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DE102006051623A1 - Gene transfer into target cells in vivo comprises applying a gene transfer vector and a transduction enhancer at the same time and place - Google Patents

Gene transfer into target cells in vivo comprises applying a gene transfer vector and a transduction enhancer at the same time and place Download PDF

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DE102006051623A1
DE102006051623A1 DE102006051623A DE102006051623A DE102006051623A1 DE 102006051623 A1 DE102006051623 A1 DE 102006051623A1 DE 102006051623 A DE102006051623 A DE 102006051623A DE 102006051623 A DE102006051623 A DE 102006051623A DE 102006051623 A1 DE102006051623 A1 DE 102006051623A1
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Peter A Horn
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    • 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
    • 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
    • A61K48/0083—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the administration regime
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/17011—Spumavirus, e.g. chimpanzee foamy virus
    • C12N2740/17041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/17043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

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Abstract

Gene transfer into target cells in vivo comprises applying a gene transfer vector and a transduction enhancer at the same time and place. Independent claims are also included for: (1) nonhuman mammal genetically modified by the above method; (2) the mixture of materials used in the above method. ACTIVITY : Anti-AIDS; Antidiabetic; Antianemic; Cytostatic. MECHANISM OF ACTION : Gene therapy.

Description

Stand der TechnikState of the art

Hämatopoetische Stammzellen sind besonders attraktive Zielzellen für gentherapeutische Ansätze. In den letzten drei Jahren konnte die Stammzellgentherapie endlich den Beweis erbringen, dass mit diesem Ansatz genetische Erkrankungen wie der X-linked SCID (ein schwerer Immundefekt) klinisch geheilt werden können. Im Rahmen dieser Studie kam es jedoch in zwei Fällen zu schwerwiegenden Nebenwirkungen, der Entwicklung einer leukämie-ähnlichen Erkrankung, mutmaßlich durch eine Insertionsmutagenese. Dadurch ist deutlich geworden, dass unser Wissen zu den Mechanismen und Nebenwirkungen noch sehr lückenhaft ist und dass technische Verbesserungen im Bereich der Gentransfermethodik nötig sind. Der ex vivo Gentransfer in hämatopoetische Stammzellen beinhaltet typischerweise die Gewinnung von Stammzellen aus dem Knochenmark durch Knochenmarkpunktion oder (nach Wachstumfaktorgabe) aus dem peripheren Blut mittels Apherese. Die Progenitoren werden meistens mittels CD34 Selektion angereichert und in Gegenwart von verschiedenen Wachstumsfaktoren kultiviert, um eine Aktivierung oder Proliferation der Zielzellen zu erreichen. Für retrovirale Vektoren ist eine Mitose für die Vektorintegration unerlässlich. Die dafür notwendige, relativ lange ex vivo Kultur kann jedoch zu einem verminderten Stammzellengraftment (Anwachsen) führen [Sellers 1999]. Nach ex vivo Transduktion der Zielzellen werden diese dem Empfänger intravenös appliziert. Um ein Engraftment der so behandelten Stammzellen zu erreichen, ist eine myeloablative Behandlung des Empfängers nötig, da die infundierten Stammzellen mit den verbleibenden endogenen Stammzellen konkurrieren. Ansätze zum ex vivo Gentransfer haben demnach eine Reihe von potentiellen Limitationen und Nachteilen:

