Data Summary

Prosigna provides highly concordant results between multiple sites and operators1,2

Prosigna’s analytical validation study demonstrated high precision and reproducibility across multiple qualified laboratories2

Objective: assess 2 aspects of Prosigna’s analytical robustness when used in qualified clinical laboratories: reproducibility when starting from tissue and precision when starting from RNA

Starting from tissue, total standard deviation was only 2.9 Risk of Recurrence (ROR) units across 3 independent sites

  • Reproducibility was assessed by testing a panel of 43 FFPE blocks across 3 sites, including independent H&E review, macrodissection, and RNA isolation by lab personnel at each site
  • Average site-to-site concordance of risk category was >90%, and there were no low-to-high-risk misclassifications or vice versa
  • Average site-to-site concordance of subtype classification was 97%

 

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Starting from RNA, total standard deviation was <1 ROR unit regardless of testing site, operator, and reagent lot

  • Precision was assessed by testing 5 pooled FFPE breast tumor RNA samples >100 times each
  • RNA samples were run independently at 3 different testing sites by a total of 6 different operators using 3 different reagent lots
  • The range of ROR scores for 108 independent measurements was ≤4 units for each of the 5 sample pools
  • 100% concordance was shown between measured and expected subtype result and risk group

 

 

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References: 1. Prosigna [Package Insert]. Seattle, WA: NanoString Technologies, Inc; 2013. 2. Nielsen T, Wallden B, Schaper C, et al. Analytical validation of the PAM50-based Prosigna Breast Cancer Prognostic Gene Signature Assay and nCounter Analysis System using formalin-fixed paraffin-embedded breast tumor specimens [published online ahead of print March 13, 2014]. BMC Cancer. doi:10.1186/1471-2407-14-177. 3. Nielsen T, McDonald S, Kulkarni S, et al. Analytical reproducibility of the breast cancer intrinsic subtyping test and nCounter® Analysis System using formalin-fixed paraffin-embedded (FFPE) breast tumor specimens. Poster presented at: 102nd United States and Canadian Academy of Pathology Annual Meeting; March 2-8, 2013; Baltimore, MD. Abstract 2075.

Prosigna precision: validated in 2 clinical studies with more than 2400 patients1

Prosigna clinical validation studies provide Level 1 evidence of clinical validity2

TransATAC and ABCSG-8 trials featured similar study designs and patient populations1

  • The similar study designs enable combined analysis for a cumulative validation pool of >2400 patients
  • Both studies analyzed samples from postmenopausal women with early-stage, hormone receptor–positive breast cancer who received 5 years of endocrine therapy after surgical resection of the primary tumor
  • Studies included a total of 1786 node-negative patients and 688 node-positive patients, encompassing all risk groups and all 4 intrinsic subtypes
  • The results of the validation studies constitute Level 1 evidence for clinical validity of the Prosigna test for predicting the risk of distant recurrence in postmenopausal women with hormone receptor–positive, early-stage breast cancer2

Prosigna provides prognostic information beyond that obtained from clinical and pathological variables1

Risk groups and prognosis

  • Prosigna ROR is significantly correlated to 10-year distant recurrence (P<0.0001), and risk groups have different rates of DRFS in node-negative and node-positive patients
  • The low-risk group had a 10-year risk of distant recurrence of <5%, while the high-risk group had a 10-year risk of distant recurrence of >20% in node-negative patients

 

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Subtypes and prognosis

  • Luminal A and luminal B subtypes have distinct prognostic outcomes, which impact systemic treatment considerations1,3
  • The choice of systemic therapy should follow intrinsic subtype3

 

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References: 1. Prosigna [Package Insert]. Seattle, WA: NanoString Technologies, Inc; 2013. 2. Gnant M, Filipits M, Greil R, et al; Austrian Breast and Colorectal Cancer Study Group. Predicting distant recurrence in receptor-positive breast cancer patients with limited clinicopathological risk: using the PAM50 Risk of Recurrence score in 1478 postmenopausal patients of the ABCSG-8 trial treated with adjuvant endocrine therapy alone. Ann Oncol. 2014;25(2):339-345. 3. Goldhirsch A, Wood WC, Coates AS, et al. Strategies for subtypes—dealing with the diversity of breast cancer: highlights of the St. Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2011. Ann Oncol. 2011;22(8):1736-1747.

