Systems and methods for phasing mutations in tumors

Abstract

This application relates generally to analyzing mutations in tumors, and more particularly, to systems and methods for phasing mutations in tumors of subjects (e.g., cancer patients). An exemplary method for phasing mutations in a tumor of a subject comprises enumerating, based on tumor DNA and / or RNA sequence reads, a set of unique mutation patterns observed in the plurality of sequence reads; counting the set of unique patterns observed in the sequence reads to calculate a quantity of each of the unique mutation patterns and / or a quantity of each combination of unique mutation pattern and a transcript group; determining mutation pattern probabilities; and inputting the mutation pattern quantities and the mutation pattern probabilities into a statistical model to estimate at least one of a set of haplotype-existence probabilities that each of the haplotypes exists, a set of haplotype prevalences, and a set of haplotype-transcript prevalences.

Description

Abstract This application generally relates to the analysis of mutations in tumors, and more particularly to systems and methods for phasing mutations in the tumors of subjects (e.g., cancer patients). An exemplary method for phasing mutations in the tumors of subjects includes: enumerating a set of unique mutation patterns observed in multiple sequence reads based on tumor DNA and / or RNA sequence reads; counting the set of unique patterns observed in these sequence reads to calculate the number of each unique mutation pattern and / or the number of each combination of a unique mutation pattern with a transcriptome; determining the mutation pattern probability; and inputting the number of mutation patterns and the mutation pattern probability into a statistical model to estimate at least one of the following: the probability of a set of haplotypes being present, i.e., the probability of each haplotype being present, a set of haplotype prevalences, and a set of haplotype-transcriptome prevalences.

Claims

ATTORNEY DOCKET PCT PATENT APPLICATION 146392064440 P37915-WO 74 of 80 CLAIMS What is claimed is:

1. A method for phasing mutations identified in a tumor of a subject, comprising, by one or more computing devices: accessing a plurality of sequence reads derived from tumor cells obtained from the subject, wherein the sequence reads comprise tumor DNA sequence reads and / or tumor RNA sequence reads; enumerating a set of unique mutation patterns observed in the plurality of sequence reads; counting a number of sequence reads that exhibit each unique mutation pattern of the set of unique mutation patterns observed in the sequence reads to calculate a quantity of each of the unique mutation patterns, and / or counting a number of sequence reads that exhibit each combination of a unique mutation pattern of the set of unique mutation patterns and a transcript group from one or more transcript groups associated with a gene to calculate a quantity of each combination of unique mutation pattern and transcript group; determining, for each of the unique mutation patterns, a probability, for each haplotype of a set of haplotypes, that a hypothetical DNA sequence read from the haplotype will exhibit the unique mutation pattern, and / or a probability, for each combination of a haplotype from the set of haplotypes, a transcript from a set of transcripts associated with the gene, and a transcript group from a set of transcript groups associated with the gene, that a hypothetical RNA sequence read from the haplotype and the transcript will exhibit the unique mutation pattern and the transcript group; inputting the unique mutation pattern quantities and / or unique mutation pattern- transcript group quantities, and the unique mutation pattern probabilities and / or unique mutation pattern-transcript group probabilities into a statistical model to estimate at least one of (i) a set of haplotype-existence probabilities that each of the haplotypes exists, (ii) a set of sf-5934688ATTORNEY DOCKET PCT PATENT APPLICATION 146392064440 P37915-WO 75 of 80 haplotype prevalences, and (iii) a set of haplotype-transcript prevalences; and outputting at least one of (i) the set of haplotype-existence probabilities, (ii) the set of haplotype prevalences, and (iii) the set of haplotype-transcript prevalences.

2. The method of claim 1, wherein the estimation of at least one of the set of haplotype- existence probabilities, the set of haplotype prevalences, and the set of haplotype-transcript prevalences comprises: (i) using the statistical model to sample from at least one of a haplotype-existence posterior probability distribution, a haplotype prevalence posterior probability distribution, and a haplotype-transcript prevalence posterior probability distribution for each haplotype of the set; and (ii) calculating point estimates for haplotype-existence probability, haplotype prevalence, and haplotype-transcript prevalence for each haplotype of the set using the samples from the respective posterior probability distributions.

3. The method of claim 1 or claim 2, further comprising: identifying a set of mutant peptide sequences using in silico translations of one or more haplotype sequences and / or haplotype transcripts based on the set of haplotype-existence probabilities, the set of haplotype prevalences, and / or the set of haplotype-transcript prevalences.

4. The method of any one of claims 1 to 3, wherein the one or more haplotype sequences and / or haplotype transcripts are associated with non-zero haplotype-existence probabilities.

5. The method of claim 3 or claim 4, further comprising: selecting one or more mutant peptide sequences from the set of mutant peptide sequences using one or more predetermined criteria comprising a predetermined criterion that applies to the set of haplotype prevalences and / or the set of haplotype-transcript prevalences.

6. The method of claim 5, wherein the one or more predetermined criteria include a predefined haplotype prevalence threshold and / or a predetermined haplotype-transcript threshold.

7. The method of claim 3 or claim 4, further comprising selecting one or more mutant peptide sequences from the set of mutant peptide sequences by ranking of the set of peptide sequences sf-5934688ATTORNEY DOCKET PCT PATENT APPLICATION 146392064440 P37915-WO 76 of 80 based on the set of haplotype prevalences and / or the set of haplotype-transcript prevalences.

8. The method of any one of claims 3 to 7, further comprising: generating, by a machine-learning model, a prediction of a likelihood of presentation in a major histocompatibility complex (MHC) for one or more of the set of mutant peptide sequences and / or a prediction of an immunogenicity for one or more of the set of mutant peptide sequences.

