The Making of a Translational Researcher

1. The Making of a Translational Researcher Lupe G. Salazar, M.D. Tumor Vaccine Group Division of Oncology

2. In the Beginning…. • Started in hematology with interest in BMT research • Interest dwindled after 4 months of BMT in-pt. service • Panic sets in at the end of 1st clinical year • Meet future mentor on oncology service • Sparks my interest in tumor immunology • Vaccines targeting HER2 in breast/ovarian Ca • Join the Tumor Vaccine Group with specific interest in clinical development of HER2 vaccines in breast and ovarian cancer

3. Going from Big to Small • What does “clinical development of HER2 vaccines in breast and ovarian cancer” mean? • Clinically oriented- I didn’t want to be under a hood or taking care of mice all day • Yet I was very interested in the basic science aspects • Biology of the disease (humans and mouse models) • Tumorigenic and immunogenic aspects of HER2 protein • Formulation of HER2 vaccines • Monitoring of immune responses to HER2 vaccination

4. A Leap of Faith and Lots of Luck • Take a vaccine in pre-clinical development and design a study to test it in humans • DNA plasmid based vaccine encoding the intracellular domain (ICD) of HER2 • Designing the study and deciding what questions to ask • The product itself- first time use in humans • Current literature on DNA vaccines- malaria and HIV • Mentors previous experience with peptide vaccines • Based on the above • Phase I study to look at safety and immunogenicity • Enroll enough subjects to be able to ask scientific questions

5. A Phase I Trial to Evaluate the Safety and Immunogenicity of a DNA Plasmid Based Vaccine Encoding the HER2 Intracellular Domain in Subjects with HER2 Overexpressing Tumors • Primary objectives • To determine the safety of intradermal administration of 3 doses of a DNA based HER2 vaccine administered with a fixed dose of GM-CSF • To determine whether a plasmid DNA HER2 vaccine can elicit HER2 specific immune responses • Secondary objectives • To determine if the dose of the plasmid-based DNA vaccine effects immunologic responses • To determine the persistence of DNA at the site of vaccination.

6. Phase I Study of HER2 ICD DNA Vaccine • Patients with stage III/IV HER2 + tumors • Breast and ovarian • Dose-escalation study: 10µg, 100µg, 1,000µg • Most immunogenic dose • 22 patients/dose • Sufficient to gather immune response data • Vaccinate monthly for 3 months • GM-CSF as an adjuvant • Based on previous vaccine trials and animal data

7. K23 Research Questions • K23 specific aims based on the clinical trial • Evaluate the safety and measure the extent of immunogenicity of HER2 (ICD) plasmid-based DNA vaccination in patients with advanced stage HER2 overexpressing breast and ovarian cancer • Determine whether HER2 specific T cell memory occurs after active immunization with a HER2 ICD plasmid-based DNA vaccine • Determine if prolonged local antigen expression is associated with the development of a HER2 specific memory T cell response after immunization.

8. Methods • Phase I study: 22 subjects at each dose-level • Provide adequate numbers of subjects for gathering (1) safety data, (2) statistically powered immunologic response data and (3) statistically powered data comparing the immunologic efficacy of the 3 doses of vaccine • Immunologic Correlates • Assess generation of immunologic memory • Proliferation and ELIspot assays to determine HER2 specific T cell response • Cytokine Flow cytometry to characterize memory T cells • DTH response post- immunization • Assess prolonged local antigen expression (IHC) • Chronic deposition of CD1a+ cells • in vivo transfection of APC • Chronic expression of HER2 protein

9. Timeline • Protocol Concept- March 2001 • Protocol designed-July 2001 • (AACR/ASCO Clinical Trials Course)- • Pre-IND Meeting with FDA- August 2001 • Pre-clinical testing, manufacturing, vialing of Product, multiple revisions of protocol • K23 submitted June 2002 (Awarded June 2003) • IND Meeting with FDA- July 2003 • Final IND submission-January, 2004 • Approval from FDA/IRB/GCRC April 2004 • Phase I study enrolls 1st subject-May, 2004

10. Some Results • 9 subjects enrolled at lowest dose (10mcg) • 6 Stage IIIB • 3 Stage IV (2 with stable bony disease) • 5 have completed 3 vaccines • Toxicity • Grade I skin reaction at injection site • Grade I Autoimmune: ANA 1:40 • Grade I Myalgias • No evidence of plasmid at vaccination site 1 month post • Immunologic Correlates • 4/5 patients have generated HER2 specific T cell responses after the 2nd vaccine • Skin biopsies currently being evaluated (IHC)

11. Financing a Clinical Trial • It takes lots of money to fund the needed resources • Research Coordinators • Research Nurses • Lab Personnel • NIH/NCI funding • RO1 (Nora Disis) to run the study • K23 pay young investigator salary including travel and few lab supplies and 5% of lab tech • Institutional resources for young investigators • FHCRC RTO • GCRC

12. Things That Can Slow Down the Process • Regulatory Agencies and Committees • FDA • SRC (FHCRC Scientific Committee) • UW HSD • GCRC • Things that might help • Pay attention to detail • Pay attention to deadlines • Quick responses from the PI

13. Acknowledgements Tumor Vaccine Group Jennifer Childs, M.P.H. Yushe Dang, Ph.D. Corazon dela Rosa, BSMT Nora Disis, M.D. Patty Fintak, M.A. Ekram Gad, Ph.D. Vivian Goodell, M.P.H. Heidi Gray, M.D. Doreen Higgins, R.N., B.S.N. Karilynn Howard, M.S. Hailing Lu, Ph.D. Bob Schroeder, B.A. Ron Swenson, M.D. Nate Van Denend, B.S. George Vielhauer, Pharm.D./Ph.D. Wolfgang Wagner, Ph.D. Sarah Wallace, M.S. Devon Webster, M.D. Collaborators Katherine Guthrie, Ph.D., FHCRC Ted Gooley, Ph.D., FHCRC Holden Maeker, Ph.D., BD Biosciences Skip Maino, Ph.D.,BD Biosciences www.tumorvaccinegroup.org