For specifics on drug administration see section 11.1.5 . Vital sign assessments (temperature, heart rate, blood pressure, respiratory rate) are to be obtained after the subject has been in a seated resting position for at least 5 minutes. For the first injection, vital signs must be assessed 30 and 60 minutes after the injection. Vital signs must be assessed 30 minutes after the subsequent injections . 3.3 DOSE DELAY OR DISCONTINUATION Applies to events occurring outside of the DLT evaluation period • Patients must have recovered to ≤ grade 2 for injection site reaction or grade ≤ 1 for any other toxicity related to the vaccine s for the parameters used to assess levels of organ function requi red for eligibility (see Section 2) after each vaccination in order to receive a s ubsequent vaccination. Patients will receive a diary card to record injections site reactions (see Appendix B). If ≥ grade 3 nonautoimmune toxicity attributable to the vaccine s persists for > 42 days, the patient will not receive further vaccine inoculations and will be removed from the protocol treatment and followed for resolution of toxicity and immune/survival endpoints. • Patients who develop any ≥ grade 3 autoimmunity, not related to a therapeutic response, will be removed from the protocol treatment and followed for resolution of toxicity and immune/survival endpoints. • Patients who develop any grade 4 toxicity attributable to the vaccine s will be removed f rom the protocol treatment and followed for resolution of toxicity and immune/survival endpoints. • Dosing of the vaccine s injections should be given on schedule every 3 weeks (Week 0, 3, and 6) , and then bi-monthly (every 8 weeks) boosts for up to a year. In the event of confl icts, a +/ - 1 week window is acceptable. • For unrelated acute illnesses present at the time of a scheduled vaccination, dosing can be delayed u ntil symptoms subside, or the subject may be withdrawn at the discretion of the Investigator; delays up to 4 weeks are considered acceptable in this setting. 3.4 DOSE MODIFICATIONS No dose modifications are allowed with th ese vaccine s except for dose de-escalation as described in sectio n 3.1. Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 34 3.5 STUDY CALENDAR Assessment Screening Treatment (Every 3 -Week Dosing) Bimonthly_ Boosters (Every 8 -Week Dosing) End of Treatment (within 90 days after the last vaccine ) m Follow Upl Study Week Day -28 to -1 n Baselin e/0 3 6 14 22 30 38 46 54 Clinic Visit X Xc X X X X X X X X X Informed Consent Xa Inclusion/Exclusion X Demographics X Confirmation of diagnosis (path report) X Physical Examination, Heightb, Weight, ECOG Xc Xc X X X X X X X X X Medical Historyd X Xc Concomitant Medications X Xc X X X X X X X X X Vital Signse X Xc X X X X X X X X X 12-Lead ECG X X X X X X X X Confirm Contraceptive Measuresf X Study Drug Injection/ In jection Site Reaction Monitoringg X X X X X X X X X X Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 35 Assessment Screening Treatment (Every 3 -Week Dosing) Bimonthly_ Boosters (Every 8 -Week Dosing) End of Treatment (within 90 days after the last vaccine ) m Follow Upl Study Week Day -28 to -1 n Baselin e/0 3 6 14 22 30 38 46 54 Dispensation of Subject Diary Cardh X Review of Sub ject Diary Cardh X Tumor Imagingi X X X X X X X X Pregnancy Testj Xc,n Xc X X X X X X X X X Urinalysis X n X Chemistry Panel Xc,n Xc X X X X X X X X X CBC, Differential, Platelets Xc,n Xc X X X X X X X X X Coagula tion X n X Serum Virology (HIV, HBV, HCV)k X n Adverse Events X X X X X X X X X X Exploratory Immune Analysis X X X X X Telephone Follow Up X a Informed consent may be obtained on D0 after eligibility is confirmed and other screening protocols may be used to obtain screening labs and procedures as per NCI’s internal SOP b Height will only be assessed at screening. c If the assessment is performed within 24 hours prior to the first dosing, a second assessment at baseline/week 0 can be omitted. Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 36 d Complete medical history will be evaluated at screening and includes current and past cardiac and pulmonary history, document ation of diagnosis including history of current and prior cancers, prior treatment(s), and prior radiologic studies. Any new events in the medical history will be evaluated at b aseline.. e Vital signs include temperature, heart rate, blood pressure, and respiratory rate. Vital sign assessments are to be obt ained after the subject has been in a seated resting position for at least 5 minutes. For the first injection, vital signs must be assessed 30 and 60 minutes after the in jection. Vital signs must be assessed 30 minutes after the subsequent injections. f Acceptable contraceptive measures are described in Section 2.1.1.12 . g Injection site reactions will be monitored h Subjects will be given a diary c ard for the self -evaluation and reporting of injection site reactions. i All baseline tumor measurements should be performed based on the subject’s qualifying scan obtained within 28 days prior to t he start of treatment. Tumor imaging and assessments will be performed j Serum pregnancy test will be performed on females of childbearing -potential and women < 12 months since the onset of menopause. The pregnancy test will be performed at each dosing visit with the result confirmed as negative prior to dosing. k Serum virology test for HIV (as determined by ELISA and confirmed by western blot), and HBV and HCV (as determined by HBsAg a nd hepatitis C serology). l After the subject completes or withdraws from study therapy , the study team will contact the subject approximately every 3 months for 12 months and then approximately every 6 months for 24 months and then every 12 months thereafter for another 24 months to collect follow -up information, including survival status and any current cancer treatment regimen. m If the patient cannot return to the Clinical Center within 90 days after the last vaccine, the patient will be contacted by phone to AEs and laboratory assessments will be deferred until a later point if at feasib le. n See section 2.2 for screening criteria which are exceptions to the D -28 to D -1 screening window period. Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 37 3.6 INJECTION SITE REACTIONS Local injection site reactions are expected to occur over several days after vaccination. Erythema and some soreness are possible. Injection s ite reactions will be monitored by clinic staff prior to discharge and by subject self-evaluation and re porting. Subjects will be given a d iary card with reporting fields for diameter of erythema and duration for the seven days after the first set of vaccine injections (see Appendix B). In addition, pain at the injection site, fever, and chills will be collected as Yes/ No and medications ( over-the-counter ibuprofen, acetaminophen , aspirin , etc.) taken for injection site p ain or discomfort as Yes/No. The site staff will review the diary cards with the subject during the next clinic visit and record responses in the case report form. 3.7 CRITERIA FOR REMOVAL FROM PROTOCOL THERAPY AND OFF STUDY CRITERIA Regardless of reason for r emoval from study therapy, patients will be asked to have follow up safety visit within 90 days after the last vaccine . Patients who refuse to return for this visit will be asked to review any safety concerns by phone within this time period. 3.7.1 Criteria fo r removal from protocol therapy • Completion of protocol therapy. • Clinical or radiographic progression of disease Patients with serological progression alone will remain on treatment at the discretion of the PI. • Unacceptable Toxicity as defined as any serious adverse event that is unexpected relative to the known safety profile of the investigational agents in the opinion of the investigator or as de scribed in sections 3.1.1 and 3.3 • Participant requests to be withdrawn from active therapy • Investigator discretion • Positive pregnancy test Also see section 3.3 3.7.2 Off-Study Criteria • PI decides to end study • Participant requests to be withdrawn from study. Reasons for withdrawal will be docu mented. • Noncompliance with protocol guidelines (patient removed at discretion of Principal Investigator) • Death • Screen failure 3.7.3 Off Protocol Therapy and Off -Study Procedure Authorized staff must notify Central Registration Office (CRO) when a subject is taken off protocol therapy and when a subject is taken off-study. A Participant Status Updates Form from the web site (http://home.ccr.cancer.gov/intra/eligibility/welcome.htm ) main page must be completed and sent via encrypted email to: NCI Central Registration Office ncicentralregistration - l@ma il.nih.gov . Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 38 4 CONCOMITANT MEDICATI ONS/MEASURES Subjects must inform the investigators of the current or planned use or all other medications during the study (including prescription medications, vitamins, herbal and nutritional supplements, and over-the-counter medications). For the administration of ETBX vaccines, antiemetics, stool softeners , and antidiarrheal agents may be administered as required, but are not anticipated to be needed and should not be used prophylactically on the first cycle. The selection of the specific antiemetic regimen is at th e discretion of the treating physician. Antiemetic regimens will not include steroids. Other supportive care with blood components, antibiotics, analgesics, general medical therapy, etc., will be delivered as required. Any patients taking antibiotics for infection must complete that course of therapy and be free of evidence of further infection before receiving any dose of vaccine. Use of prophylactic antibiotics is allowed. Concurrent systemic corticosteroid use (daily or every other day f or continued use > 14 days) should be avoided within 28 days before the first planned dose of ETBX vaccines. Use of physiologic doses of systemic steroids, inhaled steroids, nasal sprays, and topical creams for small body areas is allowed. Symptomatic ane mia should be treated with appropriate red blood cell or erythropoietin support. Thrombocytopenia should be treated conservatively. In the absence of bleeding or a planned invasive procedure, platelet transfusions should be given for a platelet count belo w 10,000/mm3. If invasive procedures are planned or the patient develops bleeding, platelet transfusions should be administered in accordance with the standard of practice, usually maintaining a platelet count of > 50,000/mm3. 4.1 CONCURRENT MEDICATIONS /INTER VENTIONS 4.1.1 Anticancer Therapy If a subject requires additional systemic anticancer treatment with the exception of treatments described in section 3.1 then study treatment must be discontinued. Local intervention is discouraged unless medically unavoidable. Subjects receiving local intervention ( e.g., palliative radiation) are allowed to continue to receive study treatment at the investigator’s discretion. 