Protein modulators of resistance to alkylating agents

• US 7,875,274 B2
• Filed: 12/13/2006
• Issued: 01/25/2011
• Est. Priority Date: 12/16/2005
• Status: Expired due to Fees

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Bredel, Markus, Sikic, Branimir
• Primary Examiner(s): Long; Scott
• Application Number:
  US11/638,161
• Publication Number:
  US 20070148145A1
• Time in Patent Office: 1,504 Days
• Field of Search: None
• US Class Current: 424/93.21
• 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 2600/118 Prognosis of disease development
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: Y10S 977/802 Virus-based particle

Abstract

There is disclosed a method for identifying a therapeutically responsive phenotype, as distinguished, e.g. from an alkylating agent resistant phenotype in a cell, which method may be used to evaluate the likelihood of successful outcome of treating a tumor cell with an alkylating agent. The method is directed to the NF-κB activation in response to DNA damage caused by alkylating agents. It comprises the step of measuring a level of expression of a protein, which participates in the NF-κB pathway. Preferably it comprises measuring the expression of TNFAIP3 in the cell, wherein a resistant phenotype has less expression of TNFAIP3 than a sensitive phenotype. Another particularly significant gene, which predicts survival, is NFKBIA. Other genes whose altered expression level is associated with resistance or prognosis are TNIP1, TNIP2, RIP, NFKBIB, Beta4GalNAc-T4, NFKBIE, C8orf4, LIF, CD44, FBXO32, and SDC1, and these are also measured in certain embodiments.

Claims

1. A method for identifying a therapeutically responsive phenotype in a brain tumor cell, comprising the step of measuring a level of expression of TNFAIP3 gene, NFKBIA gene, TNIP1 gene and TNIP2 genes in the cell and identifying said cell as being sensitive or resistant to an alkylating agent based on said level of expression of TNFAIP3 gene, NFKBIA gene, TNIP1 gene, and TNIP2 genes, wherein a responsive phenotype has more expression of TNIP1, TNFAIP3, NFKBIA and TNIP2 than a resistant phenotype.

2. The method of claim 1 wherein the level of expression of the gene is measured by determining a level of mRNA encoding the gene.

3. The method of claim 2 wherein the measurement of mRNA comprises the step of real-time reverse transcription PCR.

4. The method of claim 2 wherein the measurement of mRNA comprises the step of using a microarray.

5. The method of claim 1 where the measuring of the expression of the gene comprises determining a level of the cognate protein in the cell.

6. The method of claim 1 further comprising the step of measuring the level of expression of at least three genes selected from the group consisting of C8orf4, MGMT, Beta4GalNAc-T4, SDC1, CD44, FBXO32, RIP, NFKBIE, and NFKBIB.

7. A method for identify an alkylating agent resistance phenotype in a brain tumor cell, comprising the step of measuring a level of expression of a TNFAIP3 gene in the cell, wherein a resistant phenotype is identified us having said alkylating agent resistance phenotype based on less expression of TNFAIP3 than a sensitive phenotype and said measuring includes measurement of expression of a control gene, which is selected from the group consisting of:

PBGD (porphobilinogen deaminase). 5-aminolevulinate synthase (ALAS), beta actin, and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

8. A method for assessing probability of survival time of a patient with a brain tumor, comprising:

obtaining a cell from the tumor; and

measuring, a level of expression of genes TNFAIP3, SDC1, CD44 and FBXO32 in the cell, wherein shortened survival is based on a lowered level of expression of all four genes.

9. The method of claim 8 further comprising the step of measuring the level of expression of at least one gene selected from the group consisting of NFKBIA, C8orf4, LIF, MGMT, Beta4GalNAc-T4, RIP, TNIP1, NFKBIE, NFKBIB, and TNIP2.

10. The method of claim 9 wherein the measurement of level of expression comprises the measurement of mRNA levels using a microarray.