K Number
K070675
Device Name
MAMMAPRINT
Manufacturer
Date Cleared
2007-06-22

(102 days)

Product Code
Regulation Number
866.6040
Reference & Predicate Devices
Predicate For
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

MammaPrint® is a qualitative in vitro diagnostic test service, performed in a single laboratory, using the gene expression profile of fresh breast cancer tissue samples to assess a patients' risk for distant metastasis.

The test is performed for breast cancer patients who are less than 61 years old, with Stage I or Stage II disease, with tumor size ≤ 5.0 cm and lymph node negative. The MammaPrint® result is indicated for use by physicians as a prognostic marker only, along with other clinicopathological factors.

Device Description

The MammaPrint service is a microarray based gene expression analysis of a tumor. The analysis is based on several processes: using fresh tissue stored in RNAlater, isolation of RNA from frozen tumor tissue sections, DNA'se treatment of isolated RNA, linear amplification and labeling of DNA'se treated RNA, cRNA purification, hybridization of the cRNA to the MammaPrint microarray, scanning the MammaPrint microarray and data acquisition (feature extraction), calculation and determination of the risk of recurrence in breast cancer patients.

The MammaPrint analysis is designed to determine the gene activity of specific genes in a tissue sample compared to a reference standard. The result is an expression profile, or fingerprint, of the sample.

The correlation of the sample expression profile to a template (the mean expression profile of 44 tumors with a known good clinical outcome) is calculated and the molecular profile of the sample is determined (Low Risk, High Risk).

AI/ML Overview

This document describes the MammaPrint® device, a gene expression profiling test system for breast cancer prognosis.

Acceptance Criteria and Device Performance

CriteriaReported Device Performance
Analytical Accuracy98.5% (based on 190+ independent analyses of control samples)
Classification Accuracy97.7% for High Risk/Low Risk classification
Borderline SamplesLess than 5% of analyzed samples
RepeatabilityConsidered Precise, Reproducible, Sensitive, Specific, Accurate, and Robust

Study Details

2. Sample Size and Data Provenance

  • Training Set Sample Size:
    • Nature Paper (1): 78 patients
    • NEJM Paper (2): 151 patients
  • Validation Set Sample Size:
    • NEJM Paper (2): 151 patients (used for both training and validation)
    • Transbig Paper (4): 302 patients
  • Data Provenance: Not explicitly stated, but the studies mention "independent European validation" for the Transbig Paper, implying data from Europe. The "comprehensive three-way inter-laboratory comparison study between three independent laboratories in three different countries (Dutch, French and U.S.)" indicates data from these countries for analytical performance, but not specifically for clinical studies. Retrospective or prospective nature is not specified, but the "time frame" for clinical studies (e.g., 2002, 2006) and the reporting of 5-year and 10-year metastasis-free survival strongly suggest retrospective analysis of existing patient cohorts.

3. Number of Experts and Qualifications for Ground Truth

  • Not explicitly stated for the clinical studies. The clinical performance is based on outcomes data (metastasis-free survival) rather than expert consensus on individual cases.

4. Adjudication Method for Test Set

  • Not applicable as the ground truth is based on clinical outcomes (metastasis-free survival) rather than expert consensus requiring adjudication.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

  • No, a multi-reader multi-case (MRMC) comparative effectiveness study was not performed or mentioned in the provided text. The device is a diagnostic test, not an AI-assisted interpretation tool for human readers.

6. Standalone Performance Study

  • Yes, the performance reported is a standalone (algorithm only) performance. The MammaPrint is a gene expression analysis that directly provides a risk classification (High Risk or Low Risk) based on the gene activity profile. The "Classification performance" states "the accuracy of classifying a sample as High Risk or Low Risk, is 97.7% (i.e., 1.1% false negative classification)," which refers to the algorithm's performance.

7. Type of Ground Truth Used for Clinical Studies

  • Outcomes Data: The clinical studies (Nature Paper, NEJM Paper, Transbig Paper) used clinical outcomes data, specifically metastasis-free survival at 5 and 10 years, as the ground truth for validating the prognostic ability of the MammaPrint profile. This is indicated by phrases like "Metastasis-free survival by profile at 10 yrs."

