NANO46 Genes and Methods to Predict Breast Cancer Outcome
The present invention provides methods for classifying and for evaluating the prognosis of a subject having breast cancer are provided. The methods include prediction of breast cancer subtype using a supervised algorithm trained to stratify subjects on the basis of breast cancer intrinsic subtype. The prediction model is based on the gene expression profile of the intrinsic genes listed in Table 1. Further provided are compositions and methods for predicting outcome or response to therapy of a subject diagnosed with or suspected of having breast cancer. These methods are useful for guiding or determining treatment options for a subject afflicted with breast cancer. Methods of the invention further include means for evaluating gene expression profiles, including microarrays and quantitative polymerase chain reaction assays, as well as kits comprising reagents for practicing the methods of the invention.
1 . A method of predicting outcome in a subject having breast cancer comprising:
providing a tumor sample from the subject;
determining the expression of at least the genes in the NANO46 intrinsic gene list of Table 1 in the tumor sample;
determining the intrinsic subtype of the tumor sample, wherein the intrinsic subtype is selected from the group consisting of at least Basal-like, Luminal A, Luminal B or HER2-enriched;
determining a proliferation score based on the expression of a subset of proliferation genes in the NANO46 intrinsic gene list;
calculating a risk of recurrence score using a weighted sum of said intrinsic subtype, proliferation score and optionally one or more clinicopathological variables such as tumor size, nodal status or histological grade; and
determining whether the subject has a low or high risk of recurrence based on the risk of recurrence score.
2 . The method of claim 1 , wherein determining a proliferation signature based on the expression of a subset of proliferation genes in the NANO46 intrinsic gene list comprises determining the expression of each of the NANO46 intrinsic genes selected from ANLN, CCNE1, CDC20, CDC6, CDCA1, CENPF, CEP55, EXO1, KIF2C, KNTC2, MELK, MKI67, ORC6L, PTTG1, RRM2, TYMS, UBE2C and UBE2T.
3 . The method of claim 1 , further comprising determining at least one of the following: tumor grade, tumor ploidy, nodal status, estrogen receptor expression, progesterone receptor expression, and HER2/ERBB2 expression
4 . The method of claim 1 , further comprising determining each of the following: tumor grade, tumor ploidy, nodal status, estrogen receptor expression, progesterone receptor expression, and HER2/ERBB2 expression
5 . The method of claim 1 , wherein the risk of recurrence score is calculated using the following equation:
ROR-PT=−0.0067*Basal+0.4317*Her2+−0.3172*LumA+0.4894*LumB+0.1981*ProliferationScore+0.1133*Tumor Size.
6 . The method of claim 1 , wherein the outcome is breast cancer specific survival, event-free survival or response to therapy.
7 . The method of claim 1 , wherein the expression of the members of the NANO46 intrinsic gene list is determined using the nanoreporter code system (nCounter® Analysis system).
8 . A kit comprising a plurality of probes for determining the expression of at least the genes in the NANO46 intrinsic gene list of Table 1 in a tumor sample for use in a method of predicting outcome in a subject having breast cancer.
9 . The kit of claim 8 , wherein the kit comprises a plurality of probes of Table 1A.
10 . The kit of claim 9 , wherein the kit comprises each of the probes of Table 1A.
11 . The kit of claim 8 , comprising probes for determining the expression of each of the NANO46 intrinsic genes selected from ANLN, CCNE1, CDC20, CDC6, CDCA1, CENPF, CEP55, EXO1, KIF2C, KNTC2, MELK, MKI67, ORC6L, PTTG1, RRM2, TYMS, UBE2C and UBE2T.
12 . The kit of claim 8 , wherein each probe in the plurality of probes comprises a target specific sequence capable of hybridizing to no more than one NANO46 intrinsic gene listed in Table 1, and optionally comprises at least two label attachment regions, said label attachment regions comprising one or more label monomers that emit light.
13 . The kit of claim 9 , wherein the plurality of probes comprises a probe pair to detect the NANO46 intrinsic genes listed in Table 1, wherein each probe in the probe pair comprises a target specific sequence capable of hybridizing to no more than one NANO46 intrinsic gene listed in Table 1 and wherein the target specific sequences bind to different regions of the same NANO46 intrinsic gene.
14 . The kit of claim 13 , wherein one probe of the probe pair further comprises at least two label attachment regions, said label attachment regions comprising one or more label monomers that emit light
15 . The kit of claim 8 , further comprising one or more reagents for determining one or more clinicopathological variables of the tumor sample such as tumor size, tumor grade, tumor ploidy, nodal status, estrogen receptor expression, progesterone receptor expression, and HER2/ERBB2 expression.