GENES ASSOCIATED WITH CHEMOTHERAPY RESPONSE AND USES THEREOF

• US 20100284915A1
• Filed: 06/28/2007
• Published: 11/11/2010
• Est. Priority Date: 06/30/2006
• Status: Abandoned Application

• Current Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Original Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Inventors: Zhang, Chunsheng, Loboda, Andrey, Dai, Hongyue
• Application Number:
  US12/307,114
• Publication Number:
  US 20100284915A1
• Time in Patent Office: Days
• US Class Current: 424/9.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/118 Prognosis of disease development
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/57415 of breast
• CPC Class Codes: G01N 33/57449 of ovaries
• CPC Class Codes: G01N 33/57484 involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• CPC Class Codes: G16B 20/00 ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
• CPC Class Codes: G16B 20/20 Allele or variant detection, e.g. single nucleotide polymorphism [SNP] detection
• CPC Class Codes: G16B 25/00 ICT specially adapted for hybridisation; ICT specially adapted for gene or protein expression
• CPC Class Codes: G16B 25/10 Gene or protein expression profiling; Expression-ratio estimation or normalisation
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

The invention provides molecular markers that are associated with responsiveness of a cancer patient to a chemotherapy treatment, and methods and computer systems for determining such responsiveness based on measurements of these molecular markers. The present invention also provides methods and compositions for enhancing the efficacy of chemotherapies in patients by modulating the expression or activity of genes encoding these molecular markers and/or their encoded proteins.

Claims

1. A method for predicting the responsiveness of a mammalian patient having a cancer to a chemotherapy regimen, comprising:

predicting said mammalian patient (a) as responsive to said chemotherapy regimen, if expression and/or activity of one or more gene products in a cell sample taken from said mammalian patient is not up-regulated relative to a reference population of individuals of the same species as said mammalian patient;

or (b) as non-responsive to said chemotherapy regimen, if expression and/or activity of said one or more gene products is up-regulated relative to said reference population of individuals, wherein said one or more gene products comprise respectively products of one or more different genes selected from the group consisting of genes corresponding to SEQ ID NOs;

1-39 or respective functional equivalents thereof.

2. The method of claim 1, further comprising:

determining, prior to said predicting step, whether expression and/or activity of said one or more gene products is up-regulated as relative to said reference population of individuals.

3. The method of claim 2, wherein said determining step is carried out by a method comprising:

determining one or more chemotherapy response scores (CR scores) based on measurements of said one or more gene products in said cell sample, wherein said one or more CR scores indicate whether expression and/or activity of said one or more gene products is up-regulated as compared to individuals in said reference population.

4. The method of claim 3, comprising:

determining a CR score that is an average of said measurements of said one or more gene products, wherein said mammalian patient is predicted as responsive if said average is equal to a predetermined threshold value or as non-responsive if said average is greater than said predetermined threshold value.

5. The method of claim 3, comprising:

determining a first CR score that is a first measurement of a gene product of a gene having the greatest expressive range among a first subset of said one or more different genes, wherein said first subset is selected from the group consisting of genes having SEQ ID NOs;

1-19 or

determining a second CR score that is a second measurement of a gene product of a gene having the greates expressive range among a second subset of said one or more different genes, wherein said second subset is selected from the group consisting of genes having SEQ ID NOs;

20-39, wherein said mammalian patient is predicted as responsive if said first or second measurement is less or equal to a predetermined threshold value or as non-responsive if said first or second measurement is greater than said predetermined threshold value.

6. The method of claim 3, wherein said step of determining one or more CR scores is carried out by a method comprising:

(a1) comparing a marker profile comprising said measurements of said one or more gene products with a responsive template and/or a non-responsive template, wherein said responsive template comprises measurements of said one or more gene products representative of measurements of said one or more genes products in a plurality of mammalian patients being responsive to said chemotherapy regimen, and said non-responsive template comprises measurements of said one or more gene products representative of measurements of said plurality of genes products in a plurality of mammalian patients being non-responsive to said chemotherapy regimen; and

(a2) determining a first degree of similarity between said marker profile and said responsive template and/or a second degree of similarity between said marker profile and said non-responsive template, wherein said first and second degrees of similarity are said one or more CR scores, and wherein said mammalian patient is (b1) predicted to be responsive if said first degree of similarity is greater than said second degree of similarity or if said first degree of similarity is greater than a predetermined threshold or (b2) predicted to be non-responsive if said first degree of similarity is no greater than said second degree of similarity or if said second degree of similarity is no greater than said predetermined threshold.