8. Sample Size for the Training Set

  • As detailed in Section 2, the training set sizes were:
    • Nature Paper (1): 78 patients
    • NEJM Paper (2): 151 patients (also used for validation)

9. How the Ground Truth for the Training Set Was Established

  • The ground truth for the training set (and validation set) was established using clinical outcomes data, specifically the occurrence of distant metastasis over time. The studies measured "metastasis-free survival" in patient cohorts. This implies that the initial patient samples were analyzed by MammaPrint, and then patients were followed over several years to observe their clinical outcome regarding metastasis.

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Section 5: 510(k) Summary

  1. Assigned 510(k) number The assigned 510(k) number is

is K070675

JUN 2 2 2007

  1. Company Agendia BV Slotervaart Medical Center 9D Louwesweg 6, 1066EC Amsterdam The Netherlands : 31 20 512 9161 Telephone : 31 20 51291 62 Facsimile

  2. Contact Guido Brink, Director Quality Management and Regulatory Affairs

  3. Date Prepared March 1st 2007

  4. Proprietary Name MammaPrint®

6. Classification Name

Gene expression profiling test system, for breast cancer prognosis.

7. Common Name

Multivariate device for cancer prognosis

8. Classification

Class II, regulated under 21 CFR 866.6040, product code NYI

  1. Predicate Device

Agendia BV's MammaPrint (K062694)

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10. Device Description

The MammaPrint service is a microarray based gene expression analysis of a tumor. The analysis is based on several processes: using fresh tissue stored in RNAlater, isolation of RNA from frozen tumor tissue sections, DNA'se treatment of isolated RNA, linear amplification and labeling of DNA'se treated RNA, cRNA purification, hybridization of the cRNA to the MammaPrint microarray, scanning the MammaPrint microarray and data acquisition (feature extraction), calculation and determination of the risk of recurrence in breast cancer patients.

The MammaPrint analysis is designed to determine the gene activity of specific genes in a tissue sample compared to a reference standard. The result is an expression profile, or fingerprint, of the sample.

The correlation of the sample expression profile to a template (the mean expression profile of 44 tumors with a known good clinical outcome) is calculated and the molecular profile of the sample is determined (Low Risk, High Risk).

11. Intended Use

MammaPrint is a qualitative in vitro diagnostic test service, performed in a single laboratory, using the gene expression profile of fresh breast cancer tissue samples to assess a patient's risk for distant metastasis.

The test is performed for breast cancer patients who are less than 61 years old, with Stage I or Stage II disease, with tumor size <= 5.0 cm and who are lymph node negative. The MammaPrint result is indicated for use by physicians as a prognostic marker only, along with other clinicopathological factors.

12. Performance Data (non-clinical)

Analytical performance

MammaPrint analytical (i.e., non-clinical) performance characteristics investigated comprise Precision, Reproducibility, Cutoff, Sensitivity、 Specificity, Accuracy, Robustness and Ruggedness.

The technical validity of MammaPrint is determined on multiple individual validation experiments; a comprehensive three-way inter-laboratory comparison study between three independent laboratories in three different countries (Dutch, French and U.S.); data of about 200 analyses of two reference samples over a period of 12 months, used to

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monitor experiment-to-experiment quality; and quality controls for which the cut-off for all QCs is based on over 5000 hybridizations (2500 samples) performed at Agendia.

Comparison validation experiments performed on over 50 human tissue tumor pairs and 6 mice xenografts, preserved using two different tissue preservation methods (fresh frozen and fresh RNAlater preservation at room temperature), show similar analytical results (MammaPrint Index) and prognostic outcome (High Risk)Low Risk) of the MammaPrint device.

Based on 12 month repeated experiments of a Low Risk and High Risk control sample (i.e., more than 190 independent analyses), the Analytical Accuracy of the measurement is 98,5%.

Classification performance

Based on the analytical performance of MammaPrint, the accuracy of classifying a sample as High Risk or Low Risk, is 97.7% (i.e., 1.1% false negative classification).

Borderline Sample

As a result of the technical inaccuracy, analytical measurements (i.e., MammaPrint Index) can fall within a pre-defined area around the classification cut-off between the High Risk and Low Risk profile (i.e., "Borderline Sample").

Based on the analyses results of independent MammaPrint analyses over a time period of over 2 years, it has been shown that less than 5% of the analyzed samples are considered to be "Borderline Samples".

