figure 12 ad5f35 human her2/neu
ad5f35-her2-ectm lot vmc1003

Conc: 1E+12 vp/mL Vol: 0.2 mL Store: < = -70C Exp: 12/21/2015 **(stability studies pending)** Made by CAGT Baylor, Houston, Texas New Drug For Investigational Use Only Total: 519 vials There is also a Virus Master Archive for this clinical lot (VMA1003): 1E=12 vp/ml, 1.0mL/vial. Total: 30 vials.  
A second clinical lot of **vector (VEC1003.1.11**) was produced and vialed on 02/09/12. It will be *available for use in future investigational studies* of the Ad5f35HER2ECTM vector as well as in this study should the first lot be used up or become unusable. This second clinical lot is labeled as follows:
Ad5f35-Her2-ECTM Lot: VEC1003.1.11 Conc: 1E+12 vp/mL Volume: 0.2 mL Store: < = -70C Date Fill: 02/09/2012 Made by CAGT Baylor, Houston, Texas New Drug For Investigational Use Only The vialed vector is stored below -70C at the NIH Clinical Center Investigational Drug Management Research Section (IDMRS). Investigational drug supply of vector used for manufacture of autologous HER2 DC vaccines will be tracked and documented as per regulatory requirements.  
13.1.3.2 **Proposed Vector Product Use and Formulation:** Vector-Transduced Autologous Dendritic Cell Vaccine 13.1.3.3 **Product Name:** Autologous Ad5f35HER2ECTM transduced dendritic cell vaccine
(AdHER2 DC Vaccine)
13.1.3.4 Dendritic Cell Vaccine Manufacturing Summary The autologous AdHER2 DC vaccine will be manufactured by the Department of Transfusion Medicine (DTM) at the NIH Clinical Center in Bethesda, MD:
National Institutes of Health NIH Clinical Center Department of Transfusion Medicine 10 Center Drive-MSC 1184 Building 10, Room 1C711 Bethesda, MD 20892-1184 The Sponsor-Investigator held IND that will be submitted for this product will include a Letter of Authorization to Cross-Reference BB-MF 11054 from the Master File Type V for Facility, Operational and Quality Systems for the Manufacture of Cellular Therapy Products that is held by DTM. Refer to Appendix 3 **for the DTM Standard Operating Procedure for Manufacture of the** AdHER2DC vaccine 13.1.4 Stability and Storage  
  
Vector Stability and Release Testing Vector stability and release testing is being conducted at 0 months, 12 months and yearly thereafter as outlined below. Additional testing is conducted for infectious units (IU), viral particles (VP). USP sterility, endotoxin, pH, transgene expression and restriction digest.  
Vialed vector is stored at < -70 degrees Celsius. 13.1.5 Administration Procedures Refer to **Appendix 3** for details regarding Ad5f35HER2ECTM vector use in the transduction of autologous dendritic cells and generation of final vaccine product. 13.1.6 Incompatibilities The Ad5f35HER2ECTM vector has no known incompatibilities in culture.
figure 12 ad5f35 human her2/neu
13.2 sargramostim 13.2.1 source

The GM-CSF to be used in this study is glycosylated, recombinant human GranulocyteMacrophage Colony Stimulating Factor (rhu GM-CSF) (brand name Leukine). This rhu GMCSF is an altered form of the native molecule: the position 23 arginine has been replaced with a leucine to facilitate expression of the protein in yeast (Saccharomyces cerevisiae) and the carbohydrate moiety may be different from the native protein. The Clinical Center Pharmacy will purchase commercially available sargramostim. Rhu GM-CSF will be used both as an ancillary product for the maturation of dendritic cells and for administration to patients with peptide vaccination. 13.2.2 Formulation and Preparation: Lyophilized LEUKINE is a sterile, white, preservative-free powder (250 mcg) that requires reconstitution with 1 mL Sterile Water for Injection, USP or 1 mL Bacteriostatic Water for Injection, USP. Reconstituted lyophilized LEUKINE is a clear, colorless liquid suitable for subcutaneous injection or intravenous infusion. The vial of lyophilized LEUKINE contains 250 mcg (1.4 x 106IU/vial) sargramostim, 40 mg/mL mannitol, USP; 10 mg/mL sucrose, NF; and 1.2 mg/mL tromethamine, USP, as excipients. Biological potency is expressed in International Units (IU) as tested against the WHO First International Reference Standard. The specific activity of LEUKINE is approximately 5.6 x 106IU/mg.
figure 12 ad5f35 human her2/neu
13.2.3 stability and storage

Lyophilized LEUKINE should be refrigerated at 2 to 8 C (36 to 46 F) and is stable for at least eighteen months. It should not be frozen, shaken or used beyond the expiration date printed on the vial. LEUKINE reconstituted with 1.0 mL of Sterile Water for Injection, USP (without preservative) should be administered as soon as possible and within 6 hours following reconstitution. Reconstituted solutions prepared with Bacteriostatic Water for Injection USP
(0.9% benzyl alcohol) may be stored for up to 20 days at 2 to 8 C prior to use. The contents of vials reconstituted with different diluents should not be mixed together.
figure 12 ad5f35 human her2/neu
13.2.4 administration procedures

Lyophilized LEUKINE will be used *in vitro* by the Department of Transfusion Medicine for the generation of dendritic cells by the Department of Transfusion Medicine.  
13.2.5 Incompatibilities:
Refer to the package insert/PDR/Formulary.
figure 12 ad5f35 human her2/neu
13.3 interleukin-4 cellgenix 13.3.1 source

Interleukin-4 (IL-4) used in this study is investigational. It is manufactured and supplied by CellGenix (Master File cross reference BB-MF 11269). It will be used as an ancillary product to mature dendritic cells *in vitro* and will not be administered directly to patients. IL-4 exerts important effects on B cells, T cells, macrophages, eosinophils, hematopoietic progenitor cells, endothelial cells and promotes the maturation of dendritic cells. The complementary DNA clone (cDNA), when expressed in E.coli yields a 130 amino acid residules with a molecular mass of 14 kDa. IL-4 is a highly purified ( 97% as determined by SDS-PAGE), sterile, water-soluble protein. 13.3.2 Formulation and Preparation: RhIL-4 Sterile Powder for Injection is supplied in a 50mcg vial as a sterile lyophilized powder formulated with glycine, human serum albumin, citric acid, and sodium citrate. Approximately 1.2 mL of Sterile Water for Injection USP should be added to each vial of rhIL-4 Sterile Powder for Injection. The vial should be gently agitated to completely dissolve the powder and should be inspected visually for discoloration and particulates prior to use.
figure 12 ad5f35 human her2/neu
13.3.3 stability and storage

