Methods for genetic modification of hematopoietic progenitor cells and uses of the modified cells

• US 7,345,025 B2
• Filed: 07/10/2002
• Issued: 03/18/2008
• Est. Priority Date: 07/10/2001
• Status: Expired due to Fees

• Current Assignee: Johnson & Johnson Research Pty Ltd. (Johnson & Johnson)
• Original Assignee: Johnson & Johnson Research Pty Ltd. (Johnson & Johnson)
• Inventors: Gerlach, Wayne, Symonds, Geoffrey P., Sun, Lun-Quan, MacPherson, Janet L., Amado, Rafael G., Fanning, Gregory C.
• Primary Examiner(s): ZARA, JANE J
• Application Number:
  US10/192,980
• Publication Number:
  US 20040072771A1
• Time in Patent Office: 2,078 Days
• Field of Search: 514/44, 424/93.1, 435/320.1, 435/6, 435/91.1, 435/91.31, 435/455, 435/458, 435/375, 536/23.1, 536/24.5
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: C12N 15/1132 against retroviridae, e.g. HIV
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/121 Hammerhead
• CPC Class Codes: C12N 2310/14 interfering N.A.

Abstract

Described are compositions and methods relating to gene therapy, particularly as applied to hematopoietic progenitor (HP) cells, to transduced cells and methods of obtaining them, and to methods of using them to provide prolonged engraftment of modified hematopoietic cells in human subjects. The invention particularly relates to ex vivo gene therapy of HP cells for treatment or prevention of HIV infection.

Claims

1. A composition for engraftment of transduced autologous CD34⁺ hematopoietic progenitor cells in a human subject which comprises a pharmaceutically acceptable carrier and a minimum dose of 0.52×

10⁶ transduced CD34⁺ hematopoietic progenitor cells per kg body weight of the subject in a total CD34⁺ cell population of at least 1.63×

10⁶ CD34⁺ hematopoietic progenitor cells per kg body weight of the subject, the transduced cells comprising a RRz2 vector encoding a ribozyme and the number of transduced cells being determinable by performing competitive PCR on such transduced CD34⁺ hematopoietic cells, wherein the ribozyme is encoded by DNA nucleotides having the sequence 5′

-TTA GGA TCC TGA TGA GTC CGT GAG GAC GAA ACT GGC TC-3′

(SEQ ID NO;

     3).

2. The composition of claim 1, comprising a minimum dose of 5×

10⁶ transduced CD34⁺ hematopoietic progenitor cells per kg body weight of the subject in a total CD34⁺ cell population comprising 9.37×

10⁶ CD34⁺ hematopoietic progenitor cells per kg body weight of the subject.

3. The composition of claim 1, wherein the CD34⁺ hematopoietic progenitor cells are from mobilized peripheral blood of the subject.

4. The composition of claim 1, wherein the cells are transduced in the presence of an agent that enhances co-localization of the cells with a viral construct.

5. The composition of claim 4, wherein the agent is a recombinant fragment of fibronectin.

6. The composition of claim 5, wherein the recombinant fragment of fibronectin is the CH296 fragment of human fibronectin.

7. The composition of claim 2, wherein the CD34⁺ hematopoietic progenitor cells are from mobilized peripheral blood of the subject.

8. The composition of claim 2, wherein the cells are transduced in the presence of a CH296 fragment of human fibronectin.

9. The composition of claim 3, wherein the cells are transduced in the presence of a CH296 fragment of human fibronectin.

10. The composition of claim 7, wherein the cells are transduced in the presence of a CH296 fragment of human fibronectin.

11. The composition of claim 1, wherein the composition is substantially free of cytokines.

12. The composition of claim 1, wherein the composition is substantially free of virus.

13. The composition of claim 1, wherein in the subject the transduced CD34⁺ hematopoietic progenitor cells give rise to progeny cells for at least 12 months.

14. A composition for engraftment of transduced autologous CD34⁺ hematopoietic progenitor cells in a human subject which comprises a pharmaceutically acceptable carrier and a minimum dose of 0.5×

10⁶ transduced CD34⁺ hematopoietic progenitor cells per kg body weight of the subject in a total CD34⁺ cell population of at least 1.7×

10⁶ CD34⁺ hematopoietic progenitor cells per kg body weight of the subject, the transduced cells comprising a RRz2 vector encoding a ribozyme and the number of transduced cells being determinable by performing competitive PCR on such transduced CD34⁺ hematopoietic cells, wherein the ribozyme is encoded by DNA nucleotides having the sequence 5′

-TTA GGA TCC TGA TGA GTC CGT GAG GAC GAA ACT GGC TC-3′

(SEQ ID NO;

     3).

15. The composition of claim 14, comprising a minimum dose of 5×

10⁶ transduced CD34⁺ hematopoietic progenitor cells per kg body weight of the subject.

16. The composition of claim 14, wherein the CD34⁺ hematopoietic progenitor cells are from mobilized peripheral blood of the subject.

17. The composition of claim 14, wherein the cells are transduced in the presence of an agent that enhances co-localization of the cells with a retroviral construct.

18. The composition of claim 17, wherein the agent is a CH296 fragment of human fibronectin.