7. The method of claim 6, wherein said first or second degree of similarity is represented by a correlation coefficient between said marker profile and said respective template.

8. The method of claim 6, wherein the measurement of each gene product in said responsive template is an average of the measurements of said gene product in a plurality of responsive mammalian patients, and wherein the measurement of each gene product in said non-responsive template is an average of the measurements of said gene product in a plurality of non-responsive mammalian patients.

9. The method of claim 3, wherein said step of determining one or more CR scores is carried out by a method comprising using a chemotherapy response classifier selected from the group consisting of an artificial neural network (ANN) classifier and a support vector machine (SVM) classifier, wherein said chemotherapy response classifier receives an input comprising a marker profile comprising said measurements of said one or more gene products and provides an output comprising said one or more CR scores.

10. The method of claim 9, wherein said chemotherapy response classifier is trained with training data from a plurality of training cancer patients, wherein said training data comprise for each patient of said plurality of training cancer patients (i) a training marker profile comprising measurements of said plurality of gene products in a cell sample taken from said training patient;

and (ii) data indicating whether said training patient is responsive to said treatment regimen.

11. The method of claim 3, comprising determining one or more CR scores that indicates in which percentile said measurements of said one or more gene products fall in the said reference population of individuals, wherein said patient is predicted to be non-responsive if said one or more CR scores indicate that said measurements of said one or more gene products fall in the Y1 percentile in said reference population, wherein Y1 percentile=60 percentile, 70 percentile, 80 percentile, or 90 percentile, or is predicted to be responsive if said one or more CR scores indicate that said measurements of said one or more gene products fall in the Y2 percentile in said reference population, wherein Y2 percentile=10 percentile, 20 percentile, 30 percentile, or 40 percentile,

12. The method of claim 1, wherein said measurements of one or more gene products are measurements of abundance levels of gene transcripts.

13. The method of claim 1, wherein said measurements of one or more gene products are measurements of abundance levels of proteins.

14. The method of claim 1, wherein said chemotherapy regimen comprises administration of a chemotherapy drug selected from the group consisting of 5-fluorouracil, CMF combination consisting of cyclophosphamide, methotrexate, and 5-fluorouracil, paclitaxel, etoposide, and carboplatin.

15. The method of claim 1, wherein said one or more gene products are respectively products of said one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-39.

16. The method of claim 1, wherein said one or more gene products are of at least N or are all of said one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-39, wherein N=2, 3, 4, 5, 10, 15, 20, 25, 30, or 35.

17. The method of claim 1, wherein said one or more gene products are of at least N, or are all of said one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-19. wherein N=2, 3, 4, 5, 10, or 15.

18. The method of claim 1, wherein said one or more gene products are of at least N, or are all of said one or more different genes selected from the group consisting of genes having SEQ ID NOs:

20-39. wherein N=2, 3, 4, 5, 10, or 15.

19. The method of claim 16, wherein said one or more gene products comprises gene products of (i) at least N, or are all of said one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10. or 15 and (ii) at least M, or are all of said one or more different genes selected from the group consisting of genes having SEQ ID NOs;

20-39, wherein M=2, 3, 4, 5, 10, or 15.

20. The method of claim 1, wherein said chemotherapy regimen is an adjuvant chemotherapy regimen, and wherein a prediction of a patient as responsive to said chemotherapy regimen indicates non-occurrence of metastases or survival within a first predetermined period of time after initial diagnosis in said patient treated with said chemotherapy regimen, and wherein a prediction of a patient as non-responsive to said chemotherapy regimen indicates occurrence of metastases or non-survival within a second predetermined period of time in said patient treated with said chemotherapy regimen.