The product both intact and reconstituted vial should be stored at -20C to -80C and should be used within 24 hours. 13.3.4 Administration Procedures: To be used in dendritic cell culture, not administered directly to patients. 13.3.5 Incompatibilities: None known in culture.
figure 12 ad5f35 human her2/neu
13.4 klh (keyhole limpet hemocyanin)

KLH protein (Product Code KLH-02NV) was purchased from Stellar Biotechnologies, Inc. and then vialed by Element SL Pharma. It is stored in Center for Cellular Engineering in Department of Transfusion Medicine, NIH Clinical Center for use in dendritic cell culture. The KLH formulation to be utilized by NIH has been purified as native molecules and designated as High Molecular Weight KLH (HMW-KLH). The only source for KLH is the hemolymph of Giant Keyhole Limpets, Megathura crenulata, an ocean mollusk. Stellar's clinical grade HWM-KLH is purified under cGMP conditions, and sourced from giant keyhole limpets cultured by Stellar under controlled aquaculture systems. KLH protein is expressed in two subunit isoforms (KLH1 and KLH2) of approximately 360,000 and 400,000 monomeric molecular weight, respectively. The KLH monomers are each composed of 7 or 8 functional unit domains; each functional unit contains an oxygen binding site containing two copper atoms. Both KLH isoforms assemble into native homo-decamers and didecamers of 4,000,000 to 8,000,000 molecular weight in hemolymph. 13.4.2 Formulation and Preparation: Stellar high molecular weight KLH is provided in soluble form in a buffer solution that is composed of 10mM sodium phosphate, 135mM NaCl, 1mM CaCl2 and 0.5mM MgCl2 as a bulk drug substance at 5mg/mL and then diluted and vialed into single use vials at 2mg/mL, 250 µL/vial by Element SL Pharma, a vendor qualified by NIH Sterile Product for Human Administration (SPHA) committee. 13.4.3 Stability and Storage:
HMW-KLH is stable for at least 12 months when stored at 2 to 8C. A stability testing program is in place. 13.4.4 Administration Procedures: HMW-KLH will be used in vitro by the Department of Transfusion Medicine at a concentration of 10mcg/mL for the generation of dendritic cells. Any excess culture media will be removed Cells before administration. 13.4.5 Incompatibilities: No information available.
figure 12 ad5f35 human her2/neu
14 references

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12. Piccart-Gebhart MJ, Procter M, Leyland-Jones B, et al. Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. *New Engl J Med.* 2005;353(16):16591672.
13. Tan-Chiu E, Yothers G, Romond E, et al. Assessment of cardiac dysfunction in a randomized trial comparing doxorubicin and cyclophosphamide followed by paclitaxel, with or without trastuzumab as adjuvant therapy in node-positive, human epidermal growth factor receptor 2-overexpressing breast cancer: NSABP B-31. *J Clin Oncol.* 2005;23(31):7811-7819.  
14. Geyer CE, Forster J, Lindquist D, et al. Lapatinib plus capecitabine for HER2-positive advanced breast cancer. *New Engl J Med.* 2006;355(26):2733-2743.  
15. Koninki K, Barok M, Tanner M, et al. Multiple molecular mechanisms underlying trastuzumab and lapatinib resistance in JIMT-1 breast cancer cells. *Cancer Lett.* 2010;294(2):211-219.  
16. Baselga J, Cortes J, Kim SB, et al. Pertuzumab plus Trastuzumab plus Docetaxel for Metastatic Breast Cancer. *New Engl J Med.* 2012;366(2):109-119.  
17. LoRusso PM, Weiss D, Guardino E, Girish S, Sliwkowski MX. Trastuzumab Emtansine:
A Unique Antibody-Drug Conjugate in Development for Human Epidermal Growth Factor Receptor 2-Positive Cancer. *Clin Cancer Res.* 2011;17(20):6437-6447.  
18. Park JM, Terabe M, Steel JC, et al. Therapy of advanced established murine breast cancer with a recombinant adenoviral ErbB-2/neu vaccine. *Cancer Res.* 2008;68(6):1979-1987.  
19. Park JM, Terabe M, Sakai Y, et al. Early Role of CD4+ Th1 cells and antibodies in HER2 adenovirus-vaccine protection against autochthonous mammary carcinomas. J
Immunol. 2005;174:4228-4236.  
20. Rolla S, Nicolo C, Malinarich S, et al. Distinct and non-overlapping T cell receptor repertoires expanded by DNA vaccination in wild-type and HER-2 transgenic BALB/c mice. *J Immunol.* 2006;177(11):7626-7633.  
21. Cavallo F, Calogero RA, Forni G. Are oncoantigens suitable targets for anti-tumour therapy? *Nat Rev Cancer.* 2007;7(9):707-713.  
22. Sakai Y, Morrison BJ, Burke JD, et al. Vaccination by Genetically Modified Dendritic Cells Expressing a Truncated neu Oncogene Prevents Development of Breast Cancer in Transgenic Mice. *Cancer Res.* 2004;64:8022-8028.
23. Disis ML, Calenoff E, McLaughlin G, et al. Existent T-cell and antibody immunity to HER-2/neu protein in patients with breast cancer. *Cancer Res.* 1994;54:16-20.  
24. Disis ML, Knutson KL, Schiffman K, Rinn K, McNeel DG. Pre-existent immunity to the HER-2/neu oncogenic protein in patients with HER-2/neu overexpressing breast and ovarian cancer. *Breast Cancer Res Treat.* 2000;62(3):245-252.  
25. Finn OJ. Cancer immunology. *N Engl J Med.* 2008;358(25):2704-2715. 26. Kiessling A, Fussel S, Wehner R, et al. Advances in specific immunotherapy for prostate cancer. *Eur Urol.* 2008;53(4):694-708.  
27. Tanaka Y, Koido S, Chen DS, Gendler SJ, Kufe D, Gong JL. Vaccination with allogeneic dendritic induces antitumor immunity in cells fused to carcinoma cells MUC1 transgenic mice. *Clin Immunol.* 2001;101(2):192-200.  
28. Higano CS, Small EJ, Schellhammer P, et al. Sipuleucel-T. *Nat Rev Drug Discov.*
2010;9(7):513-514.  
29. Kantoff PW, Higano CS, Shore ND, et al. Sipuleucel-T immunotherapy for castrationresistant prostate cancer. *N Engl J Med.* 2010;363(5):411-422.  
30. Higano CS, Schellhammer PF, Small EJ, et al. Integrated Data From 2 Randomized, Double-Blind, Placebo-Controlled, Phase 3 Trials of Active Cellular Immunotherapy With Sipuleucel-T in Advanced Prostate Cancer. *Cancer.* 2009;115(16):3670-3679.
31. Ladjemi MZ, Jacot W, Chardes T, Pelegrin A, Navarro-Teulon I. Anti-HER2 vaccines:
new prospects for breast cancer therapy. *Cancer Immunol Immun.* 2010;59(9):1295-1312.  
32. Norell H, Poschke I, Charo J, et al. Vaccination with a plasmid DNA encoding HER2/neu together with low doses of GM-CSF and IL-2 in patients with metastatic breast carcinoma: a pilot clinical trial. *J Transl Med.* 2010;8.  
33. Mitsuhashi M. Ex vivo simulation of leukocyte function: Stimulation of specific subset of leukocytes in whole blood followed by the measurement of function-associated mRNAs. J Immunol Methods. 2010;363(1):95-100.  
34. Marshall DT, Savage SJ, Garrett-Mayer E, et al. Vitamin D3 supplementation at 4000 international units per day for one year results in a decrease of positive cores at repeat biopsy in subjects with low-risk prostate cancer under active surveillance. *J Clin* Endocrinol Metab. 2012;97(7):2315-2324.  
35. Wagner D, Nonn L, Antonio A, et al. Double-blind randomized clinical trial of vitamin D3 showing effects on tissue calcitriol levels, gene expression and proliferation immunohistochemistry in prostate cancer. *Cancer Res.* 2012;72.  
36. Peppone LJ, Rickles AS, Janelsins MC, Insalaco MR, Skinner KA. The Association Between Breast Cancer Prognostic Indicators and Serum 25-OH Vitamin D Levels. *Ann* Surg Oncol. 2012;19(8):2590-2599.  
37. Nishino M, Giobbie-Hurder A, Gargano M, Suda M, Ramaiya NH, Hodi FS. Developing a Common Language for Tumor Response to Immunotherapy: Immune-Related Response Criteria Using Unidimensional Measurements. *Clin Cancer Res.*
2013;19(14):3936-3943.  
38. Wolchok JD, Hoos A, O'Day S, et al. Guidelines for the Evaluation of Immune Therapy Activity in Solid Tumors: Immune-Related Response Criteria. *Clin Cancer Res.* 2009;15(23):7412-7420.  
39. Hodi FS, Hwu WJ, Kefford R, et al. Evaluation of Immune-Related Response Criteria and RECIST v1.1 in Patients With Advanced Melanoma Treated With Pembrolizumab. J Clin Oncol. 2016;34(13):1510-+.
figure 12 ad5f35 human her2/neu
15 appendices 15.1 appendix 1

