Coreceptor Tropism Assays

CORECEPTOR TROPISM ASSAYS

Trofile® and Trofile® DNA provide critical information for informed treatment decisions

HIV-1 can attach to human cells either by using the CCR5 coreceptor, the CXCR4 coreceptor, or both (dual/mixed). Tropism testing determines how the virus can attach to the cells in a given patient. Possible tropism results are R5, DM, X4, and X4 near the limit of detection (NLOD).

Why does my patient’s tropism matter? Trofile® DNA HIV tropism results can help you develop a personalized Applies the proven performance of Trofile to cell-associated treatment plan for your patient. Appropriate use of CCR5 viral DNA antagonists including maraviroc requires that an HIV tropism Consider Trofile® DNA when a patient’s viral load is test be performed before initiation of therapy.1 undetectable, tropism is unknown, and substitution with a CCR5 antagonist-containing regimen is desired. Regimen substitution may be considered when2 Trofile® • Laboratory results or clinical adverse events necessitate A highly sensitive assay that provides critical information when a change7,8 selecting a treatment regimen containing maraviroc • Patient exhibits intolerance to the current regimen7 • Trofile was utilized to identify treatment candidates in the • There is concern regarding the long-term effects of the maraviroc multicenter clinical trials.1 current regimen8 • Both the current DHHS and IDSA guidelines recommend tropism testing before initiation of treatment with a CCR5 antagonist.2,3 GenoSure Archive® Plus Trofile® DNA • Trofile is the only commercially available tropism assay Comprehensive suppression management profile that has been clinically validated through use in Phase 2 Designed to provide a comprehensive assessment of and Phase 3 clinical studies to identify CCR5 five antiretroviral drug classes (GenoSure Archive: NRTIs, antagonist candidates.4-6 NNRTIs, PIs, INIs and Trofile DNA: CCR5 antagonist) to facilitate regimen simplification or switches in the setting of Virologic suppression. Monogram Biosciences and LabCorp are pleased to offer options in HIV-1 coreceptor tropism testing.

Samuel H. Pepkowitz, MD, Medical Director 345 Oyster Point Blvd South San Francisco, CA 94080 - Tel: (800) 777-0177 Samuel H. Pepkowitz, MD, Medical Director 345 Oyster Point Blvd Patient Initials: DOB Patient ID/Medical Record # Gender Monogram Accession # South San Francisco, CA 94080 - Tel: (800) 777-0177 Patient Name: DOB Patient ID/Medical Record # Gender Monogram Accession # Date Collected Date Received Date Reported Mode Report Status Date Collected Date Received Date Reported Mode Report Status Investigator Specimen ID Referring Physician Reference Lab ID/Order # Comments: HIV-1 Envelope Subtype: B Comments: HIV-1 Envelope Subtype: B

EXAMPLE EXAMPLE For more information on interpreting this report, please visit www.MonogramHIV.com or call Customer Service at 800-777-0177 between For more information on interpreting this report, please visit www.MonogramHIV.com or call Customer Service at 800-777-0177 between the hours of 6:30am to 5:00pm PT Monday through Friday. the hours of 6:30am to 5:00pm PT Monday through Friday. Trofile uses the complete gp160 coding region of the HIV-1 envelope protein ensuring that all of the determinants of tropism are tested. Subtype is determined based on the HIV-1 gp41 envelope region. Trofile is a cell-based approach to determineapatient's HIV co-receptor tropism (or "Tropotype"). Trofile uses the complete gp160 coding region of the HIV-1 envelope protein ensuring that all of the This assay meets the standards for performance characteristics and all other quality control and assurance requirements established by the Clinical Laboratory Improvement Amendments. The results determinants of tropism are tested. In-house validation data indicates that Trofile is sensitive to detect 0.3% CXCR4 using minor variants. Subtype is determined based on the HIV-1 gp41 envelope have been disclosed to you from confidential records protected by law and are not to be disclosed to unauthorized persons. Further disclosure of these results is prohibited without specific consent of the region. This assay meets the standards for performance characteristics and all other quality control and assurance requirements established by the Clinical Laboratory Improvement Amendments. This persons to whom it pertains, or as permitted by law. test is validated for testing specimens with HIV-1 viral loads equal to or above 1000 copies/mL and should be interpreted only on such specimens. The results should not be used as the sole criteria for **Low level CXCR4 results are near the limit of detection of Trofile and may vary on re-test. patient management. The results have been disclosed to you from confidential records protected by law and are not to be disclosed to unauthorized persons. Further disclosure of these results is Page1of 1 prohibited without specific consent of the persons to whom it pertains, or as permitted by law. LabCorp test: 553100 Page1of 1 LabCorp test: 829670