9. The method of any one of claims 1 to 8, wherein accessing the sequence reads further comprises accessing a plurality of normal DNA sequence reads derived from healthy cells obtained from the subject.

10. The method of claim 9, wherein counting the set of unique mutation patterns observed in the plurality of sequence reads further comprises: calculating a quantity of each unique mutation pattern in the normal DNA sequence reads.

11. The method of any one of claims 1 to 10, wherein counting the set of unique mutation patterns observed in the plurality of sequence reads further comprises: calculating a quantity of each unique mutation pattern in the tumor DNA sequence reads; and / or calculating a quantity of each unique mutation pattern and transcript-group combination in the tumor RNA sequence reads.

12. The method of any one of claims 1 to 11, wherein the probability that the hypothetical RNA sequence read from that haplotype and transcript will exhibit the unique mutation pattern and the transcript group is calculated from a haplotype-transcript prevalence, a conditional probability of observing the combination of unique mutation pattern and the transcript group in an RNA sequence read, and a transcript length.

13. The method of claim 12, wherein calculation of the probability that the hypothetical RNA sequence read from that haplotype and transcript will exhibit the unique mutation pattern and the transcript group further comprises accounting for a probability of miscalling tumor RNA sf-5934688ATTORNEY DOCKET PCT PATENT APPLICATION 146392064440 P37915-WO 77 of 80 sequence reads with the unique mutation pattern.

14. The method of any one of claims 1 to 13, wherein the probability that the hypothetical DNA sequence read from that haplotype will exhibit the unique mutation pattern is calculated from a haplotype prevalence and a conditional probability of observing the unique mutation pattern in a DNA sequence read.

15. The method of claim 14, wherein the calculation of the probability that the hypothetical DNA sequence read from that haplotype will exhibit the unique mutation pattern further comprises accounting for a probability of miscalling tumor DNA sequence reads with the unique mutation pattern.

16. The method of any one of claims 1 to 15, wherein calculation of the probability that the hypothetical RNA sequence from that haplotype and transcript will exhibit the unique mutation pattern and the transcript group further comprises accounting for a probability of a given insert length for the RNA sequence read exhibiting the unique mutation pattern and transcript group.

17. The method of any one of claims 1 to 16, wherein calculation of the probability that the hypothetical DNA sequence from that haplotype will exhibit the unique mutation pattern further comprises accounting for a probability of a given insert length for the DNA sequence read exhibiting the unique mutation pattern.

18. The method of any one of claims 1 to 17, wherein calculation of the probability that the hypothetical RNA sequence from that haplotype and transcript will exhibit the unique mutation pattern and the transcript group further comprises accounting for an RNA sequencing error probability.

19. The method of any one of claims 1 to 18, wherein calculation of the probability that the hypothetical DNA sequence from that haplotype and transcript will exhibit the unique mutation pattern and the transcript group further comprises accounting for a DNA sequencing error probability.

20. The method of any one of claims 1 to 19, wherein estimating at least one of the set of haplotype-existence probabilities, the set of haplotype prevalences, and the set of haplotype- transcript prevalences comprises, for each haplotype of a set of possible haplotypes, using the sf-5934688ATTORNEY DOCKET PCT PATENT APPLICATION 146392064440 P37915-WO 78 of 80 statistical model to: sample a posterior probability distribution for haplotype existence to determine a point estimate for haplotype existence; sample a posterior probability distribution for haplotype prevalence to determine a point estimate for haplotype prevalence; and / or sample a posterior probability distribution for haplotype-transcript prevalence to determine a point estimate for haplotype-transcript prevalence.

21. The method of claim 20, wherein the point estimate for each posterior probability distribution comprises a mean.

22. The method of claim 20 or claim 21, wherein the posterior probability distribution for haplotype existence, haplotype prevalence, and / or haplotype-transcript prevalence is further used to determine an uncertainty associated with haplotype existence, haplotype prevalence, and / or haplotype-transcript prevalence, respectively.

23. The method of any one of claims 1 to 22, wherein accessing the plurality of sequence reads further comprises accessing a set of germline variant and somatic mutation calls derived from tumor cells obtained from the subject.

24. The method of any one of claims 1 to 23, where the statistical model comprises a probabilistic graphical model.

25. The method of any one of claims 1 to 24, wherein the statistical model comprises a hierarchical Bayesian model.

26. The method of claim 25, wherein the hierarchical Bayesian model comprises a haplotype generation model, a DNA Dirichlet-Multinomial model, and / or an RNA Dirichlet-Multinomial model.

27. A system including one or more computing devices, comprising: one or more non-transitory computer-readable storage media including instructions; and one or more processors coupled to the one or more storage media, the one or more sf-5934688ATTORNEY DOCKET PCT PATENT APPLICATION 146392064440 P37915-WO 79 of 80 processors configured to execute the instructions to perform any one of the methods of claims 1 to 26.

28. A non-transitory computer-readable medium comprising instructions that, when executed by one or more processors of one or more computing devices, cause the one or more processors to perform any one of the methods of claims 1 to 26.

29. A vaccine comprising: one or more peptides; a plurality of nucleic acids that encode the one or more peptides; or a plurality of cells expressing the one or more peptides, wherein the one or more peptides are selected from among a set of peptides by performing any one of the methods of claims 1 to 26.

30. A method of manufacturing a vaccine comprising: performing the method of any one of claims 1 to 26 to select one or more peptides from among a set of peptides; and producing a vaccine comprising: at least one of the one or more selected peptides; a plurality of nucleic acids that encode at least one of the one or more selected peptides; or a plurality of cells expressing at least one of the one or more peptides.

31. A pharmaceutical composition comprising one or more peptides selected from among a set of peptides by performing any one of the methods of claims 1 to 26. sf-5934688

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