| ECOG Performance Status Scale Grade Descriptions Normal activity. Fully active,  0 able to carry on all pre-disease  performance without restriction. Symptoms, but ambulatory.  Restricted in physically strenuous  activity, but ambulatory and able  to carry out work of a light or  sedentary nature (e.g., light  housework, office work). 1 In bed <50% of the time.  Ambulatory and capable of all  self-care, but unable to carry out  any work activities. Up and  about more than 50% of waking  hours. 2 In bed >50% of the time.  Capable of only limited self-care,  confined to bed or chair more  than 50% of waking hours. 3 100% bedridden. Completely  disabled. Cannot carry on any  self-care. Totally confined to bed  or chair. 4 5 Dead.   |
|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|  
15.2 APPENDIX 2: NEW YORK HEART ASSOCIATION(NYHA) FUNCTIONAL C**LASSIFICATION**
The New York Heart Association (NYHA) Functional Classification in a Patient with Heart Disease Overview: The New York Heart Association (NYHA) developed a functional classification for patients with heart disease.  
Patients: Heart disease must be present.  
Parameters:
(1) Limitations on physical activity
(2) Symptoms (undue fatigue palpitations dyspnea and/or anginal pain) with ordinary physical activity
(3) Status at rest  
| Limitations on              | Symptoms with Ordinary    | Status at Rest      | Class   |
|-----------------------------|---------------------------|---------------------|---------|
| Physical Activity           | Physical Activity         |                     |         |
| None                        | None                      | Comfortable         | I       |
| Slight                      | Symptomatic with ordinary | Comfortable         | II      |
| activities                  |                           |                     |         |
| Marked                      | Symptomatic at less than  | Comfortable         | III     |
| ordinary levels of activity |                           |                     |         |
| Unable to perform any       | Discomfort with any       | Symptomatic at rest | IV      |
| activity                    | activity                  |                     |         |  
In addition the level of objective evidence was also classified:  
| Objective Evidence of Cardiovascular Disease   | Class   |
|------------------------------------------------|---------|
| No evidence of disease                         | A       |
| Minimal disease                                | B       |
| Moderate disease                               | C       |
| Severe disease                                 | D       |
figure 12 ad5f35 human her2/neu
15.3 appendix 3 standard operating

The autologous AdHER2 DC vaccine will be manufactured by the Department of Transfusion Medicine (DTM) at the NIH Clinical Center in Bethesda, MD:
National Institutes of Health NIH Clinical Center Department of Transfusion Medicine 10 Center Drive-MSC 1184 Building 10, Room 1C711 Bethesda, MD 20892-1184 The Sponsor-Investigator held IND that will be submitted for this product will include a Letter of Authorization to Cross-Reference BB-MF 11054 from the Master File Type V for Facility, Operational and Quality Systems for the Manufacture of Cellular Therapy Products that is held by DTM.
figure 12 ad5f35 human her2/neu
autologous cell harvest

Blood collection will be by standard apheresis: up to 18L of whole blood will be processed in order to collect peripheral blood mononuclear cells (MNC) with a target yield of at least 2.2 ×109 monocytes. Lymphocytes will also be cryopreserved for immunologic assays. Apheresis will be performed in the Clinical Center (CC) Department of Transfusion Medicine (DTM) using approved standard operating procedures. Bilateral peripheral venous access or a venous dual lumen catheter will be used for apheresis. Prophylactic intravenous CaCl2 and MgSO4 infusions may be administered during apheresis to treat or prevent citrate toxicity at the discretion of the DTM physician per routine. Autologous monocytes for dendritic cell culture will be enriched from peripheral blood MNC apheresis collections by counter-flow elutriation, aliquoted into multiple vials and cryopreserved for future preparation of the dendritic cell products: target of at least 6 vials with ~ 333 x 106cells/vial. Additional apheresis will be offered per investigator's discretion if the collected cells or plasma are not sufficient to manufacture at least 5 vaccine doses. All cell processing will be conducted in accordance with approved DTM policies and procedures.
figure 12 ad5f35 human her2/neu
adher2 transduced dendritic cells