Consider Trofile® when1 Consider Trofile® DNA when9,10 • Patients have a viral load of ≥1000 copies/mL • Patients have an undetectable viral load Assay features include Assay features include • Phenotypic analysis of viral RNA • Phenotypic analysis of cell-associated viral DNA • Analytically and clinically validated to screen patients prior • Assay performance characteristics analytically validated to initiation of maraviroc* Specimen Requirements** Specimen Requirements** 3mL plasma, EDTA or PPT tube, frozen 4mL whole blood, EDTA tube, frozen

For additional test information, including specimen requirements, CPT coding and RUO/IUO status, consult the Online Test Menu at www.LabCorp.com. Coverage support and assistance for Trofile®, Trofile® DNA, and Trofile® Select are available through Gateway at 877-436-6243. * Trofile® was clinically validated in the maraviroc registrational studies. ** When ordered separately/individually Trofile® Select Consider Trofile Select when tropism testing is desired and viral load is unknown. Allows for ordering of Human Immunodeficiency Virus 1 (HIV-1) coreceptor tropism status to be completed using the appropriate assay, Trofile® or Trofile® DNA, based on the results of an initial prescreening viral assessment by the laboratory. The coreceptor tropism status may be used to determine eligibility for CCR5 antagonist therapy such as maraviroc (Selzentry®).

Specimen References Test Name Test Number 1. SELZENTRY® (maraviroc) prescribing information. Pfizer Labs division of Pfizer Requirements Inc., New York, NY. November 2009. 2. US Department of Health and Human Services. Guidelines for the use of 5mL plasma, EDTA tube, antiretroviral agents in HIV-1-infected adults and adolescents. Available at: http:// www.aidsinfo.nih.gov/Guidelines/Default.aspx?MenuItem=Guidelines&Search= Trofile® Select 553355 frozen and 4mL whole On. Accessed May 2014. 3. Aberg, JA, Kaplan JE, Libman H, et al. Primary care guidelines for the blood, EDTA tube, frozen management of persons with human immunodeficiency virus: 2009 update by the HIV Medicine Association of the Infectious Diseases Society of America. Clin GenoSure Archive® Plus 8mL whole blood, EDTA Infect Dis. 2009;49:651-681. 552020 4. Hardy WD, Gulick R, Mayer H, et al. Efficacy and safety of maraviroc in treatment- Trofile® DNA tube, frozen experienced (TE) patients infected with R5 HIV-1: 96-week combined analysis of the MOTIVATE 1 and 2 studies. Poster presented at: Ninth International Congress on Drug Therapy in HIV Infection; November 9-13, 2008; Glasgow, UK. Poster O425. 5. Saag M, Heera J, Goodrich J, et al. Reanalysis of the MERIT study with the enhanced Trofile® assay (MERIT-ES). Program and abstracts of the 48th Annual ICAAC/IDSA 46th Annual Meeting; October 25-28, 2008; Washington, DC. Abstract H-1232a. 6. Cooper DA, Heera J, Goodrich J et al. Maraviroc versus efavirenz, both in combination with zidovudine-lamivudine, for the treatment of antiretroviral-naive subjects with CCR5-tropic HIV-1 infection. J Infect Dis. 2010;201:803-813. 7 7. Towner W, Klein D, Kerrigan HL, Follansbee S, Yu K, Horberg M. Virologic outcomes of changing enfuvirtide to raltegravir in HIV-1 patients well controlled on an enfuvirtide based regimen: 24-week results of the CHEER study. J Acquir Immune Defic Syndr. 2009;41(4):367-373. 8. Eron JJ, Cooper DA, Youle M, et al. Switch to a raltegravir-based regimen versus continuation of a lopinavir-ritonavir-based regimen in stable HIV-infected patients with suppressed viraemia (SWITCHMRK 1 and 2): two multicentre, double-blind, randomised controlled trials. Lancet. 2010 Jan 30;375(9712):396-407. 9. Toma J, Frantzell A, Cook J, et al. Phenotypic determination of HIV-1 coreceptor tropism using cell-associated DNA derived from blood samples. Paper presented at: Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC); September 12-15, 2010; Boston, Mass. 10. Tashima KT, Coakley E, Toma J, et al. A comparison of DNA and plasma Trofile results in a treatment experienced study population prior to initiation of new antiretroviral regimens. Abstract presented at: the International Workshop on SELZENTRY® is a registered trademark of PHIVCO-1 LLC. HIV & Hepatitis Virus Drug Resistance and Curative Strategies; June 7-11, 2011; Los ©2016 Laboratory Corporation of America® Holdings All rights reserved. L9669-0516-6 Cabos, Mexico. Abstract 91.