  • – Die notwendige myeloablative Behandlung des autologen Empfängers ist sehr nebenwirkungsreich und demnach in vielen klinischen Situationen, insbesondere für nicht-maligne Erkrankungen nicht vertretbar.
  • – Bei manchen Erkrankungen (z. B: Fanconi-Anämie), die gentherapeutisch behandelt werden könnten, sind die Stammzellen so empfindlich, dass sie ex vivo praktisch nicht manipulierbar und demnach nicht transduzierbar sind.
  • – Eine Steigerung der Zahl genveränderter Zellen durch multiple Applikation ist aufgrund des relativ aufwendigen und belastenden Verfahrens nicht möglich.
  • – Die Entfernung der hämatopoetischen Stammzellen aus ihrer natürlichen Umgebung verändert mutmaßlich ihre einzigartigen biologischen Eigenschaften in Bezug auf Proliferation und Selbsterneuerung.
  • – Die ex vivo Exposition von hämatopoetischen Stammzellen mit rekombinanten Wachstumsfaktoren vermindert ihre Fähigkeiten zu einem Homing ins Knochenmark und Engraftment.
  • – Die derzeitig verfügbaren Protokolle für ex vivo Gentransfer sind relativ ineffizient.
Hematopoietic stem cells are particularly attractive target cells for gene therapy approaches. In the last three years, stem cell gene therapy has finally proven that this approach can clinically cure genetic disorders such as X-linked SCID (a severe immunodeficiency). However, in two studies, severe side effects such as the development of leukemia-like disease, presumably due to insertion mutagenesis, occurred in this study. This has made it clear that our knowledge of the mechanisms and side effects is still very patchy and that technical improvements in the field of gene transfer methodology are needed. The ex vivo gene transfer into hematopoietic stem cells typically involves the recovery of stem cells from the bone marrow by bone marrow puncture or (after growth factor delivery) from the peripheral blood via apheresis. The progenitors are mostly enriched by CD34 selection and cultured in the presence of various growth factors to achieve activation or proliferation of the target cells. For retroviral vectors, mitosis is essential for vector integration. However, the relatively long ex vivo culture required for this can lead to decreased stem cell grafting (growth) [Sellers 1999]. After ex vivo transduction of the target cells they are administered to the recipient intravenously. In order to engraft the stem cells so treated, myeloablative treatment of the recipient is necessary as the infused stem cells compete with the remaining endogenous stem cells. Approaches to ex vivo gene transfer therefore have a number of potential limitations and disadvantages:
  • - The necessary myeloablative treatment of the autologous recipient is very rich in side effects and therefore unacceptable in many clinical situations, especially for non-malignant diseases.
  • - For some diseases (eg Fanconi anemia) that could be treated with gene therapy, the stem cells are so sensitive that they can not be manipulated ex vivo and are therefore not transduceable.
  • - An increase in the number of gene-modified cells by multiple application is not possible due to the relatively complex and stressful process.
  • The removal of hematopoietic stem cells from their natural environment presumably alters their unique biological properties in terms of proliferation and self-renewal.
  • Ex vivo exposure of hematopoietic stem cells to recombinant growth factors reduces their ability to homing to bone marrow and engraftment.
  • The currently available protocols for ex vivo gene transfer are relatively inefficient.

Aufgrund dieser inherenten Limitationen von ex vivo Gentransfer haben verschiedene Arbeitsgruppen bereits Ansätze zur in vivo Transduktion hämatopoetischer Stammzellen durchgeführt. Nelson et al. [1997] applizierten bestrahlte retrovirale Verpackungszellinien ohne weitere Konditionierung in den Femur (Oberschenkelknochen) von Kaninchen. Genveränderte Blutzellen konnten bis zu 20 Monate nach Injektion nachgewiesen werden. Im Rahmen einer ähnlichen Studie von Porada et al. [2000] wurde unkonzentrierter retroviraler Vektor in das Darmbein neugeborener Schafe gespritzt. Obgleich das injizierte Volumen des Vektors in den meisten Tieren bei nur 2 ml (Spanne: 1–6) lag, konnte das Transgen bis zu über ein Jahr lang nachgewiesen werden.by virtue of These inherent limitations of ex vivo gene transfer have different Working groups already have approaches to in vivo transduction hematopoietic stem cells. Nelson et al. [1997] applied irradiated retroviral packaging cell lines without further conditioning in the femur (thighbone) of rabbits. Gene-modified blood cells could reach up to Be detected 20 months after injection. As part of a similar Study by Porada et al. [2000] became unfocused retroviral Vector injected into the ilium of newborn sheep. Although that injected volumes of the vector in most animals at only 2 ml (Span: 1-6), the transgene could be up to about proven for a year.