Autologous AdHER2 DC vaccines will be prepared under GMP conditions from cryopreserved patient monocytes as outlined in the accompanying AdHER2 DC manufacturing schema. The strength (vp/mL) of AdHER2 vector will be adjusted per most recent stability test result. The label on the vial reflects the strength at initial release. The range of  50% of the strength reported on the product label will be deemed acceptable to use in vaccine manufacturing with PI review of updated annual stability test result and approval.
figure 12 ad5f35 human her2/neu
15.4 appendix 4 adher2 dc

Description Of Adher2 **Dc Vaccine Manufacture**  
Day 0 Prior to the scheduled administration of the Dendritic Cells vaccine, one cryopreserved monocytes enriched aliquot will be thawed, washed and evaluated for cell count and viability. Monocytes will be resuspended in media containing 90% RPMI-1640, 10% auto HF plasma or allo HIF AB plasma, 2000 IU/mL IL-4 2000 IU/mL rGM-CSF and 10 mcg/mL gentamicin.  
Day 1 No processing required Day 2 Fresh media and cytokines (2000 IU/mL rIL-4, 2000 IU/mL of rGM-CSF) will be added to the culture; KLH will be added at a final concentration 10 mcg/mL.  
Day 3 The AdHER2 vector will be added at a ratio of 3000 viral particles per monocytes plated on Day 0; vector viral particles will be added at -24 hrs from harvesting.  
Vector transfection will occur over 4 hours before the addition of maturation cocktail at -20 hrs from harvesting. Maturation cocktail consist of LPS and IFN- at a final concentration of 30 ng/mL and 1000 IU/mL, respectively.
Removal of sample for -24 hours sterility testing.  
Day 4 Final harvest, packaging and lot release testing of autologous AdHER2 DC vaccine preparation. The final transfected, volume-reduced mature AdHER2 DC vaccine product will be prepared in sterile syringes for fresh administration intradermally.  
The AdHER2 DC vaccine product will be prepared as summarized above using a comprehensive, validated manufacturing process described in the DTM Standard Operating Procedures (SOP). Detailed SOP for processing, labeling, storage, and quality assays are also available on site in the Cell Processing Section of DTM. All DTM SOPs related to the manufacture of the AdHER2 DC vaccine will be included in the Sponsor-Investigator held IND to be submitted for the product.
figure 12 ad5f35 human her2/neu
vaccine processing steps and in

Multiple sample and assay timelines for the manufacturing processes of the AdHER2 DC vaccine are detailed in the CMC file in DTM and include:  
➢ Timeline for processing and in process testing of autologous mononuclear cells to generate monocyte and lymphocyte fractions via elutriation for cryopreservation.  
➢ Timeline for processing and in process testing during AdHER2 DC vaccine manufacture.
figure 12 ad5f35 human her2/neu
vaccine manufacture critical reagents

Critical reagents in the manufacture of autologous AdHER2 DC vaccine include interleukin-4 (IL-4), KLH (Keyhole Limpet Hemocyanin), sagramostim (GM-CSF) and reference endotoxin
(E. coli). Detailed information on these agents is outlined in protocol Section 13, Pharmaceutical Information. Letters of Authorization to cross-reference the master files of these agents will be included with the formal IND submission for the product.
figure 12 ad5f35 human her2/neu
vaccine release testing

Release testing will include not only safety testing (microbiological testing) but will also include any product characterization that is being done such as identity, purity, and potency testing. The following tests will be performed related to AdHER2 DC vaccine release criteria and quality control:
figure 12 ad5f35 human her2/neu
a) vaccine product safety testing

1. Sterility (bacterial and fungal) testing of all in-process and final product samples of the AdHER2 DC vaccine product will be done by the Bactec automated blood culture system. Although final product sterility testing will be performed, results will not be available as a release criterion. DTM has completed and published the results of a study demonstrating that both the BacT/Alert (BioMerieux) and the Bactec (Becton Dickinson) automated culture systems are superior to the CFR/USP method for sterility testing of cell therapy products (seeded study; *Khuu HM et al. Cytotherapy 2004; 6(3): 183-195*). They also published a parallel testing study of CFR/USP vs. Bactec, for sterility testing of actual cell therapy products from their facility. Results of the parallel studies show that the automated culture methods had fewer false positives than CFR/USP (*Khuu HM et al. Transfusion 2006; 46:* 2071-82). Based on the analysis of both studies, DTM has sufficient data to implement the Bactec system as the replacement for the CFR/USP method. Justification for use of automated system in place of CFR/USP method is based on the results of the studies described above. In the seeded study, 10 different organisms were seeded into mononuclear cell products suspended in 6 different media, one of which was X-Vivo20 (which contains gentamicin), and one of which was our standard freeze mix (Plasmalyte A, human serum albumin, Pentastarch, DMSO, Dornase, and heparin), which is used to cryopreserve all products in this protocol. The antibiotic level tested in the seeded study is **50mg/mL** and this level was shown to be non-inhibitory in the automated culture system. This is the same level in the samples for sterility for the product in this IND.  
2. Negative gram stain must be documented 3. Sterility of -24hrs in process sample must be documented 4. Mycoplasma testing will be done by Mycoplasma PCR method (Roche) but results will not be available for use as a release criterion 5. Endotoxin testing will be done by the KQCL method; endotoxin level must be < 5EU/ml. 6. Transfusion-transmitted disease (TTD) testing will be performed using methods described in our Master File CC-MF-11054 (Attachment 8-A), with the following modification. Testing is performed by Blood Systems Laboratories in Tempe, AZ CLIA Reg \#03D0911463). Agents include T. cruzi as well as those listed. Syphilis is tested using PK-TP assay, instead of RPR.
figure 12 ad5f35 human her2/neu
b) vaccine product characterization

1. Total cell number as determined using an automatic counter. 2. Viability using Trypan blue dye exclusion with a minimum viability of > 60% as release criteria.  
3. Cell phenotype and expression for HER2neu by FACS.  
Phenotype markers will include: **CD14, CD38**, CD54, CD83, CD86, **CD 340** (HER2neu), CCR7, HLA-DR. **Key phenotype markers in bold.**
4. Day 3 in process DC supernatants prior to transfection with adenovirus for cytokine assessments by multiplex and proteomic array (for research purposes).
figure 12 ad5f35 human her2/neu
vaccine product release and action

General action plans for dealing with positive tests for sterility, mycoplasma, and endotoxin testing are presented in our Master File BB-MF-11054 (Attachments 9-B through 9-D). As noted above, not all safety testing will be available prior to release of the AdHER2 DC vaccine product. For testing with final results available, results must be within pre-specified limits and documented on the Certificate of Analysis before the product is released for administration to the patient.
figure 12 ad5f35 human her2/neu
vaccine final product preparation steps