4.1.2 Other Medications Subjects must be instructed to inform the investigators of the current or planned use or all other medications during the study (including prescription medications, over -the-counter medications, vitamins and herbal and nutritional supplem ents). It is the responsibility of the investigator to ensure that details regarding all medications are documented. Bisphosphonates started prior to screening activities or initiated during the course of the study to control bone pain may be used with cau tion. Colony stimulating factors ( e.g., erythropoietin and granulocyte colony -stimulating factors) administered as dictated by safety purposes are acceptable while the subject is enrolled on study. Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 39 Pain medications administered as dictated by standard pra ctice are acceptable while the patient is enrolled on the study. No concurrent investigational agents are permitted . 5 BIOSPECIMEN COLLECTI ON 5.1 CORRELATIVE STUDIES FOR RESEARCH /PHARMACOKINETIC STUDIES 5.1.1 Peripheral Blood Collection Subjects will have approximate ly 70 mL of peripheral blood drawn to evaluate the study drug’s effect on the immune response at specific time points during the study and/or after a specifi ed injection . Blood draws will be done at baseline and prior to each injection (baseline, week6,14, 30 and EOT ). 5.1.2 Samples Collected Six10-mL green top sodium heparin tubes for PBMC samples and one 8 -mL serum -separating tube for serum samples will be drawn. Samples will be picked up by courier and delivered to the Leidos Biomedical Research faci lity in Frederick, MD. . 5.1.3 Sample Processing Blood samples will be processed at: Leidos Biomedical Research Attn: Dr. Ludmila Krymskaya /Theresa Burks 1050 Boyles Street Bldg. 496/Room 121 Frederick, MD 21702 5.1.4 Sample Tracking, and Disposition Samples will be ordered in CRIS and tracked through a Clinical Trial Data Management system . Should a CRIS screen not be available, the CRIS downtime procedures will be followed . Samples will not be sent outside the National Institutes for Health (NIH) w ithout IRB notification and an executed material transfer agreement. 5.1.5 Pharmacodynamic Sample Management and Storage All data associated with the patient samples is protected by using a secure database. All Clinical Support Laboratory Staff receive annual tr aining in maintaining records’ confidentiality. All samples drawn at the NIH Clinical Center will be transported to the Clinical Support Laboratory at the Frederick National Laboratory for Cancer Research by couriers. Samples will be tracked and managed by Central Repository database, where there is no link to personal identifiable information. All samples will be stored in either a -80ºC freezer or vapor phase liquid nitrogen. These freezers are located at NCI Frederick Central Repository in Frederick, Maryland. NCI Frederick Central Repositories (managed under a subcontract) store, among other things, biological specimens in support of NIH clinical studies. All specimens are stored in secure, Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 40 limited -access facilities with sufficient security, backup, and emergency support capability and monitoring to ensure long -term integrity of the specimens for research. Specimens are stored in accordance wi th applicable HHS and FDA Protection of Human Subjects Regulations in accordance with the subcontractor ’s Federal -wide Assurance. The subcontractor ’s role limited to clinical research databases and repositories containing patient specimens. The subcontractor does not conduct or have any vested interest in research on human subjects, but does provide services and support the efforts of its customers, many of which are involved in research on human subjects. The subcontractor ’s IRB reviews policies and procedures for labeling, data collection and storage, access, and security. The IRB will review protection of p rivacy issues prior to acceptance of any new work and in the event of change impacting privacy issues in existing work. It is the intent and purpose of the subcontractor to accept only de -identified samples and sample information. To the limit of our abil ity, every effort will be made to ensure that protected information is not sent electronically or by hard copy or on vial labels. Sample data is stored in the BioSpecimen Inventory System II (BSI). This inventory tracking system is used to manage the stor age and retrieval of specimens as well as to maintain specimen data. BSI is designed for controlled, concurrent access. It provides a real -time, multi -user environment for tracking millions of specimens. The system controls how and in what order database u pdates and searches are performed. This control prevents deadlocks and race conditions. For security, BSI has user password access, 3 types of user access levels, and 36 user permissions (levels of access) that can be set to control access to the system fu nctions. BSI provides audit tracking for processes that are done to specimens including shipping, returning to inventory, aliquoting, thawing, additives, and other processes. BSI tracks the ancestry of specimens as they are aliquoted, as well as discrepanc ies and discrepancy resolution for specimens received by the repository. If a specimen goes out of the inventory, the system maintains data associated with the withdrawal request. Vials are labeled with a unique BSI ID which is printed in both eye -readable and bar -coded format. No patient -specific information is encoded in this ID. Investigators are granted view, input, and withdrawal authority only for their specimens. They may not view specimen data or access specimens for which they have not been author ized. Access to specimen storage is confined to repository staff. Visitors to the repositories are escorted by repository staff at all times. 