Um die Effizienz und die Sicherheit von verschiedenen Ansätzen der Stammzellgentherapie beurteilen zu können, sind Studien in klinisch relevanten Tiermodellen, insbesondere in Primaten, der akzeptierte Goldstandard. Auch im Primatenmodell gibt es bereits Daten zum in vivo Gentransfer in hämatopoietische Stammzellen. Newbound et al. [2001] injizierten retrovirale Verpackungszellinien in das Knochenmark von insgesamt sechs Rhesusaffen (Macaca mulatta) ohne weitere Konditionierung. Die initial relativ hohe Frequenz transduzierter Zellen verringerte sich rapide und eine humorale Immunantwort gegen Komponenten des viralen Vektors wurde nachgewiesen. Trotzdem stieg die Frequenz transduzierter Zellen nach einer zweiten Injektion von Verpackungszellinien erneut deutlich an, was verdeutlichte, dass eine multiple Applikation prinzipiell durchführbar ist.Around the efficiency and safety of different approaches to be able to assess stem cell gene therapy are studies in clinically relevant animal models, especially in primates, the accepted gold standard. There are already in the primate model Data for in vivo gene transfer into hematopoietic stem cells. Newbound et al. [2001] injected retroviral packaging cell lines into the bone marrow of a total of six rhesus monkeys (Macaca mulatta) without further conditioning. The initially relatively high frequency transduced cells decreased rapidly and a humoral Immune response against components of the viral vector was detected. Nevertheless, the frequency of transduced cells increased after a second Injection of packaging cell lines, clearly demonstrating that a multiple application is feasible in principle.

Alle aufgeführten Veröffentlichungen applizierten jedoch virale Vektoren ohne gleichzeitig Transduktionsfrequenz verbessernde Substanzen wie beispielsweise hämatopoetische Wachstumsfaktoren gleichzeitig oder zeitnah zu applizieren.All However, publications listed applied viral vectors without simultaneously improving transduction frequency Substances such as hematopoietic growth factors apply simultaneously or in a timely manner.

Zusammenfassend kann also festgehalten werden, dass es bereits eine Reihe von wissenschaftlichen Publikationen und von Patenten gibt, die sich mit in vivo Gentransfer befassen, jedoch ist in allen derzeitig publizierten Arbeiten zu diesem Thema die Effizienz eines solchen in vivo Gentransfers sehr niedrig.In summary So it can be stated that there are already a number of scientific Publications and patents exist dealing with in vivo gene transfer However, all currently published works are closed In this topic, the efficiency of such in vivo gene transfer is very low.

Aufgabe der ErfindungObject of the invention

Das zu lösende Problem liegt wie oben dargestellt im wesentlichen in der niedrigen Gentransfereffizienz bei einem in vivo Gentransfer sowie der Immunantwort gegen den benutzten Vektor bzw. das Transgen.The problem to be solved is as shown above essentially in the low gene transfer efficiency in an in vivo gene transfer and the Immune response against the used vector or the transgene.

Eine signifikante Verbesserung der Transduktionsrate viraler und anderer Vektoren und eine signifikante Verminderung der Immunogenität der applizierten Gentransfervektoren kann durch gleichzeitige oder zeitnahe lokale Applikation von bestimmten chemisch definierten Substanzen erreicht werden. Hierzu gehören insbesondere

  • – (rekombinant hergestellte) Wachstumsfaktoren, die die Zellen zu Wachstum anregen oder sie sonstig stimulieren
  • – Substanzen, die eine Verbesserung der Virusbindung an die Zielzellen ermöglichen (z. B. Fibronektin, Protaminsulfat, CH-296)
  • – Substanzen, die eine Verlängerung der Aufenthaltszeit der Vektoren am Ort der Zielzellen ermöglichen (wie Protaminsulfat)
  • – Substanzen, die eine Verminderung der Immunogenität des eingesetzten Gentransfervektors oder eine verminderte Immunantwort bewirken (wie Corticoide, Immunsuppresiva, siRNA gegen bestimmte Nukleotidsequenzen zwecks Expressionsverminderung, z. B. von antigenpräsentierenden Molekülen wie HLA).
A significant improvement in the transduction rate of viral and other vectors and a significant reduction in the immunogenicity of the applied gene transfer vectors can be achieved by simultaneous or timely local application of certain chemically defined substances. These include in particular
  • - (recombinantly produced) growth factors that stimulate or otherwise stimulate the cells to grow
  • - Substances that enhance virus binding to target cells (eg fibronectin, protamine sulfate, CH-296)
  • Substances which allow prolongation of the residence time of the vectors at the target cell site (such as protamine sulfate)
  • Substances which cause a reduction in the immunogenicity of the gene transfer vector used or a reduced immune response (such as corticoids, immunosuppresives, siRNA against specific nucleotide sequences for the purpose of reducing expression, for example of antigen-presenting molecules such as HLA).

