MICRO RNAS AS MARKERS OF THE FUNCTIONAL STATE OF A DENDRITIC CELL

• US 20100266555A1
• Filed: 11/14/2008
• Published: 10/21/2010
• Est. Priority Date: 11/14/2007
• Status: Abandoned Application

• Current Assignee: Bioneer Corporation, DanDrit Biotech A/S (Renovaro Biosciences, Inc.)
• Original Assignee: Bioneer Corporation, DanDrit Biotech A/S (Renovaro Biosciences, Inc.)
• Inventors: Zocca, Mai-Britt, Wakatsuki Pedersen, Ayako, Skjøde Jensen, Simon, Holmstrøm, Kim
• Application Number:
  US12/741,801
• Publication Number:
  US 20100266555A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/178 miRNA, siRNA or ncRNA

Abstract

The present invention relates to the use of specified micro RNAs as markers of the functional state of a dendritic cell. In one aspect the invention relates to a method for producing a quality-controlled therapeutic composition comprising dendritic cells. In another aspect the invention relates to a method of in vitro screening of immunomodulatory compounds.

Claims

1. A method for monitoring the functional state of dendritic cells, comprising:

comparing an expression profile of one or more micro RNAs (miRNAs) which comprises a sequence that has at least 70% identity to a nucleic acid sequence represented by one of SEQ ID NOs;

1 to 45 to an expression profile of a reference sequence from a standard population of dendritic cells;

analyzing differential regulation of the one or more miRNAs to determine the functional state of the dendritic cells.

2. The method according to claim 1, further comprising distinguishing between immature and immunogenic dendritic cells, wherein the sequence is represented by one of SEQ ID NOs:

1 to 36 and the micro RNA is used as markers to distinguish between immature and immunogenic dendritic cells.

3. The method according to claim 2, wherein the sequence is represented by one of SEQ ID NOs:

1, 4, 5, 7, 9, 18, 26, 27, 28, 29, 30 and 31.

4. The method according to claim 3, wherein the sequence is represented by one of SEQ ID NOs:

4, 7, 26 and 27.

5. The method according to claim 1, further comprising distinguishing between immature and tolergenic dendritic cells, wherein the sequence is represented by one of SEQ ID NOs:

26 to 45 and the micro RNA is used as markers to distinguish between immature and tolerogenic dendritic cells.

6. The method according to claim 5, wherein the sequence is represented by done of SEQ ID NOs:

26, 27, 32, 34, 37, 39, 41 and 42.

7. The method according to claim 6, wherein the sequence is represented by one of SEQ ID NOs:

27 and 41.

8. The method according to claim 1, further comprising distinguishing between immature dendritic cells and tolerogenic or immunogenic dendritic cells, wherein the sequence is represented by one of SEQ ID NOs:

26 to 36 and the micro RNA is used as markers to distinguish between immature dendritic cells and tolerogenic or immunogenic dendritic cells.

9. The method according to claim 1, wherein the sequence has at least 90% identity to the reference nucleic acid sequence.

10. The method according to claim 1, wherein at least two different micro RNAs are used.

11. The method according to claim 1, wherein at least three different micro RNAs are used.

12. The method according to claim 1, where the dendritic cell is of human origin.

13. The method according to claim 1, wherein the dendritic cells being monitored are in therapeutic compositions comprising dendritic cells.

14. The method according to claim 13, wherein the therapeutic composition is a vaccine.

15. The method according to claim 13, wherein the therapeutic composition is used therapeutically for adoptive transfer.

16. A method of producing a quality controlled composition comprising dendritic cells, by which the functional state of the dendritic cells in the composition is monitored, the method comprising:

a) extracting nucleic acids from the dendritic cells from the composition,

b) monitoring the expression profile of one or more micro RNAs comprising a sequence that has at least 70% identity to a nucleic acid sequence selected from the group consisting of SEQ IDs 1 to 45 extracted in step a), and

c) comparing the expression profile obtained in b) with a standard profile from immature dendritic cells.

17. The method according to claim 16, wherein the sequence is represented by one of SEQ ID NOs:

1 to 36.

18. The method according to claim 17, wherein the sequence is represented by to one of SEQ ID NOs:

1, 4, 5, 7, 9, 18, 26, 27, 28, 29, 30 and 31.

19. The method according to claim 18, wherein the sequence is represented by to one of SEQ ID NOs:

4, 7, 26 and 27.

20. The method according to claim 16, wherein the sequence is represented by one of SEQ ID NOs:

26 to 45.

21. The method according to claim 20, wherein the sequence is represented by one of SEQ ID NOs:

26, 27, 32, 34, 37, 39, 41 and 42.

22. The method according to claim 21, wherein the sequence is represented by one of SEQ ID NOs:

27 and 41.

23. The method according to claim 16, wherein the sequence is represented by one of SEQ ID NOs:

26 to 36.

24. A quality-controlled therapeutic composition obtainable by the method of claims 16.

25-28. -28. (canceled)

29. A method for in vitro screening for compounds having immunomodulatory effect, comprising:

a) providing a test population of immature dendritic cells;

b) producing, from the test population, a micro RNA expression profile of at least one micro RNA comprising a sequence that is essentially homologous to a sequence represented by one of SEQ ID NOs;

1 to 45;

c) contacting the population of dendritic cells with a test compound;

d) producing a micro RNA expression profile as in step b); and

e) comparing the expression profiles obtained in step b) and step d), where a significant difference in profile will be indicative of immunomodulatory effect of the test compound.

30. The method according to claim 29, wherein the micro RNAs comprise a sequence that is essentially homologous to a sequence represented by one of SEQ ID NOs:

1 to 36.

31. The method according to claim 29, wherein the micro RNAs comprise a sequence that is essentially homologous to a sequence represented by one of SEQ ID NOs:

26 to 45.

32. The method according to claim 29, wherein the micro RNAs comprise a sequence that is essentially homologous to a sequence represented by one of SEQ ID NOs:

26 to 36.