The final AdHER2 DC vaccine product will be harvested from the 5-day culture product and packaged for fresh infusion on the same day.
figure 12 ad5f35 human her2/neu
vaccine product tracking and labeling

The system for labeling and tracking of all products in this IND will be the same as the general system used in the DTM Cell Processing Facility, described in their Master File BB-MF-11054, section XI.
figure 12 ad5f35 human her2/neu
vaccine product administration

Autologous AdHER2 DC vaccines will be harvested from the 5-day culture product as previously described and packaged for fresh administration on the same day according to DTM
Standard Operating Procedures. The protocol specified AdHER2 DC vaccine dose of 5 X 106 viable cells, 10 X 106*viable* cells, 20 X 106*viable* cells or 40 X 106*viable* cells will be concentrated in a final total volume of 1.0mL for intradermal administration immediately upon receipt in the clinical setting. Post packaging testing of DC vaccines produced by DTM indicates that the product should be stable for at least 2 hours. Patients will have vital signs monitored for one hour following administration of the first dose of AdHER2 DC vaccine and for 15 minutes with all subsequent doses if there are no significant acute adverse systemic clinical events (e.g. fever, hypotension, wheezing, rash/urticaria) documented after the first dose. It is anticipated that the primary adverse events associated with AdHER2 DC vaccination will be local injection site reactions, and possibly symptoms related to local anti-tumor inflammatory responses at sites of primary and metastatic disease including focal organ discomfort and/or reactive pleural and peritoneal effusions. 15.5 APPENDIX 5: ADHER2 DENTRITIC CELL VACCINE REPORT **CARD** Date/Time Vaccine Administered: ____________ Administered By: _________________
Study Part I Part II Vaccine Dose 10e6 cells 20e6 cells 40e6 cells Study Week __________ Vaccine Dose \# 1 2 3 4 5  
| Injection  Site:  Forearm                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   | Injection  Volume (mL)   | ID  Wheal  Present?   | DA              | DA              | DA              | DA              | DA              | Date   |
|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------|-----------------------|-----------------|-----------------|-----------------|-----------------|-----------------|--------|
| Y 1                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         | Y 2                      | Y 3                   | Y 4             | Y 5             | resolved        |                 |                 |        |
| Left                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       | Present                 |  0                   |                 |                 |                 |                 |                 |        |
| Site #1                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     | Right                   | Absent               |  1  2  3     |  0  1  2  3 |  0  1  2  3 |  0  1  2  3 |  0  1  2  3 |        |
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| Total Injection Volume: _______________    Injection Site Reaction Grading (NCI CTCAE v4.03)  0 No Reaction  1 Tenderness with or without associated warmth, redness, itching.  2 Pain, swelling (edema), lipodystrophy (shrunken area of the skin due  to loss of fat), or phlebitis (inflammation of veins in the skin).   3 Ulceration or necrosis (skin break down); severe tissue damage.  Operative intervention indicated.  4 Life-threatening consequences. Urgent intervention indicated.Note: If you have a reaction that lasts longer than 5 days please continue to describe it on  the back of this page. |                          |                       |                 |                 |                 |                 |                 |        |  
Print Name: Signature:
figure 12 ad5f35 human her2/neu
15.6 appendix 6anti-her2 antibody and

The majority of the testing on this study will be done in the NIH Clinical Center clinical laboratory following their guidelines for blood collection and tube type. The appropriate tube for uncommon laboratory tests and immunologic research specimens and where they should be sent are as follows: Quantitative Anti-HER2 Antibody Testing: Weeks 0, 8, 16, 24, 32, 40, 48, 60, 76, 100, and
figure 12 ad5f35 human her2/neu
124

Purpose: To determine the immunogenicity of autologous Ad HER2 dendritic cell vaccination as measured by a 3-fold increase in anti-HER2/neu antibody concentration (measured as mcg/mL) or a 4-fold increase in antibody dilution titers over baseline.
Specimen Processing: 1 10mL Red Top Clot activator Specimens will be processed by the Clinical Support Laboratory Leidos Biomedical Research, Inc. Serum will be aliquoted into vials and cryopreserved until ready for interrogation in batched specimen assays.
figure 12 ad5f35 human her2/neu
characterization of vaccine-induced antibody profiles

Purpose: To characterize vaccine-induced antibody profiles using HER2 peptide microarrays, examining reactivity to HER2 extracellular (EC), transmembrane (TM) and intracellular (IC) domains to document evidence of epitope spreading. To characterize immunoglobulin sub-classes of anti-HER2 antibodies.
Specimen Processing: No additional specimens to be drawn.  
Will utilize clinical specimens drawn and processed for quantitative anti-HER2 antibody testing.
figure 12 ad5f35 human her2/neu
cellular responses to her2/neu weeks

Send via Frederick Courier to NCI Frederick Clinical Support Laboratory for specimen processing and freezing.  
Tetramer Assay: Will be performed by the flow cytometry unit in the NCI Frederick Clinical Support Laboratory or Vaccine Branch Flowcytometry Core.  
IFN- **ELISPOT Assay:** Will be performed in the Laboratory of Cell Mediated Immunity or Vaccine Branch Flowcytometry Core.
The Clinical Support Laboratory, Leidos Biomedical Research, Inc., processes and cryopreserves samples in support of IRB-approved, NCI clinical trials. The laboratory is located in a controlled access building and laboratory doors are kept locked at all times. Upon specimen receipt, each sample is assigned a unique, sequential laboratory accession ID number. All products generated by the laboratory that will be stored either in the laboratory freezers or at a central repository facility are identified by this accession ID. An electronic database is used to store information related to patient samples processed by the laboratory. Vial labels do not contain any personal identifier information. Samples are stored inventoried in locked laboratory freezers and are routinely transferred to the NCI-Frederick repository facilities for long-term storage. These facilities are operated by ATCC. under subcontract to Leidos Biomedical Research, Inc. Access to stored clinical samples is restricted. Investigators establish sample collections under "Source Codes" and the investigator is responsible for the collections, typically the protocol Principal Investigator who has access to the collection. Blood and tissue specimens collected in the course of this research project may be banked and used in the future to investigate new scientific questions related to this study. However, this research may only be done if the risks of the new questions were covered in the consent document, the patient has provided consent for the research use of their clinical specimens, and the proposed research has undergone prospective IRB review and approval. The NIH Intramural IRB will be notified when samples are destroyed. 15.7 APPENDIX 7EVALUATION OF ANTI-AD5 ANTIBODY I**MMUNITY** An intramural scientific collaborator or scientific contractor to perform these studies to be determined. Anti-Ad5 antibody ELISA assessments will be performed at Weeks 0, 8, 16, 24, 32, 40, 48, 60, 76, 100, and 124 to determine **in an exploratory manner if:**  
1. AdHER2 DC vaccination is associated with a measurable increase in anti-Ad5 antibody over time.  
2. Pre-existing levels of anti-Ad5 antibodies inversely correlate with vaccine immunogenicity as measured by anti-HER2 antibody levels.
We will store the samples in a -80°C freezer until batched shipped at which point they will be processed.
figure 12 ad5f35 human her2/neu
15.8 appendix 8characterization of soluble

