Transgenic animals expressing heparanase and uses thereof

• US 20030217375A1
• Filed: 02/24/2003
• Published: 11/20/2003
• Est. Priority Date: 08/31/1998
• Status: Abandoned Application

• Current Assignee: Hadasit Medical Research Services & Development Ltd (Hadassah, The Women's Zionist Organization of America, Inc.), Insight Strategy & Marketing Ltd.
• Original Assignee: Hadasit Medical Research Services & Development Ltd (Hadassah, The Women's Zionist Organization of America, Inc.)
• Inventors: Metzger, Shula, Chajek-Shaul, Tova, Pecker, Iris, Vlodavsky, Israel, Ilan, Neta, Zcharia, Eyal, Goldshmidt, Orit
• Application Number:
  US10/371,218
• Publication Number:
  US 20030217375A1
• Time in Patent Office: Days
• US Class Current: 800/7
• 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: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/01166 Heparanase (3.2.1.166)

Abstract

A transgenic non-human animal expressing heparanase from a transgene, methods for its preparation, compositions-of-matter derived therefrom and uses thereof.

Claims

1. A transgenic non-human animal whose genome comprises an exogenous polynucleotide sequence integrated into said genome, said exogenous polynucleotide sequence including a promoter active in tissues of the non-human, and a region encoding a human heparanase, wherein said promoter and said region encoding human heparanase are operably linked in said exogenous polynucleotide such that human heparanase is expressed in at least a portion of the cells of the non-human animal.

2. The transgenic non-human animal of claim 1, being homozygous for said exogenous polynucleotide sequence.

3. The transgenic non-human animal of claim 1, being heterozygous for said exogenous polynucleotide sequence.

4. The transgenic non-human animal of claim 1, having a single locus harboring said exogenous polynucleotide sequence.

5. The transgenic non-human animal of claim 1, having at least two loci each harboring said exogenous polynucleotide sequence.

6. The transgenic non-human animal of claim 1, wherein said human heparanase is genetically modified to be cleavable into an active form via a protease.

7. The transgenic non-human animal of claim 1, wherein said heparanase is processed by an endogenous protease of the non-human animal into an active form.

8. The transgenic non-human animal of claim 1, wherein said region of said exogenous polynucleotide sequence encodes an active form of heparanase.

9. The transgenic non-human animal of claim 1, being a mammal.

10. The transgenic non-human animal of claim 1, being an avian.

11. The transgenic non-human animal of claim 1, wherein said exogenous polynucleotide sequence includes a tissue specific promoter for directing expression of said heparanase in a tissue specific manner.

12. The transgenic non-human animal of claim 1, wherein said promoter is a constitutive promoter for directing expression of said heparanase in constitutive manner.

13. The transgenic non-human animal of claim 1, wherein said promoter is an inducible promoter for directing expression of said heparanase in an inducible manner.

14. The transgenic non-human animal of claim 1, wherein said promoter is selected from the group consisting of beta-lactoglobulin promoter, Rb promoter, preproendothelin-1 promoter, beta-actin promoter, TetO promoter, metallothionein promoter, whey acidic protein (WAP) promoter, casein promoter and lactalbumin promoter.

15. The transgenic non-human mammal of claim 9, wherein said heparanase is expressed in, and secreted by, cells of mammary glands of said mammal.

16. The transgenic avian of claim 10, wherein said promoter is selected from the group consisting of chicken lyzozyme promoter, cytomegalovirus promoter and chicken immunoglobulin promoter.

17. The transgenic non-human avian of claim 10, wherein said heparanase is expressed in, and secreted by, egg producing cells of said avian.

18. Sex cells derived from the transgenic non-human animal of claim 1.

19. Semen derived from the transgenic non-human animal of claim 1.

20. An embryo derived from the transgenic non-human animal of claim 1.

21. A composition of matter comprising milk derived from a non-human transgenic mammal, said milk having detectable human heparanase activity.

22. A composition of matter comprising egg yolk and/or white from a transgenic avian, said egg yolk and/or white having detectable human heparanase activity.

23. A method of producing recombinant human heparanase, the method comprising the steps of:

(a) obtaining a transgenic non-human mammal having mammary glands, whose genome comprises an exogenous polynucleotide sequence integrated into said genome, said exogenous polynucleotide sequence including a promoter active in tissues of the non-human mammal, and a region encoding a human heparanase, wherein said promoter and said region encoding human heparanase are operably linked in said exogenous polynucleotide such that the recombinant human heparanase is secreted into milk being produced by said mammary glands;

(b) milking said non-human mammal so as to obtain milk containing the recombinant human heparanase; and

(c) purifying the recombinant human heparanase from said milk.

24. The method of claim 23, wherein said promoter active in tissues of said non-human mammal is a milk protein gene promoter.

25. The method of claim 24, wherein said milk protein gene promoter is selected from the group consisting of beta-lactoglobulin promoter, Rb promoter, preproendothelin-1 promoter, whey acidic protein (WAP) promoter, casein promoter and lactalbumin promoter.

26. A method of producing recombinant human heparanase, the method comprising the steps of:

(a) obtaining a transgenic female avian having egg producing cells whose genome comprises an exogenous polynucleotide sequence integrated into said genome, said exogenous polynucleotide sequence including a promoter active in tissues of said transgenic female avian, and a region encoding a human heparanase, wherein said promoter and said region encoding human heparanase are operably linked in said exogenous polynucleotide such that the recombinant human heparanase is secreted into eggs being produced by said egg producing cells;

(b) collecting eggs laid by said transgenic female avian so as to obtain eggs containing the recombinant human heparanase; and

(c) purifying the recombinant human heparanase from said eggs.

27. The method of claim 26, wherein said promoter active in tissues of said transgenic female avian is an egg protein gene promoter.

28. The method of claim 27, wherein said egg protein gene promoter is selected from the group consisting of chicken lyzozyme promoter and chicken immunoglobulin promoter.