Modulating cancer cell characteristics

• US 7,666,676 B2
• Filed: 05/16/2006
• Issued: 02/23/2010
• Est. Priority Date: 05/16/2005
• Status: Expired due to Fees

• Current Assignee: St. Joseph's Children's Hospital
• Original Assignee: St. Joseph's Children's Hospital
• Inventors: Braccili, Selena, Lawman, Michael J. P.
• Primary Examiner(s): ANGELL, JON E
• Application Number:
  US11/436,962
• Publication Number:
  US 20070161588A1
• Time in Patent Office: 1,379 Days
• Field of Search: 435/325, 435/440, 536/24.5
• US Class Current: 435/440
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: C07K 14/46 from vertebrates

Abstract

The hedgehog pathway in cerebellar cancer cells was modulated with siRNA specifically targeted to the shh and gli-I genes. Silencing of the two genes in a medullablastoma cell line transfected with the siRNAs caused significant reduction of mRNA specific for the targeted shh and gli-I genes and a loss of protein expression. The disclosed methods and compositions may be useful for treatment of a range of primitive neuroectodermal tumors (PNET) by shutting down or modulating the expression of gene products associated with the hedgehog pathway.

Claims

1. A method for inhibiting expression of SHh in primitive neuroectodermal (PNET) tumor cells, comprising transfecting the tumor cells with an effective amount of a siRNA primer pair consisting of the antisense shh gene sequences SEQ ID NO:

6 and SEQ ID NO;

8 wherein the transfected tumor cells change from non-adherent proliferating cells to adherent proliferating cells after transfecting in vitro thereby altering growth, phenotype and growth characteristics of the adherent proliferating cells.

2. The method of claim 1 wherein the PNET tumor cell is selected from the group consisting of medulloblastoma, basal cell carcinoma, fibroma, meningioma, and rhabdomyosarcoma.

3. The method of claim 1 wherein the PNET tumor cell is a D283 cell.

4. The method of claim 1 wherein inhibiting expression of the shh gene decreases-protein expression by up to about 85% compared to normal protein expression in the tumor cells.

5. The method of claim 1 wherein the transfected tumor cells exhibit reduced growth in vitro compared to untransfected tumor cells.