Companion diagnostics for cancer and screening methods to identify companion diagnostics for cancer based on splicing variants
A method of classifying a patient for eligibility for cancer therapy based on the presence or absence of splicing variants in a sample of the patient's cancer tissue. Also, a method of screening cancer therapies for efficacy against splicing variants. More specifically, the methods relate to novel splicing variants of genes associated with cancer risk and survival, particularly splicing variants of PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1. Also more specifically, the methods relate to classifying a patient for eligibility for cancer therapy involving the use of GS-1101.
1. A method of detecting a Phosphoinositide 3-kinase catalytic delta (PIK3CD) splice variant comprising:
a) obtaining a tissue sample from a patient; and
b) detecting the presence of a PIK3CD transcript without exon 20 in the sample's total mRNA.
2. The method of claim 1 , further comprising extracting the total mRNA prior to detecting the presence of the PIK3CD transcript without exon 20.
3. The method of claim 2 , further comprising reverse transcribing the total mRNA to cDNA.
4. The method of claim 3 , wherein detecting the presence of PIK3CD transcript without exon 20 is by polymerase chain reaction using the cDNA as a template.
5. The method of claim 3 , wherein detecting the presence of PIK3CD transcript without exon 20 is by cDNA sequencing.
6. The method of claim 3 , wherein detecting the presence of PIK3CD transcript without exon 20 is by polymerase chain reaction using the cDNA as a template with primers specific to exon 20 of PIK3CD transcript and/or a junction of PIK3CD transcript without exon 20.
7. The method of claim 1 , wherein detecting the presence of PIK3CD transcript without exon 20 is by hybridization to probes specific to exon 20 of PIK3 CD transcript and/or a junction of PIK3CD transcript without exon 20.
8. The method of claim 1 , wherein detecting the presence of PIK3CD transcript without exon 20 is by detecting the presence of any part of the PIK3CD mRNA and determining the size of the mRNA such that exon 20 can be said to be absent based on the length of the PIK3CD mRNA.
9. A method of detecting a Phosphoinositide 3-kinase catalytic delta (PIK3CD) splice variant comprising:
a) obtaining a tissue sample from a patient; and
b) detecting the presence of a PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11 in the sample's total mRNA.
10. The method of claim 9 , further comprising extracting the total mRNA prior to detecting the presence of the PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11.
11. The method of claim 10 , further comprising reverse transcribing the total mRNA to cDNA.
12. The method of claim 11 , wherein detecting the presence of PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11 is by polymerase chain reaction using the cDNA as a template.
13. The method of claim 11 , wherein detecting the presence of PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11 is by cDNA sequencing.
14. The method of claim 11 , wherein detecting the presence of PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11 is by polymerase chain reaction using the cDNA as a template with primers specific to exon 20 of PIK3CD transcript and/or a junction of PIK3CD transcript without exon 20.
15. The method of claim 9 , wherein detecting the presence of PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11 is by hybridization to probes specific to exon 20 of PIK3CD transcript and/or a junction of PIK3CD transcript without exon 20.
16. The method of claim 9 , wherein detecting the presence of PIK3CD transcript comprising a nucleic acid sequence of SEQ ID No. 11 is by detecting the presence of any part of the PIK3CD mRNA and determining the size of the mRNA such that exon 20 can be said to be absent based on the length of the PIK3CD mRNA.