Method to determine responsiveness of cancer to epidermal growth factor receptor targeting treatments
View Patent ↗The present invention is directed to a method for determining the responsiveness of cancer to an epidermal growth factor receptor (EGFR) treatment. In a preferred embodiment, the presence of at least one variance in the kinase domain of the erbB1 gene confers sensitivity to the tyrosine kinase inhibitor gefitinib. Thus, a diagnostic assay for these mutations will allow for the administration of gefitinib, erlotinib and other tyrosine kinase inhibitors to those patients most likely to respond to the drug.
1. A method for determining an increased likelihood of pharmacological effectiveness of treatment by gefitinib or erlotinib in an individual diagnosed with lung cancer comprising, determining in lung tumor tissue from the individual, the presence or absence of at least one nucleotide variance in exon 18, 19 or 21 of the epidermal growth factor receptor (EGFR) gene, wherein the presence of at least one nucleotide variance selected from:
a. a nucleotide variance in exon 18 comprising a substitution of thymine for guanine or adenine for guanine at nucleotide 2400 of SEQ ID NO: 511, or a substitution of cytosine for guanine at nucleotide 2401 of SEQ ID NO: 511, or resulting in a substitution of Cysteine, Serine or Alanine for Glycine, at amino acid 719 of SEQ ID NO: 512;
b. a nucleotide variance in exon 19 comprising a deletion of nucleotides 2486-2493 of SEQ ID NO: 511, or resulting in a deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747, 748, and 749 of SEQ ID NO: 512; or
c. a nucleotide variance in exon 21 comprising a substitution of guanine for a thymine at nucleotide 2818 of SEQ ID NO: 511, or a substitution of adenine for thymine at nucleotide 2827 of SEQ ID NO: 511, or resulting in a substitution of Arginine for Leucine at amino acid 858 of SEQ ID NO: 512 or resulting in a substitution of Glutamine for Leucine at amino acid 861 of SEQ ID NO: 512;
indicates an increased likelihood of pharmacological effectiveness of treatment by gefitinib or erlotinib in the individual.
2. The method of claim 1 , wherein the nucleotide variance is in exon 18.
3. The method of claim 1 , wherein the nucleotide variance is in exon 19.
4. The method of claim 1 , wherein the nucleotide variance is in exon 21.
5. The method of claim 1 wherein the at least one nucleotide variance is in exon 18 and comprises a substitution of thymine for guanine at nucleotide 2400 of SEQ ID NO: 511.
6. The method of claim 1 , wherein the at least one nucleotide variance is in exon 18 and results in a substitution of Cysteine for Glycine at amino acid 719 of SEQ ID NO: 512.
7. The method of claim 1 , wherein the at least one nucleotide variance is in exon 18 and comprises a substitution of adenine for guanine at nucleotide 2400 of SEQ ID NO: 511.
8. The method of claim 1 , wherein the at least one nucleotide variance is in exon 18 and results in a substitution of Serine for Glycine at amino acid 719 of SEQ ID NO: 512.
9. The method of claim 1 , wherein the at least one nucleotide variance is in exon 18 and comprises a substitution of cytosine for guanine at nucleotide 2401 of SEQ ID NO: 511.
10. The method of claim 1 , wherein the at least one nucleotide variance is in exon 18 and results in a substitution of Alanine for Glycine at amino acid 719 of SEQ ID NO: 512.
11. The method of claim 1 , wherein the at least one nucleotide variance is in exon 19 and comprises a deletion of nucleotides 2486-2493 of SEQ ID NO: 511.
12. The method of claim 1 , wherein the nucleotide variance in exon 19 is selected from the group consisting of a deletion of nucleotides 2481 to 2495, a deletion of nucleotides 2482 to 2496, a deletion of nucleotides 2485-2493, a deletion of nucleotides 2486-2496, a deletion of nucleotides 2486 to 2497, and a deletion of nucleotides 2486 to 2503 of SEQ ID NO: 511.
13. The method of claim 12 , wherein the at least one nucleotide variance is in exon 19 and comprises a deletion of nucleotides 2481 to 2495 of SEQ ID NO: 511.