"Borderline Samples" have a less than 90% classification accuracy (i.e. >10% chance of false classification).

Looking at all generated data it is concluded that the MammaPrint analysis is considered to be Precise, Reproducible, Sensitive, Specific, Accurate and Robust and valid for the Intended Use.

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13. Clinical Data

Clinical performance testing is based on the following studies:

StudyPurposeTime FrameComments
Nature Paper (1)Type: TrainingDevelopment ofbreast cancerprognosis 70-geneprofile (LNO, <55)2002, 78 patients,6.4% adjuvanttreatmentWithin 5 yearmetastasis risk byprofile multivariateOR 18
NEJM Paper (2)Type: Training andValidationValidation of the70-gene profile inconsecutive seriesof breast cancerpatients (LNO, <53)2002, 151 patients,5.2% adjuvanttreatmentMetastasis-freesurvival by profile at10 yrs: low riskprofile 87%, highrisk profile 44%(at 5 yrs: 93% and56% respectively)
MammaPrint Paper(3)Type: Training andValidationDevelopment ofMammaPrint2006, reproducibilityof (1) and (2) onMammaPrintHighly reproducibleMammaPrint asdiagnostic tool
Transbig Paper (4)Type: ValidationIndependentEuropeanvalidation of 70-gene signature(LNO, <61)2006, 302 patients,no adjuvanttreatmentMetastasis-freesurvival by profile at10 yrs: low riskprofile 88%, highrisk profile 71%(at 5 yrs: 96% and83% respectively)

14. Conclusion

The MammaPrint device subject of this 510(k) using fresh tissue is substantially equivalent to the original MammaPrint under K062694

(1) Gene expression profiling predicts clinical outcome of breast cancer;

Laura J. Van 't Veer et al; Nature (2002) 415, p530-536.

(2) A Gene-Expression Signature as a Predictor of Survival in Breast Cancer;

Marc J. Van de Vijver et al;. New Engl J Med (2002) 347, p1999-2009.

(3) Converting a breast cancer microarray signature into a high-throughput diagnostic test; Annuska M. Glas et al; BMC Genomics (2006) accepted.

(4) Validation and clinical utility of a 70-gene prognostic signature for women with nodenegative breast cancer; Marc Buyse et al; J Natl Cancer Inst (2006), 98, p1183-1192.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/4/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized caduceus symbol, which is a staff with two snakes entwined around it. The symbol is placed in the center of a circular border. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" is written around the border of the circle.

Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

JUN 2 2 2007

Agendia BV c/o Mr. Guido Brink Director Regulatory Affairs Slotervaart Medical Center. Floor 9D Louwesweg 6, 1066 EC Amsterdam The Netherlands

Re: K070675

Trade/Device Name: MammaPrint® Regulation Number: 21 CFR 866.6040 Regulation Name: Gene expression profiling test system for breast cancer prognosis Regulatory Class: Class II Product Code: NYI Dated: May 31, 2007 Received: June 4, 2007

Dear Mr. Brink:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820). This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The

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Page 2 -

FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (240) 276-3150 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Robert H. Becker Jr.

Robert L. Becker, Jr., M.D., Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known): K070675

Device Name: MammaPrint®

Indications For Use:

MammaPrint® is a qualitative in vitro diagnostic test service, performed in a single laboratory, using the gene expression profile of fresh breast cancer tissue samples to assess a patients' risk for distant metastasis.

The test is performed for breast cancer patients who are less than 61 years old, with Stage I or Stage II disease, with tumor size ≤ 5.0 cm and lymph node negative. The MammaPrint® result is indicated for use by physicians as a prognostic marker only, along with other clinicopathological factors.

Prescription Use (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

જી

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

ia m chan

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K070675

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§ 866.6040 Gene expression profiling test system for breast cancer prognosis.

(a)
Identification. A gene expression profiling test system for breast cancer prognosis is a device that measures the ribonucleic acid (RNA) expression level of multiple genes and combines this information to yield a signature (pattern or classifier or index) to aid in prognosis of previously diagnosed breast cancer.(b)
Classification. Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Gene Expression Profiling Test System for Breast Cancer Prognosis.” See § 866.1(e) for the availability of this guidance document.