Prosigna precision: validated in 2 clinical studies with more than 2400 patients1

Prognostic significance: proven in the TransATAC study

Study consisted of more than 1000 patient samples1

  • Study cohort consists of FFPE breast tumor tissue samples retrospectively collected and archived from the Anastrozole or Tamoxifen Alone or Combined (ATAC) trial1
  • RNA was extracted from 1372 tumor blocks by Genomic Health from postmenopausal patients with hormone receptor–positive primary breast cancer in the monotherapy arms of ATAC. Twenty-one genes were assessed by quantitative reverse transcriptase polymerase chain reaction, and the Oncotype DX® recurrence score (RS) was calculated2,3
  • ATAC RNA samples isolated by Genomic Health were run with Prosigna (PAM50) to validate ROR3

 

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Prosigna delivered key prognostic information1

  • In order to achieve comparable risk groups, the cut points used for Oncotype DX were different than those used by Genomic Health. For each test, risk groups were prospectively defined based on predicted probability of 10-year distant recurrence (low risk <10%, intermediate risk 10% to 20%, high risk >20%)1
  • Prosigna ROR was significantly related to 10-year distant recurrence (P<0.0001)1,3
  • Intrinsic subtypes had significantly different outcomes in endocrine-treated patients1
  • ROR segregated more node-negative patients to the high-risk and fewer to the intermediate-risk groups than Oncotype DX1

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References: 1. Prosigna [Package Insert]. Seattle, WA: NanoString Technologies, Inc; 2013. 2. Dowsett M, Sestak I, Lopez-Knowles E, et al. Comparison of PAM50 risk of recurrence score with Oncotype DX and IHC4 for predicting risk of distant recurrence after endocrine therapy. J Clin Oncol. 2013;31(22):2783-2790. 3. Dowsett M, Cuzick J, Wale C, et al. Prediction of risk of distant recurrence using the 21-gene recurrence score in node-negative and node-positive postmenopausal patients with breast cancer treated with anastrozole or tamoxifen: a TransATAC study. J Clin Oncol. 2010;28(11):1829-1834.

Prosigna precision: validated in 2 clinical studies with more than 2400 patients1

Prognostic significance: proven in the ABCSG-8 study

Study consisted of more than 1400 patient samples1

  • Study cohort consists of FFPE breast tumor tissue samples retrospectively collected and archived in the Austrian Breast & Colorectal Cancer Study Group (ABCSG) tumor bank from patients enrolled in the ABCSG-8 trial1
  • Postmenopausal women with hormone receptor–positive breast cancer were randomized prior to treatment to 2 years of adjuvant tamoxifen followed by either 3 years of anastrozole or 3 years of adjuvant tamoxifen1,2
  • Reconsented >1200 patients
  • Clinical treatment score (CTS) was derived from standard clinical covariates, including age, grade, tumor size, nodal status, and adjuvant therapy1

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ABCSG-8 objectives1

  • Determine whether the ROR score adds prognostic information beyond the CTS
  • Determine whether the ROR-based risk groups of low, intermediate, and high add prognostic information beyond the CTS
  • Determine whether luminal A and luminal B subtypes have significantly different rates of DRFS

Get more prognostic information than standard clinical variables

Intrinsic subtype

  • Prognosis varied by intrinsic subtype1
  • In all nodal status groups, differentiation between luminal A and luminal B added statistically significant prognostic information beyond CTS1

 

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ROR score

  • Both the continuous ROR score and categorical ROR-based risk groups added statistically significant prognostic information beyond the CTS (P<0.0001)1
  • The low-risk group had an average 10-year DRFS of >95%, while the high-risk group had an average 10-year DRFS of <80%3

 

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References: 1. Prosigna [Package Insert]. Seattle, WA: NanoString Technologies, Inc; 2013. 2. Dubsky PC, Jakesz R, Mlineritsch B, et al. Tamoxifen and anastrozole as a sequencing strategy: a randomized controlled trial in postmenopausal patients with endocrine-responsive early breast cancer from the Austrian Breast and Colorectal Cancer Study Group. J Clin Oncol. 2012;30(7):722-728. 3. Gnant M, Filipits M, Greil R, et al; Austrian Breast and Colorectal Cancer Study Group. Predicting distant recurrence in receptor-positive breast cancer patients with limited clinicopathological risk: using the PAM50 Risk of Recurrence score in 1478 postmenopausal patients of the ABCSG-8 trial treated with adjuvant endocrine therapy alone. Ann Oncol. 2014;25(2):339-345.

Prosigna is indicated for use in postmenopausal women with hormone receptor-positive, node-negative (Stage I or II) or node-positive (Stage II or IIIA) early-stage breast cancer to be treated with adjuvant endocrine therapy.

 

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