Es geht bei der hier vorgestellten Erfindung demnach nicht um eine gänzlich neue Substanz, sondern den Einsatz bekannter chemischer/pharmazeutischer Substanzen zur signifikanten Verbesserung einer im Prinzip bereits bekannten Technik (in vivo Gentransfer).It Accordingly, the present invention is not concerned with one completely new substance, but the use of known chemical / pharmaceutical Substances to significantly improve one in principle already known technique (in vivo gene transfer).

Literaturverzeichnisbibliography

  • 1. Link CJ Jr, Seregina T, Levy JP, Martin M, Ackermann M, Moorman DW. Murine retroviral vector producer cells survival and toxicity in the dog liver. In Vivo. 2000 Sep–Oct; 14(5): 643–9 1. Link CJ Jr, Seregina T, Levy JP, Martin M, Ackermann M, Moorman DW. Murine retroviral vector producer cells survival and toxicity in the dog liver. In vivo. 2000 Sep-Oct; 14 (5): 643-9
  • 2. Nelson DM, Metzger ME, Donahue RE, Morgan RA. In vivo retrovirus-mediated gene transfer into multiple hematopoietic lineages in rabbits without preconditioning. Hum Gene Ther. 1997 Apr 10; 8(6): 747–54 Second Nelson DM, Butcher ME, Donahue RE, Morgan RA. In vivo retrovirus-mediated gene transfer into multiple hematopoietic lineages in rabbits without preconditioning. Hum Gene Ther. 1997 Apr 10; 8 (6): 747-54
  • 3. Newbound GC, Cooper JR, O'Rourke JP, Baskin CR, Bunnell BA. Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells. Exp Hematol. 2001 Feb; 29(2): 163–73 Third Newbound GC, Cooper JR, O'Rourke JP, Baskin CR, Bunnell BA. Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells. Exp Hematol. 2001 Feb; 29 (2): 163-73
  • 4. Porada CD, Tran ND, Zhao Y, Anderson WF, Zanjani ED. Neonatal gene therapy. transfer and expression of exogenous genes in neonatal sheep following direct injection of retroviral vectors into the bone marrow space. Exp Hematol. 2000 Jun; 28(6): 642–50 4th Porada CD, Tran ND, Zhao Y, Anderson WF, Zanjani ED. Neonatal gene therapy. The expression of exogenous genes in neonatal sheep is followed by direct injection of retroviral vectors into the bone marrow space. Exp Hematol. 2000 Jun; 28 (6): 642-50
  • Patentschriften zum Thema in vivo Gentransfer:Patents on in vivo gene transfer:
  • 5. CA000002115262A [EN] In-Vivo Gene Transfer Using Implanted Retroviral Producer Cells5th CA000002115262A [EN] In vivo Gene Transfer Using Implanted Retroviral Producer Cells
  • 6. EP000000602118A4 [EN]-i(IN VIVO) GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS. ...6th EP000000602118A4 [EN] -i (IN VIVO) GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS. ...
  • 7. EP000000602118A1 [EN]-i(IN VIVO) GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS. ...7th EP000000602118A1 [EN] -i (IN VIVO) GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS. ...
  • 8. US000006663858B1 [] In Vivo gene transfer using implanted retroviral producer cells8th. US000006663858B1 [In vivo gene transfer using implanted retroviral producer cells
  • 9. US000006663858B1 [] In vivo gene transfer using implanted retroviral producer cells,9th US000006663858B1 In vivo gene transfer using implanted retroviral producer cells,
  • 10. WO001993004167A1 [EN] IN VIVO GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS10th WO001993004167A1 [EN] IN VIVO GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS
  • 11. WO9738729 IN VIVO GENE TRANSFER METHODS FOR WOUND HEALING11th WO9738729 IN VIVO GENE TRANSFER METHODS FOR WOUND HEALING
  • 12. EP0938904 Mammalian cell transduction for use in gene therapy12th EP0938904 Mammalian cell transduction for use in gene therapy
  • 13. DE0069232497T2 BEHANDLUNG VON KRANKHEITEN DURCH ORTSSPEZIFISCHE INSTILLATION VON ZELLEN ODER ORTSSPEZIFISCHE TRANSFORMATION VON ZELLEN SOWIE AUSRUESTUNG DAFUER13th DE0069232497T2 TREATMENT OF ILLNESSES BY ORIGINAL INSTALLATION OF CELLS OR ORGANIC TRANSFORMATION OF CELLS AND EQUIPMENT THEREOF
  • 14. WO002003062383A2 [EN] IN VIVO GENE TRANSFER14th WO002003062383A2 [EN] IN VIVO GENE TRANSFER
  • 15. WO002003062383A3 [EN] IN VIVO GENE TRANSFER15th WO002003062383A3 [EN] IN VIVO GENE TRANSFER
  • 16. WO002003066104A2 [EN] PHARMACEUTICAL COMPOSITION WHICH IMPROVES IN VIVO GENE TRANSFER16th WO002003066104A2 [EN] PHARMACEUTICAL COMPOSITION WHICH IMPROVES IN VIVO GENE TRANSFER