14. The method of claim 12 , wherein the at least one nucleotide variance is in exon 19 and comprises a deletion of nucleotides 2482 to 2496 of SEQ NO: 511.
15. The method of claim 1 , wherein the at least one nucleotide variance is in exon 19 and comprises a deletion of nucleotides 2486 to 2497 of SEQ ID NO: 511.
16. The method of claim 12 , wherein the at least one nucleotide variance is in exon 19 and comprises a deletion of nucleotides 2486 to 2503 of SEQ ID NO: 511.
17. The method of claim 1 , wherein the at least one nucleotide variance is in exon 19 and results in a deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747, 748 and 749, of SEQ ID NO: 512.
18. The method of claim 17 , wherein the deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747, 748, and 749 of SEQ ID NO: 512, further includes a deletion of amino acid alanine at codon 750.
19. The method of claim 18 , wherein the deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747,748, and 749 of SEQ ID NO: 512, further includes a deletion of amino acid threonine at codon 751.
20. The method of claim 18 , wherein the deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747,748, and 749 of SEQ ID NO: 512, further includes a deletion of amino acid serine at codon 752.
21. The method of claim 1 , wherein the deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747, 748, and 749 of SEQ ID NO: 512, further includes a deletion of amino acid glutamic acid at codon 746 of SEQ ID NO: 512.
22. The method of claim 1 , wherein the nucleotide variance in exon 19 is selected from the group consisting of a deletion of nucleotides 2485 to 2496, a deletion of nucleotides 2483-2501, a deletion of nucleotides 2485-2494, a deletion of nucleotides 2485-2497, a deletion of nucleotides 2484-2501 of SEQ ID NO: 511.
23. The method of claim 1 , wherein the at least one nucleotide variance is in exon 21 and results in a substitution of Arginine for Leucine at amino acid 858 of SEQ ID NO: 512.
24. The method of claim 1 , wherein the at least one nucleotide variance is in exon 21 and comprises a substitution of guanine for thymine at nucleotide 2818 of SEQ ID NO: 511.
25. The method of claim 1 , wherein the at least one nucleotide variance is in exon 21 and comprises a substitution of adenine for thymine at nucleotide 2827 of SEQ ID NO: 511.
26. The method of claim 1 , wherein the at least one nucleotide variance is in exon 21 and results in a substitution of glutamine for leucine at amino acid 861 of SEQ ID NO: 512.
27. The method of claim 1 , wherein the presence or absence of the at least one nucleotide variance is determined by sequencing.
28. The method of claim 1 , wherein the presence or absence of the at least one nucleotide variance is determined by amplification.
29. The method of claim 1 , wherein the presence or absence of the at least one nucleotide variance is determined by single strand conformation polymorphism, denaturing gradient gel electrophoresis or temperature gradient gel electrophoresis analysis.
30. The method of claim 1 , wherein the presence or absence of the at least one nucleotide variance is determined by mismatch cleavage analysis.
31. The method of claim 1 , further comprising determining in a control sample the presence or absence of at least one nucleotide variance in exon 18, 19 or 21 of the epidermal growth factor receptor (EGFR) gene, selected from:
a. a nucleotide variance in exon 18 comprising a substitution of thymine for guanine or adenine for guanine at nucleotide 2400 of SEQ ID NO: 511, or a substitution of cytosine for guanine at nucleotide 2401 of SEQ ID NO: 511, or resulting in a substitution of Cysteine, Serine or Alanine for Glycine, at amino acid 719 of SEQ ID NO: 512;
b. a nucleotide variance in exon 19 comprising a deletion of nucleotides 2486-2493, or resulting in a deletion of at least amino acids leucine, arginine and glutamic acid, at codons 747, 748, and 749 of SEQ ID NO: 512; or
c. a nucleotide variance in exon 21 comprising a substitution of guanine for a thymine at nucleotide 2818 of SEQ ID NO: 511, or a substitution of adenine for thymine at nucleotide 2827 of SEQ ID NO: 511, or resulting in a substitution of Arginine for Leucine at amino acid 858 of SEQ ID NO: 512 or resulting in a substitution of Glutamine for Leucine at amino acid 861 of SEQ ID NO: 512;
and comparing the determined presence or absence in the control sample to the determined presence or absence in the lung tumor tissue.