SHIP-Deficiency to Increase Megakaryocyte Progenitor Production

• US 20100260730A1
• Filed: 06/21/2010
• Published: 10/14/2010
• Est. Priority Date: 11/20/2003
• Status: Active Grant

• Current Assignee: University of South Florida (State University System of Florida)
• Original Assignee: University of South Florida (State University System of Florida)
• Inventors: Desponts, Caroline, Perez, Lia Elena, KERR, WILLIAM G.
• Granted Patent: US 8,008,273 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2217/206 Animal model comprising tissue-specific expression system, e.g. tissue specific expression of transgene, of Cre recombinase
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/0387 Animal model for diseases of the immune system
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)

Abstract

The invention concerns a method for increasing megakaryocyte and megakaryocyte progenitor numbers in vitro or in vivo by suppressing SH2-containing inositol-5-phosphatase (SHIP) function in megakaryocytes or megakaryocyte progenitors expressing the SHIP gene. SHIP function can be suppressed by administering an interfering RNA, or other SHIP inhibitor, to the megakaryocytes or megakaryocyte progenitors in vitro or in vivo.

Claims

1. A method for improving haematopoietic recovery in a patient in need thereof, comprising:

harvesting megakaryocytes or megakaryocyte progenitors from a patient;

contacting the megakaryocytes or megakaryocyte progenitors with an effective amount of an inhibitor of SH2-containing inositol-5-phosphatase (SHIP) function, thereby expanding the megakaryocytes or megakaryocyte progenitors; and

reinfusing the expanded megakaryocytes or megakaryocyte progenitors into the patient.

2. The method of claim 1, wherein said method further comprises determining the amount or concentration of megakaryocytes or megakaryocyte progenitors in the patient before said harvesting, after said reinfusing, or both.

3. The method of claim 2, wherein the inhibitor of SHIP function is selected from the group consisting of interfering RNA, antisense oligonucleotide, ribozyme, DNAzyme, nucleic acid modifier, PNA, nonstandard nucleic acid, aptamer, decoy, and dominant/negative mutant.

4. The method of claim 3, wherein the inhibitor of SHIP function is an interfering RNA targeted to SHIP mRNA.

5. The method of claim 4, wherein the interfering RNA is targeted to the SHIP enzymatic domain (inositol 5′

-phosphatase domain).

6. The method of claim 4, wherein the interfering RNA is targeted to the amino-terminal src-homology domain (SH2).

7. The method of claim 1, wherein the patient is human.

8. The method of claim 4, wherein the interfering RNA targets a region within the SHIP mRNA encoded by a nucleotide sequence selected from the group consisting of GCCTGTTGTCATCCATTGA (SEQ ID NO:

3), ATAAGTTGGTGATCTTGGT (SEQ ID NO;

4), GCCACATCTGTACTGACAA (SEQ ID NO;

5), AGACAGGCATTGCAAACAC (SEQ ID NO;

6), ACATCACTCACCGCTTCAC (SEQ ID NO;

7), TCTTAACTACCGTGTGGAT (SEQ ID NO;

8), AATACGCCTACACCAAGCA (SEQ ID NO;

9), GTACCAGCGACATCATGAC (SEQ ID NO;

10), GCGACATCATGACGAGTGA (SEQ ID NO;

11), AGGACAGATTGAGTTTCTC (SEQ ID NO;

12), GGTGCTATGCCACATTGAA (SEQ ID NO;

13), GTTTGGTGAGACTCTTCCA (SEQ ID NO;

14), AGACGGAGCGTGATGAATC (SEQ ID NO;

15), GCTTCCAGAAGAGCATCTTAT (SEQ ID NO;

20), and GCCCATATCACCCAAGAAGTTT (SEQ ID NO;

21).

9. The method of claim 4, wherein the interfering RNA comprises a nucleotide sequence selected from the group consisting of SEQ TD NO:

16, SEQ ID NO;

17, SEQ ID NO;

18, and SEQ ID NO;

19.