 2002-07-17 Childrens Hosp Medical Center La proteine tensioactive d servant au diagnostic et au traitement de l'emphyseme pulmonaire
EP1995315A3 (fr) * 1998-10-20 2010-06-23 Children's Hospital Medical Center Protéine D tensioactive pour la prévention et le diagnostic d'emphysème pulmonaire
US9370555B2 (en) 1998-10-20 2016-06-21 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury
US9492503B2 (en) 2011-02-04 2016-11-15 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury
US10159716B2 (en) 2011-02-04 2018-12-25 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury
US10632178B2 (en) 2011-02-04 2020-04-28 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury

Also Published As

Publication number Publication date
CA2160136A1 (fr) 1994-10-27
EP0701401A1 (fr) 1996-03-20
JPH09500782A (ja) 1997-01-28
EP0701401A4 (fr) 1997-07-16

Similar Documents

Publication Publication Date Title
US5935935A (en) Adenoviral vectors for treatment of hemophilia
JP3565859B2 (ja) 改良されたアデノウイルスおよびその使用法
EP0696206B1 (fr) Adenovirus contenant des proteines fibreuses modifiees
Schiedner et al. Genomic DNA transfer with a high-capacity adenovirus vector results in improved in vivo gene expression and decreased toxicity
US5824544A (en) Adenovirus vectors for gene therapy
RU2219241C2 (ru) Дефектный рекомбинантный аденовирусный вектор (варианты)
CA2278616C (fr) Adenovirus comprenant des proteines d'hexon modifiees
WO1994023582A1 (fr) Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons
JP3755827B2 (ja) 組み込み可能な組み換えアデノウィルス、それらの製造及びそれらの治療的利用
AU727992B2 (en) Adenoviral vectors comprising a modified E4 region but retaining E4ORF3
WO1994023582A9 (fr) Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons
JP4733795B2 (ja) ヒツジアデノウイルスベクターを用いた遺伝子治療
JPH10510987A (ja) 改良アデノウイルスベクターおよび生産者細胞
Yei et al. Adenoviral-mediated gene transfer of human surfactant protein B to respiratory epithelial cells.
AU718772B2 (en) Adenovirus vectors for gene therapy
WO1996014837A1 (fr) Therapie genique contre l'hypercholesterolemie
US20020019051A1 (en) Chimeric adenoviral vectors
JP2007530004A (ja) 疾患を処置するためのサブグループｂアデノウイルスベクター
CN112041437A (zh) 用于恢复f8基因功能的腺相关病毒组合物和其使用方法
WO1996036365A1 (fr) Therapie genique de carcinomes hepatocellulaires par expression de genes specifiques du cancer
Connelly Adenoviral vectors

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

COP Corrected version of pamphlet

Free format text: PAGES 1/23,DESCRIPTION,REPLACED BY NEW PAGES 1-19;PAGES 24-26,CLAIMS,REPLACED BY NEW PAGES 20-22;PAGES 1/12-12/12,DRAWINGS,REPLACED BY NEW PAGES BEARING THE SAME NUMBER

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 2160136

Country of ref document: CA

Ref document number: 1994914075

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 1996 532557

Country of ref document: US

Date of ref document: 19960118

Kind code of ref document: A

WWP Wipo information: published in national office

Ref document number: 1994914075

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1994914075

Country of ref document: EP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载