Testing and characterization of soluble HER2 and anti-HER2 antibodies will be performed by a third party Midissia or other third-party collaborator using samples stored as described in Appendix 6. Midissia is a developing biopharma company that has licensed the AdHER2 DC
platform in combination with a checkpoint inhibitor.  
15.9 APPENDIX 9EXPLORATORY CORRELATIVE S**TUDIES** 1 of 2 Measurement of Function-Associated mRNAs in Whole Blood: Hitachi Chemical Function-Associated mRNAs: One 6 mL green top sodium heparin tube
(Week -0)
figure 12 ad5f35 human her2/neu
for prediction of responses

➢ Blood will be drawn into one 6 mL green top tube.  
➢ Add 180 L each of blood into one 8-well strip. Strips contain PHA, HAG, PBS, DMSO,
rIL2, CEF, LPS/ZA, and HER2).  
➢ Incubate at 37C for 4 hours.  
➢ Store in -80C (+/- 10C) freezer until ready for shipment to Hitachi Chemical
➢ Note: specimen should be drawn DAY OF VACCINE *before 1pm* to allow adequate time for specimen processing and four hour incubation by Trepel lab staff.  
For Characterization of Responses will be assessed after "Prediction of Responses" described
figure 12 ad5f35 human her2/neu
above confirms the list of

➢ Blood will be drawn into one 6 mL green top tube.  
➢ Approximately 1% (2x105cells) of final AdHER2 DC vaccine product in 0.1mL of standard suspension (taken at the same time as DC product QC test samples) and 0.2mL of heparinized whole blood will be placed into conical freezer microtubes in the following manner:
1. **Tube 1:** 0.1mL of 2x105cells of final AdHER2 DC vaccine product *only* 2. **Tube 2**: 0.1mL of 2x105cells of final AdHER2 DC vaccine product *plus* 0.2mL
of heparinized whole blood.  
3. **Tube 3**: 0.2mL of heparinized whole blood *only*
➢ Incubate at 37C for 4 hours.  
➢ Store in -80C (+/- 10C) freezer until ready for shipment to Hitachi Chemical ➢ Note: specimen should be drawn DAY OF VACCINE *before 1pm* to allow adequate time for specimen processing and four hours incubation by Trepel lab staff.  
2 of 2 Multiparameter Flow Cytometric Analysis of Circulating Tumor Cells (CTC) and Immune Cell Subsets: Jane Trepel, DTB Analysis will be performed by the laboratory of Jane Trepel. CTC cells and immune cell subsets will be identified by mulitparameter flow cytometry. The order of priority for immune subset analysis is T, B, NK, NKT, Tregs, MDSC and dendritic cells.  
CTC Analysis: One 10mL lavender top tube
(Weeks 0, 8, 16, 24, 32, 40, 48, 60, 76, 100, and 124 when patient schedule permits collection before 1pm.)
Immune Cell Subset Analysis: One 10mL lavender top tubes
(Weeks 0, 8, 16, 24, 32, 40, 48, 60, 76, 100, and 124 when patient schedule permits collection before 1pm.) Multiparameteric flow cytometry using a Miltenyi Quant flow cytometer. The order of priority for immune subset analysis is T, B, NI, NKT, Tregs, MDSC and dendritic cells.  
NKT Cell Analysis: One 10mL lavender top tubes
(Weeks 0, 12 (s/p 3 doses of vaccine), 28 and 48) Sample Logistics *for both Function-Associated mRNAs and CTC and Immune Subsets:*
➢ **Notify the Trepel lab via email** when the clinical sample is scheduled to be drawn:
o Sunmin Lee (lees@pop.nci.nih.gov) o Min-Jung Lee (leemj@mail.nih.gov)
o Jane Trepel (trepel@helix.nih.gov)
➢ Label the clinical specimen tubes and *include the study week number*.  
➢ Note: *specimen should be drawn before 1pm* to allow adequate time for four hours incubation for the Hitachi clinical specimens.  
➢ **Phone the Trepel lab at 240-760-6330** when the specimen is drawn for pick up by the Trepel lab.  
➢ The laboratory of Jane Trepel where specimens will be processing and cryopreserved is in Bldg.10, Rm. 12C208
figure 12 ad5f35 human her2/neu
immune and molecular profiling through

Briefly, patient blood samples collected at baseline and at each time point when vaccine is administered is analyzed via next generation sequencing and RNA sequencing with the aim of identifying potential biologic/immunological correlates of vaccine mediated efficacy, including but not limited to the potential upregulation of checkpoints using advanced bioinformatics tools. Subcontractor: Novogene Corporation, Inc. 8801 Folsom BLVD, Suite 290, Sacramento, CA 95826 by Midissia Therapeutics, Inc. Specimen Processing:. 10ml of whole blood will be collected in should be in 2 Tempus RNA tubes at Weeks 0, 8, 16, 24, 30, and 48 and kept frozen at -80°C in Frederick Leidos lab until analysis. If Tempus tube sample is not available, any unused PBMNC frozen aliquots for cellular response evaluation can be used as an alternative. RNAseq will be performed on patient blood samples to provide a comparative analysis of gene expression at baseline and following treatment. This will provide a longitudinal scan of patient response to treatment. To ensure Quality control, RNA
quantity and quality are assessed using OD260 and OD260/280 ratios. RNAseq read quality is also subject to diligence. Each patient sample will have the entire transcriptome read and comparisons to baseline will be made by number of reads at each gene. Significant changes will be determined by a threshold p-value and a list of Differentially Expressed Genes (DEG) will then be assembled. DEG gene lists will then be used as input for *post-hoc* analysis. *Differential Expression analysis* provides data on the number of genes whose expression changes over the course of treatment and is presented by Venn diagram or volcano plot. *Gene Set Enrichment Analysis* uses the lists of differentially expressed genes (DEG) and compares them to sets of curated gene sets in 4 databases.  
- Gene Ontogeny (GO): a curated gene set associated with normal cell processes, such as: regulation of cell *motility.*
- Disease Ontogeny (DO): a curated gene set associated with disease processes, such as:
thrombocytosis
- Kyoto Encyclopedia of Genes and Genomes (KEGG): a curated gene set associated with signal transduction pathways.  
- Reactome: Another database of signal transduction nodes.  
DEG lists from patients will be used to identify genes associated with signal transduction pathways, cellular processes and disease states to identify individual genes or groups of genes associated with a gene set of interest. Those genes whose expression patterns change predictably with clinical response will ultimately be used as a biologic/immunological correlate of vaccine mediated efficacy.
figure 12 ad5f35 human her2/neu
15.10 appendix 10ad5f35her2ectm (adher2) vector