ZITATE ENTHALTEN IN DER BESCHREIBUNGQUOTES INCLUDE IN THE DESCRIPTION

Diese Liste der vom Anmelder aufgeführten Dokumente wurde automatisiert erzeugt und ist ausschließlich zur besseren Information des Lesers aufgenommen. Die Liste ist nicht Bestandteil der deutschen Patent- bzw. Gebrauchsmusteranmeldung. Das DPMA übernimmt keinerlei Haftung für etwaige Fehler oder Auslassungen.This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Zitierte PatentliteraturCited patent literature

  • - CA 000002115262 A [0008] - CA 000002115262 A [0008]
  • - EP 000000602118 A4 [0008] EP 000000602118 A4 [0008]
  • - EP 000000602118 A1 [0008] EP 000000602118 A1 [0008]
  • - US 000006663858 B1 [0008, 0008] US 000006663858 B1 [0008, 0008]
  • - WO 001993004167 A1 [0008] WO 001993004167 A1 [0008]
  • - WO 9738729 [0008] - WO 9738729 [0008]
  • - EP 0938904 [0008] EP 0938904 [0008]
  • - DE 0069232497 T2 [0008] - DE 0069232497 T2 [0008]
  • - WO 002003062383 A2 [0008] WO 002003062383 A2 [0008]
  • - WO 002003062383 A3 [0008] WO 002003062383 A3 [0008]
  • - WO 002003066104 A2 [0008] WO 002003066104 A2 [0008]

Zitierte Nicht-PatentliteraturCited non-patent literature

  • - Link CJ Jr, Seregina T, Levy JP, Martin M, Ackermann M, Moorman DW. Murine retroviral vector producer cells survival and toxicity in the dog liver. In Vivo. 2000 Sep–Oct; 14(5): 643–9 [0008] - Link CJ Jr, Seregina T, Levy JP, Martin M, Ackermann M, Moorman DW. Murine retroviral vector producer cells survival and toxicity in the dog liver. In vivo. 2000 Sep-Oct; 14 (5): 643-9 [0008]
  • - Nelson DM, Metzger ME, Donahue RE, Morgan RA. In vivo retrovirus-mediated gene transfer into multiple hematopoietic lineages in rabbits without preconditioning. Hum Gene Ther. 1997 Apr 10; 8(6): 747–54 [0008] - Nelson DM, Butcher ME, Donahue RE, Morgan RA. In vivo retrovirus-mediated gene transfer into multiple hematopoietic lineages in rabbits without preconditioning. Hum Gene Ther. 1997 Apr 10; 8 (6): 747-54 [0008]
  • - Newbound GC, Cooper JR, O'Rourke JP, Baskin CR, Bunnell BA. Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells. Exp Hematol. 2001 Feb; 29(2): 163–73 [0008] - Newbound GC, Cooper JR, O'Rourke JP, Baskin CR, Bunnell BA. Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells. Exp Hematol. 2001 Feb; 29 (2): 163-73 [0008]
  • - Porada CD, Tran ND, Zhao Y, Anderson WF, Zanjani ED. Neonatal gene therapy. transfer and expression of exogenous genes in neonatal sheep following direct injection of retroviral vectors into the bone marrow space. Exp Hematol. 2000 Jun; 28(6): 642–50 [0008] - Porada CD, Tran ND, Zhao Y, Anderson WF, Zanjani ED. Neonatal gene therapy. The expression of exogenous genes in neonatal sheep is followed by direct injection of retroviral vectors into the bone marrow space. Exp Hematol. 2000 Jun; 28 (6): 642-50 [0008]