21. The method of claim 1, wherein said chemotherapy regimen is a primary chemotherapy regimen, and a prediction of a patient as responsive to said chemotherapy regimen indicates (i) a reduction in tumor size or number of cancer cells and/or (ii) non-occurrence of metastases or survival within a first predetermined period of time after initial diagnosis in said patient treated with said chemotherapy regimen, and wherein a prediction as responsive to said chemotherapy regimen indicates (iii) a lack of reduction in tumor size or number of cancer cells and/or (iv) occurrence of metastases or non-survival within a second predetermined period of time in said patient treated with said chemotherapy regimen.

22. The method of claim 20 or 21, wherein said first period of time and said second periods of time are the same, and are each 3, 5, 7, 10, or 12 years.

23. The method of claim 1, wherein said patient has been determined to have a poor prognosis, wherein a poor prognosis indicates occurrence of metastases or non-survival within a third predetermined period of time in said patient untreated with any chemotherapy for said cancer.

24. The method of claim 1, wherein said measurement of each said gene product is a relative eve of said gene product in said cell sample versus level of said gene product in a reference sample, represented as a log ratio.

25. The method of claim 24, wherein said reference sample is selected from the group consisting of a sample comprising a pool of cancer cells obtained from a plurality of patients having said cancer, a sample of cells of a non-cancerous cell line of cells of the same type of tissue as said cancer, and a sample of cells of a cell line of said cancer.

26. The method of claim 1, wherein said patient is a human patient.

27. The method of claim 1, wherein said cancer is breast cancer.

28. The method of claim 1, wherein said cancer is ovarian cancer.

29. A method for assigning a treatment regimen for a patient having a cancer, comprising

(i) predicting whether said patient is responsive or non-responsive to a chemotherapy regimen using the method of claim 1;

and

(ii) if said patient is determined to be responsive to said chemotherapy regimen, assigning said patient a treatment regimen that comprises said chemotherapy regimen;

or if said patient is determined to be non-responsive to said chemotherapy regimen, assigning said patient (ii1) a treatment regimen that does not comprise said chemotherapy regimen or (ii2) a treatment regiment comprising (A) said chemotherapy regimen and (B) one or more agents that reduce the expression and/or activity level of said one more gene products.

30. A method for enrolling a plurality of cancer patients for a clinical trial a chemotherapy regimen, comprising (i) determining whether each patient in said plurality is responsive or non-responsive to said chemotherapy regimen using the method of claim 1;

and (ii) assigning each patient who is predicted to be responsive to one patient group and each patient who is predicted to be non-responsive to another patient group, at least one of said patient group being enrolled in said clinical trial.

31. The method of claim 1, wherein said method is a computer implemented method.

32. A computer system comprising

a processor, and

a memory coupled to said processor and encoding one or more programs,

wherein said one or more programs cause the processor to carry out the method of claim 1.

33. A computer program product for use in conjunction with a computer having a processor and a memory connected to the processor, said computer program product comprising a computer readable storage medium having a computer program mechanism encoded thereon, wherein said computer program mechanism may be loaded into the memory of said computer and cause said computer to carry out the method of claim 1.

34. The method of claim 1, further comprising obtaining said measurements of said one or more gene products by a method comprising measuring said plurality of gene products of said cell sample taken from said patient.

35. The method of claim 11, further comprising obtaining measurement of abundance level of each said gene transcript by a method comprising contacting a positionally-addressable microarray with nucleic acids from said cell sample or nucleic acids derived therefrom under hybridization conditions, and detecting the amount of hybridization that occurs, said microarray comprising one or more polynucleotide probes complementary to a hybridizable sequence of each said gene transcript or a nucleic acid derived thereof.

36. The method of claim 11, further comprising obtaining measurement of abundance level of each said gene transcript by a method comprising measuring the transcript level of said gene using quantitative reverse transcriptase PCT (qRT-PCR).

84. The method of claim 34, wherein said method is carried out in vivo.

85. The method of claim 34, wherein said method is carried out in vitro.

37. A method for treating a patient having a cancer, comprising administering to said patient (a) one or more agents that is capable of reducing the expression and/or activity of one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-39 or respective functional equivalents thereof and/or their encoded proteins, and (b) a chemotherapy regimen, wherein said patient is predicted to be non-responsive to said chemotherapy regimen as a result of overexpression of said one or more different genes.