GMP Production Outline
(1 of 2)  
  
Adenoviral Vector GMP Production Outline
(2 of 2)
15.11 appendix 11map of the
ad5f35her2ectm (adher2) detailed vector manufacturing

15.12 APPENDIX 12 ADHER2 DENDRITIC CELL VACCINE CANCER TREATMENT HISTORY  
| Brief Cancer History:  Cancer Diagnosis Date of Diagnosis Stage of Cancer   |
|-----------------------------------------------------------------------------|  
Surgical Procedures:  
Operative Procedure Date **Comments**  
Prior Chemotherapy:  
Agent/Regimen Date/Duration Comment/Reason for D/C  
Prior HER2-Targeted Therapies: (trastuzumab, lapatinib, pertuzumab) PATIENT IS NOT ELIGIBLE FOR THIS STUDY IF POSITIVE FOR HER2-RELATED CARDIAC TOXICITY Prior Radiation Therapy  
| Prior HER2-Targeted Therapies: (trastuzumab, lapatinib, pertuzumab) Agent Date/Duration   | Comment/Reason for D/C   |    |
|-------------------------------------------------------------------------------------------|--------------------------|----|
| HISTORY OF HER2 THERAPY RELATED CARDIAC DYSFUNCTION                                       | YES                      | NO |
| (If yes document name of agent and describe toxicity and intervention)                    |                          |    |  
| Version Date: 07/22/2019   |               |         |
|----------------------------|---------------|---------|
| Therapy Type               | Date/Duration | Comment |
15.11 appendix 11map of the
pathology her2/neu expression part i

| HER2 Detection Method        | Negative    | Equivocal          | Positive    |
|------------------------------|-------------|--------------------|-------------|
| Vysis FISH Ratio             | Ratio < 1.8 | Ratio 1. 8 - < 2.2 | Ratio > 2.2 |
| (Measures HER2 oncogene) IHC | 0           | 1+ - 2+            | 3+          |
| (Measures HER2 protein)      |             |                    |             |  
Part I: Adults ≥ 18 with recurrent or progressive, metastatic solid tumors characterized by some HER2/neu expression that have failed standard therapies with known benefit but for whom trastuzumab is not *clinically indicated.*
Part II: Breast cancer patients with 3+ HER2/neu expression by IHC or a positive or an equivocal or positive FISH result with prior exposure to trastuzumab, lapatinib or *pertuzumab.*
Additional Comments:  
| Part I or Part II    | HER2 Detection   | Expression   | Date of   | Received from NIH or   |
|----------------------|------------------|--------------|-----------|------------------------|
| Method               | Collection       | Outside Lab  |           |                        |
| Vysis FISH Ratio IHC |                  |              |           |                        |  
Person Completing This Form:
15.11 appendix 11map of the
15.13 appendix 13 patient recruiting

In accordance with standard IRB regulations requiring approval of patient recruitment materials, The following script is for an informational video that will be posted to the NCI
YouTube Network sponsored by the NCI Office of Communications at:
www.youtube.com/ncigov. The informational video will feature Dr. Lauren Wood, the former Principal Investigator of the AdHER2 DC vaccine study and will be approximately 2-4 minutes in duration. The video link will be embedded in all standard patient recruitment materials sent out by the NCI CCR Office of Patient Outreach and Recruitment as well as the NIH Clinical Center Office of Communications, Patient Recruitment and Public Liaison. This informational video is a novel patient recruitment tool that is being piloted with this study and will be assessed for its impact on rates of study accrual.
15.11 appendix 11map of the
adher2 dc vaccine informational video