Claims (20)

In vivo Gentransfer in Zielzellen dadurch gekennzeichnet, dass in engem zeitlichen Zusammenhang die Transduktionfrequenz erhöhende Substanzen am gleichen Ort wie die Gentransfervektoren appliziert werden.In vivo gene transfer into target cells, characterized in that the transduction frequency-increasing substances in the same place as the gene transfer vectors are applied in close temporal relation. Verfahren nach Anspruch 1, wobei es sich bei den Zielzellen um hämatopoetische Stammzellen handelt.The method of claim 1, wherein the Target cells are hematopoietic stem cells. Verfahren nach Anspruch 1, wobei es sich bei den Zielzellen um Stromazellen handelt.The method of claim 1, wherein the Target cells are stromal cells. Verfahren nach Anspruch 3, wobei es sich bei den Zielzellen um Stromazellen des Knochenmarks handelt.The method of claim 3, wherein the Target cells are stromal cells of the bone marrow. Verfahren nach Anspruch 1, wobei die Transduktionfrequenz erhöhende Substanzen rekombinante Wachstumsfaktoren umfasst.The method of claim 1, wherein the transduction frequency increasing substances comprises recombinant growth factors. Verfahren nach Anspruch 1, wobei die Transduktionfrequenz erhöhende Substanzen Protaminsulfat umfasst.The method of claim 1, wherein the transduction frequency increasing substances comprises protamine sulfate. Verfahren nach Anspruch 1, wobei die Transduktionfrequenz erhöhende Substanzen Fibronektin umfasst.The method of claim 1, wherein the transduction frequency increasing substances comprises fibronectin. Verfahren nach Anspruch 1, wobei die Transduktionfrequenz erhöhende Substanzen das Fibronektinfragment CH-296 umfasst.The method of claim 1, wherein the transduction frequency increasing substances comprises the fibronectin fragment CH-296. Verfahren nach Anspruch 5, wobei die rekombinanten Wachstumsfaktoren SCF umfasst.The method of claim 5, wherein the recombinant Growth factors SCF includes. Verfahren nach Anspruch 5, wobei die rekombinanten Wachstumsfaktoren G-CSF umfasst.The method of claim 5, wherein the recombinant Growth factors G-CSF includes. Verfahren nach Anspruch 5, wobei die rekombinanten Wachstumsfaktoren Flt3-L umfasst.The method of claim 5, wherein the recombinant Growth factors Flt3-L includes. Verfahren nach Anspruch 5, wobei die rekombinanten Wachstumsfaktoren TPO umfasst.The method of claim 5, wherein the recombinant Growth factors TPO includes. Verfahren nach Anspruch 1, wobei es sich bei den Gentransfervektoren um retrovirale Vektoren handelt.The method of claim 1, wherein the Gene transfer vectors are retroviral vectors. Verfahren nach Anspruch 1, wobei es sich bei den Gentransfervektoren um lentivirale Vektoren handelt.The method of claim 1, wherein the Gene transfer vectors for lentiviral vectors. Verfahren nach Anspruch 1, wobei es sich bei den Gentransfervektoren um Foamy-Virus-Vektoren handelt.The method of claim 1, wherein the Gene transfer vectors to foamy virus vectors. Verfahren nach Anspruch 1, wobei es sich bei den Gentransfervektoren um adenovirale Vektoren handelt.The method of claim 1, wherein the Gene transfer vectors are adenoviral vectors. Verfahren nach Anspruch 1, wobei es sich bei den Gentransfervektoren um AAV-Vektoren handelt.The method of claim 1, wherein the Gene transfer vectors to AAV vectors. Verwendung eines Verfahrens gemäß Anspruch 1–17 zur Herstellung eines Arzneimittels zur individuellen Gentherapie bei einem Immundefektsyndrom, einer Stoffwechselerkrankung, bei Blutbildungsstörung oder bei einer malignen oder infektiösen Erkrankung.Use of a method according to claim 1-17 for the manufacture of a medicament for the individual Gene therapy for an immune deficiency syndrome, a metabolic disease, in case of blood formation disorder or in a malignant or infectious Illness. Ein Säugetier außer dem Menschen, welches durch ein Verfahren gemäß Anspruch 1–17 gentechnisch verändert wurde.A mammal other than humans, which by a method according to claim 1-17 was genetically modified. Das Stoffgemisch, dass in einem Verfahren gemäß Anspruch 1–17 eingesetzt wird.The mixture of substances that in a process according to claim 1-17 is used.
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993004167A1 (en) 1991-08-13 1993-03-04 The Regents Of The University Of California In vivo gene transfer using implanted retroviral producer cells
WO1997038729A1 (en) 1996-04-12 1997-10-23 The Regents Of The University Of Michigan In vivo gene transfer methods for wound healing
EP0938904A1 (en) 1998-02-09 1999-09-01 Leuven Research & Development vzw Mammalian cell transduction for use in gene therapy
DE69232497T2 (en) 1991-06-28 2002-11-14 The Regents Of The University Of Michigan, Ann Arbor TREATMENT OF DISEASES BY LOCAL INSTILLATION OF CELLS OR LOCALLY SPECIFIC TRANSFORMATION OF CELLS AND EQUIPMENT THEREFOR
WO2003062383A2 (en) 2002-01-16 2003-07-31 Baylor College Of Medicine In vivo gene transfer
WO2003066104A2 (en) 2002-02-08 2003-08-14 Institut National De La Sante Et De La Recherche Medicale (Inserm) Pharmaceutical composition which improves in vivo gene transfer