38. The method of claim 37, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-39, wherein N=2, 3, 4, 5, 10, 15, 20, 25, 30, or 35.

39. The method of claim 38, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10, or 15.

40. The method of claim 38, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

20-39, wherein N=2, 3, 4, 5, 10 or 15.

41. The method of claim 38, wherein said one or more different gene are of (i) at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10, or 15; and

(ii) at least K or all of the different genes selected from the group consisting of genes having SEQ ID NOs;

20-39, wherein K=2, 3, 4, 5, 10, or 15.

42. The method of claim 37, wherein said one or more agents comprise a substance selected from the group consisting of siRNA, antis oleic acid, ribozyme, and triple helix forming nucleic acid, each being capable of reducing the expression of one or more c f said one or more different genes.

43. The method of claim 37, wherein said one or more agents comprise a substance selected from the group consisting of antibody, peptide, and small molecule, each is capable of reducing the activity of one or more of proteins encoded by said one or more different genes.

44. The method of claim 42, wherein said one or more agents comprise an siRNA targeting said one or more different genes.

45. The method of claim 44, wherein said one or more different genes consist of at least L different genes, wherein L=2, 3, 4, 5, 10, or 15.

46. The method of claim 37, further comprising determining a transcript level of each of said one or more different genes.

47. The method of claim 46, wherein said determining each said transcript level is carried out by a method comprising measuring the transcript level of said gene using one or more polynucleotide probes, each of said one or more polynucleotide probes comprising a nucleotide sequence complementary to a hybridizable sequence in said transcript of said gene or a nucleic acid derived thereof.

48. The method of claim 47, wherein said one or more polynucleotide probes are polynucleotide probes on a microarray.

49. The method of claim 48, wherein said determining each said transcript level is carried out by a method comprising measuring the transcript level of said gene using quantitative reverse transcriptase PCT (qRT-PCR).

50. The method of claim 37, wherein said chemotherapy regimen comprises administering a chemotherapy drug selected from the group consisting of 5-fluorouracil, CMF combination consisting of cyclophosphamide, methotrexate, and 5-fluorouracil, paclitaxel, etoposide, carboplatin.

51. The method of claim 37, wherein said patient is a human patient.

52. The method of claim 37, wherein said cancer is breast cancer.

53. The method of claim 37, wherein said cancer is ovarian cancer.

54. A method for modulating sensitivity of a cell to a chemotherapeutic drug, comprising contacting said cell with one or more agents, said one or more agents being capable of reducing the expression and/or activity of one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-39 or respective functional equivalents thereof and/or the their encoded proteins.

56. The method of any one of claims 54 to 55, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-39, wherein N=2, 3, 4, 5, 10, 15, 20, 25, 30, or 35.

57. The method of claim 56, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10, or 15.

58. The method of claim 56, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

20-39, wherein N=2, 3, 4, 5, 10, or 15.

59. The method of claim 56, wherein said one or more different gene are of (i) at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10, or 15; and

(ii) at least K or all of the different genes selected from the group consisting of genes having SEQ ID NOs;

20-39, wherein K=2, 3, 4, 5, 10, or 15.

60. The method of claim 54 or 55, wherein said one or more agents comprise a substance selected from the group consisting of siRNA, antisense nucleic acid, ribozyme, and triple helix forming nucleic acid, each is capable of reducing the expression of one or more of said one or more different genes.

61. The method of claim 54 or 55, wherein said one or more agents comprise a substance selected from the group consisting of antibody, peptide, and small molecule, each is capable of reducing the activity of one or more of proteins encoded by said one or more different genes.

62. The method of claim 60, wherein said one or more agents comprise an siRNA targeting said one or more different genes.

63. The method of claim 62, wherein said one or more different genes consist of at least L different genes, wherein L=2, 3, 4, 5, 10, or 15.

64. The method of claim 54 or 55, further comprising determining a transcript level of each of said one or more different genes.

65. The method of claim 64, wherein said determining each said transcript level is carried out by a method comprising measuring the transcript level of said gene using one or more poly:

nucleotide probes, each of said one or more polynucleotide probes comprising a nucleotide sequence complementary to a hybridizable sequence in said transcript of said gene or a nucleic acid derived thereof.