Hello. My name is Dr. Lauren Wood and I am the Principal Investigator of a new therapeutic cancer vaccine study that is being conducted at the NIH Clinical Center in Bethesda, MD. Therapeutic cancer vaccines are given to patients that already have cancer and are designed to train the immune system to better recognize and kill cancer cells. The hope is that once trained, cells of the immune system may be able to seek out and destroy cancer cells in areas of the body where chemotherapy drugs and radiation may not. Therapeutic vaccines may also continue to have an anti-tumor effect that persists even after the vaccine is no longer being given. The clinical trial we are conducting is studying a cancer vaccine that is being given to humans for the first time. The science behind the trial is based on animal models that showed that a single dose of a mouse vaccine, similar to the human vaccine being used in this study, could cure large established tumors in mice.  
The cancer vaccine is designed to target a tumor protein called HER2. The HER2 protein makes tumors more aggressive and more likely to spread. As a result, patients with HER2 positive tumors often don't do as well. One of the most common tumors associated with HER2 expression is breast cancer, although only about 25 percent of women with breast cancer have HER2 positive tumors. In addition, there are other tumor types that express the HER2 protein, including bladder, colon, lung, kidney, ovarian, and prostate cancers. Because the HER2 protein makes tumors more aggressive, a number of different drugs targeting it have been developed to treat cancer. One of the first drugs was trastuzumab, also known by the brand name Herceptin. It is approved for the treatment of breast and stomach cancer. As a monoclonal antibody, trastuzumab recognizes a distinct, small portion of the HER2 protein. Although it's been shown to benefit patients through multiple studies, some patients do not respond to treatment, some patients become resistant to treatment and still others don't have high enough levels of the HER2 protein in their tumors to benefit from trastuzumab treatment. There are also other drugs that are licensed or in development that target HER2 by different mechanisms. The cancer vaccine in this trial uses cells from the patient's own immune system to generate a custom-made vaccine specific for each patient. The immune system cells we use are called dendritic cells. A "dead" virus, that is not able to make copies of itself, is used to deliver the HER2 genetic material inside the dendritic cells. This causes cells to express the HER2 protein on their surface and the dednritic cells are then given back to the patient as a vaccine. The hope is that this custom-made cancer vaccine will be able to stimulate the patient's own immune system to make their own antibodies to HER2, and that these antibodies will recognize multiple sites of the HER2 protein, rather than just one limited site like trastuzumab. The primary goal of this study is to determine whether or not the vaccine is safe in human beings and determine if there is an optimal dose to give. We also want to know whether or not the vaccine stimulates the immune system to produce antibodies and whether these antibodies will help slow the growth of or possibly even shrink tumors. A very small percentage of people who are treated with trastuzumab develop cardiac toxicity. We don't have a good understanding of what causes the changes in heart function and we don't know whether or not the HER2 vaccine we are testing will have any adverse effects on the heart. It is why if you choose to participate in this study, you will be monitored very closely during the study and for some time even after your last dose of vaccine. Importantly, there have been multiple clinical trials of HER2 cancer vaccines and to date, none of them have been associated with the development of problems in heart function. Initially we are seeking patients who have recurrent disease with tumors that have some level of HER2 expression but who have never been treated with trastuzumab or any other HER2-directed therapies and do not qualify to be treated with these therapies. Multiple tumor types are allowed. Once we have established safety of the vaccine in this population, we will evaluate the vaccine in patients with breast cancer that have progressed despite treatment with trastuzumab and other HER2 therapies. If you would like further information about this clinical trial, please call 888-XXX-YYYY to speak with a research referral coordinator. Again, my name is Dr. Lauren Wood of the NCI Vaccine Branch in Bethesda, MD. Thank you for taking the time to watch this video and for your interest in our clinical study.  
Current run time with normal voice cadence and pace: 5 minutes 8 seconds.  
Further script editing and reduction in duration may occur following review by NCI Communications. If successful, this Principal Investigator informational video link could be replicated for multiple other clinical trials being conducted by the NCI CCR, the Clinical Center and other NIH institutes. Contact Information **of Key Partners for Proposed Pilot Video Recruitment Tool:**
James Alexander Public Affairs Specialist Office of Partnerships and Dissemination Initiatives Office of Communications and Education National Cancer Institute, NIH 6116 Executive Blvd, oom 4117 Bethesda, MD 20892-8351 Phone: 301-402-4917 Email: alexandj@mail.nih.gov Dinora Dominguez Chief, Patient Recruitment and Public Liaison Section Office of Communications, Patient Recruitment and Public Liaison NIH Clinical Center National Institutes of Health Department of Health and Human Services 10 Cloister Ct., Bldg 61 Bethesda, MD 20892-4754 Direct Line: 301-402-6072 Fax: 301-480-8640 Toll-Free Patient Referral Line: 1-800-411-1222 TTY: 1-866-411-1010 Internet: http://www.cc.nih.gov Study Information: http://clinicalstudies.info.nih.gov Twitter: NIHClinicalCntr Facebook: www.facebook.com/NIHClinicalCenter YouTube: www.youtube.com/user/NIHClinicalCenter Audio Podcasts: clinicalcenter.nih.gov/podcast Susan McMullen, RN Office of the Clinical Director Center for Cancer Research, NCI 9030 Old Georgetown Road Building 82, Room 101 Bethesda, Maryland 20892-8200 Phone: 301-402-5931 FAX: 301-594-7951 mcmulles@mail.nih.gov Hoyoung Maeng, M.D. Vaccine Branch, NCI Bldg. 10, Rm. 3B37 10 Center Drive, MSC 1578 Bethesda, MD 20892-1578 Phone: 240-781-3253 FAX: 301-480-8514 Email: hoyoung.maeng@nih.gov 13-C-0016 INFORMATIONAL VIDEO WEBLINKS: Healthcare Provider Informaional Video (run time 3:21): https://youtu.be/TxXsflkDfYQ Patient Informational Video (run time 3:25): https://youtu.be/ejscyiHe5t0 These informational videos are also posted on the following web sites: Dr. Wood's CCR Bethesda Trials Investigator Profiles web site: http://ccr.cancer.gov/staff/staff.asp?profileid=5570 In the CCR News web site Multimedia section: http://ccr.cancer.gov/news/InTheirOwnWords/Default.aspx Additional informational materials provided to patients: 13-C-0016 Science in a Snapshot Slide Deck NOTE: These "snap shots" and their content are taken directly from the informational videos that were approved by the NCI IRB on 02/25/13.  
  
- The HER2 protein is over-expressed in breast and many other types of cancers.  
- It sends signals to tumor cells to make them grow and prevents them from dying. - Tumors that over-express HER2 are associated with more aggressive disease, higher recurrence rates and reduced survival rates.  
1  
  
- Trastuzumab (Herceptin®) and pertuzumab (PERJETA™) are monoclonal antibodies that each recognize, a distinct, small portion of the HER2 protein.  
- These monoclonal antibodies have to be given repeatedly to have an effect.  
Despite their effectiveness in some cancers, trastuzumab and pertuzumab don't
.  
work for everyone and even when they are effective,  they may stop working after some time.  
  
Additional materials for posting on social media sites in accordane with NIH Social Media Policy and coordinated by the NIH Clinical Center Patient Recruitment and Public Liaison Office (Dinora Dominguez) and the NCI Office of Communications and Education (James Alexander):
15.11 appendix 11map of the
13-c-0016 social media site language

NIH vaccine researcher is studying individuals with bladder, colon, lung, ovarian, prostate, and kidney cancer. Watch Dr. Lauren Wood's videos to learn how vaccines are involved in cancer research. More information Video for Patients: https://youtu.be/ejscyiHe5t0 Video for Healthcare Providers https://youtu.be/TxXsflkDfYQ
Homepage slide Phase 1 trial for breast, bladder, colon, lung, ovarian, prostate, kidney and other cancers that express HER2. Watch video to learn about vaccines in cancer research. More information Video for Patients:
https://youtu.be/ejscyiHe5t0
15.11 appendix 11map of the
twitter govdelivery

135 characters- NIH vaccine researcher studying breast, bladder, colon, lung, ovarian, and other cancers. Watch videos to learn how vaccines are involved in cancer research. OR 129 characters- Watch videos for a cancer vaccine clinical trial for patients with HER2+ solid tumors including breast, bladder, colon, lung, ovarian and other cancers.
15.11 appendix 11map of the
or

131 characters- NIH vaccine researcher is studying individuals with bladder, colon, lung, ovarian, prostate, and kidney cancer. Watch Dr. Lauren Wood's vaccine videos. (this is more plain language and patients will be able to quickly relate to one of the following cancers)
15.11 appendix 11map of the
facebook

Dr. Hoyoung Maeng, a NCI vaccine researcher, recently became the PI of a Phase I trial to evaluate the safety and immune response to a therapeutic cancer vaccine designed to stimulate the immune system to recognize the HER2/neu gene in patients with solid tumors including breast, bladder, colon, lung, ovarian, prostate and other cancers. Video for Patients: https://youtu.be/ejscyiHe5t0 Video for Healthcare Providers:
https://youtu.be/TxXsflkDfYQ