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE69232497T2 (en) 1991-06-28 2002-11-14 The Regents Of The University Of Michigan, Ann Arbor TREATMENT OF DISEASES BY LOCAL INSTILLATION OF CELLS OR LOCALLY SPECIFIC TRANSFORMATION OF CELLS AND EQUIPMENT THEREFOR
WO1993004167A1 (en) 1991-08-13 1993-03-04 The Regents Of The University Of California In vivo gene transfer using implanted retroviral producer cells
CA2115262A1 (en) 1991-08-13 1993-03-04 David Barba In-vivo gene transfer using implanted retroviral producer cells
EP0602118A1 (en) 1991-08-13 1994-06-22 The Regents Of The University Of California $i(IN VIVO) GENE TRANSFER USING IMPLANTED RETROVIRAL PRODUCER CELLS
EP0602118A4 (en) 1991-08-13 1996-02-28 Univ California GENE -i TRANSFER (IN VIVO) USING IMPLANTED CELLS PRODUCING A RETROVIRUS.
US6663858B1 (en) 1991-08-13 2003-12-16 The Regents Of The University Of California In Vivo gene transfer using implanted retroviral producer cells
WO1997038729A1 (en) 1996-04-12 1997-10-23 The Regents Of The University Of Michigan In vivo gene transfer methods for wound healing
EP0938904A1 (en) 1998-02-09 1999-09-01 Leuven Research & Development vzw Mammalian cell transduction for use in gene therapy
WO2003062383A2 (en) 2002-01-16 2003-07-31 Baylor College Of Medicine In vivo gene transfer
WO2003066104A2 (en) 2002-02-08 2003-08-14 Institut National De La Sante Et De La Recherche Medicale (Inserm) Pharmaceutical composition which improves in vivo gene transfer

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Link CJ Jr, Seregina T, Levy JP, Martin M, Ackermann M, Moorman DW. Murine retroviral vector producer cells survival and toxicity in the dog liver. In Vivo. 2000 Sep-Oct; 14(5): 643-9
Nelson DM, Metzger ME, Donahue RE, Morgan RA. In vivo retrovirus-mediated gene transfer into multiple hematopoietic lineages in rabbits without preconditioning. Hum Gene Ther. 1997 Apr 10; 8(6): 747-54
Newbound GC, Cooper JR, O'Rourke JP, Baskin CR, Bunnell BA. Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells. Exp Hematol. 2001 Feb; 29(2): 163-73
Porada CD, Tran ND, Zhao Y, Anderson WF, Zanjani ED. Neonatal gene therapy. transfer and expression of exogenous genes in neonatal sheep following direct injection of retroviral vectors into the bone marrow space. Exp Hematol. 2000 Jun; 28(6): 642-50

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