66. The method of claim 65, wherein said one or more polynucleotide probes are polynucleotide probes on a microarray.

67. The method of claim 65, wherein said determining each said transcript level is carried out by a method comprising measuring the transcript level of said gene using quantitative reverse transcriptase PCT (qRT-PCR).

68. The method of claim 54 or 55, wherein said chemotherapeutic drug is selected from the group consisting of 5-fluorouracil, CMF combination consisting of cyclophosphamide, methotrexate, and 5-fluorouracil, paclitaxel, etoposide, carboplatin.

69. The method of claim 54 or 55, wherein said cell is a human cell.

70. The method of claim 54 or 55, wherein said cell is a breast cancer cell.

71. The method of claim 54 or 55, wherein said cell is ovarian cancer cell.

55. A method for modulating growth of a cell, comprising contacting said cell with (a) one or more agents, said one or more agents being capable of reducing the expression and/or activity of one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-39 or respective functional equivalents thereof and/or the their encoded proteins; and

(b) a sufficient amount of a chemotherapeutic drug.

72. A method of identifying an agent that is capable of modulating sensitivity of a cell to the growth inhibitory effect of a chemotherapeutic drug, said method comprising comparing a first growth inhibitory effect of said chemotherapeutic drug on cells expressing said gene in the presence of a candidate agent with a second growth inhibitory effect of said chemotherapeutic drug on cells expressing said gene in the absence of said agent, wherein said agent is capable of reducing the expression and/or activity of a gene selected from the group consisting of genes having SEQ ID NOs:

1-39 or respective functional equivalents thereof and/or its encoded protein, wherein a difference in said first inhibitory effect and said second growth inhibitory effect identities said agent as capable of modulating sensitivity of said cell to the growth inhibitory effect of said chemotherapeutic drug.

73. The method of claim 72, further comprising:

(a) contacting a first cell expressing said gene with said chemotherapeutic drug in the presence of said agent and measuring said first growth inhibitory effect;

(b) contacting a second cell expressing said gene with said chemotherapeutic drug in the absence of said agent and measuring said second growth inhibitory effect.

74. The method of claim 72, wherein said agent comprises a substance selected from the group consisting of siRNA, antisense nucleic acid, ribozyme, and triple helix forming nucleic acid, each reducing the expression of said genes.

75. The method of claim 72, wherein said one or more agents comprise a substance selected from the group consisting of antibody, peptide, and small molecule, each reducing the activity of one or more of proteins encoded by said one or more different genes in said patient.

76. The method of claim 72, wherein said chemotherapy regimen comprises administering a chemotherapy drug selected from the group consisting of 5-fluorouracil, CMF combination consisting of cyclophosphamide, methotrexate, and 5-fluorouracil, paclitaxel, etoposide, carboplatin.

77. The method of claim 72, wherein said cell is a breast cancer cell.

78. The method of claim 72, wherein said cell is ovarian cancer cell.

79. A microarray comprising for each of one or more different genes selected from the group consisting of genes having SEQ ID NOs:

1-39 or respective functional equivalents thereof, one or more polynucleotide probes complementary and hybridizable to a sequence in said gene, wherein polynucleotide probes complementary and hybridizable to said genes constitute at least 50%, 60%, 70%, 80% or 90% of the probes on said microarray.

80. The method of claim 79, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-39, wherein N=2, 3, 4, 5, 10, 15, 20, 25, 30, or 35.

81. The method of claim 79, wherein said one or more different genes consist of at least N or all of the genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10, 15.

82. The method of claim 79, wherein said one or more different genes consist of at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs:

20-39, wherein N=2, 3, 4, 5, 10, or 15.

83. The method of claim 79, wherein said one or more different gene are of (i) at least Nor all of the different genes selected from the group consisting of genes having SEQ ID NOs:

1-19, wherein N=2, 3, 4, 5, 10, or 15; and

(ii) at least N or all of the different genes selected from the group consisting of genes having SEQ ID NOs;

20-39, wherein N=2, 3, 4, 5, 10, or 15.