5.1.6 Sample Analysis Immune assessments will be performed at the Laboratory of Tumor Immunology and Biology at the NCI’ s Center for Cancer Research (CCR) and include flow cytometry -based and serum assays. Analyses of PBMCs: 1. Pre- and post -therapy PBMCs, separated by Ficoll -Hypaque density gradient separation, will be analyzed for antigen -specific immune responses using an intracellular cytokine staining assay. PBMCs will be stimulated in vitro with overlapping 15 -mer peptide p ools encoding the tumor -associated antigens CEA, MUC1, and Brachyury. Control peptide pools will involve the use of human leukocyte antigen peptide as a negative control and CEFT peptide mix as a positive control. CEFT is a mixture of peptides of CMV, Epst ein- Barr virus, influenza, and tetanus toxin. Post -stimulation analyses of CD4 and CD8 T cells Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 41 will involve the production of IFN -γ, IL-2, tumor necrosis factor, and CD107a. If sufficient PBMCs are available, assays may also be performed for the developmen t of T cells to other tumor -associated antigens. A detailed description of this assay has been previously reported (35). 2. If sufficient PBMCs are available, PBMCs from selected subjects may be analyzed for changes in standard immune cell types (CD4 and CD8 T cells, natural killer [NK] cells, regulatory T cells [Tregs], myeloid -derived suppressor cells [MDSCs], and dendritic c ells) as well as 123 immune cell subsets, as described elsewhere (74), and for function of specific immune cell subsets, including CD4 and CD8 T cells, NK cells, Tregs, and MDSCs. Analyses o f soluble factors: Sera will be analyzed pre - and post -therapy for the following soluble factors: sCD27, sCD40 ligand, and selected patients may be analyzed for cytokines and antibodies to human tumor antigens. For selected patients, sera may be analyzed f or antibodies to CEA, MUC1, or Brachyury and adenovirus . Additional assays: Blood samples may be used for additional research studies, which may include phenotypic and functional analysis of immune -cell subsets and analyses for cytokines (IFN -γ, IL -10, IL-12, IL -2, IL-4, etc.), chemokines, antibodies, tumor -associated antigens, and/or other markers. In addition, assays for antibody or neutralizing antibody titer to adenovirus (type 5) and cell -mediated immunity to adenoviral antigens may be assessed. Samples will be tracked according to Section 5.1.4 . 5.1.7 Protocol Completion/Sample Destruction All specimens obtained in the protocol are used as defined in the protocol. Any specimens remaining at the completion of the protocol will be stored in the conditions described above. The study will remain open as long as sample or data analysis continues. Samples will be stored until they are no longer of scientific value or until a subject withdraws consent for their continued use, at which time they will be destroyed. Once primary research objectives for the protocol are achieved, intramural researchers can request access to remaining samples, provided they have an IRB-approved protocol and subject consent or an exemption from the Office of Human Research Protections . The Investigator will report any loss or destruction of samples to the NCI IRB as soon as he/she is made aware of such loss. The Investigator will report destroyed samples to the IRB if samples become unsalvageable because of environmental factors such as a broken freezer or lack of dry ice in a shipping container, or if a subject withdraws consent. Samples will also be reported as lost if they are lost in transit between facilities or misplaced by a researcher. Freezer problems or lost samples will also be reported to the IRB, the NCI Clinical Director, and the office of the NCI CCR. Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 42 6 DATA CO LLECTION AND EVALUAT ION 6.1 DATA COLLECTION Data will be entered in C3D. The PI will be responsible for overseeing entry of data into an in - house password protected electronic system and ensuring data accuracy, consistency and timeliness. The principal invest igator, associate investigators/research nurses and/or a contracted data manager will assist with the data management efforts. All data obtained during the conduct of the protocol will be kept in secure network drives or in approved alternative sites that comply with NIH security standards. Primary and final analyzed data will have identifiers so that research data can be attributed to an individual human subject participant. All adverse events, including clinically significant abnormal findings on laboratory evaluations, regardless of severity, will be followed until return to baseline or stabilization of event. Patients will be followed for adverse events for a minimum of 30 days after removal from study treatment or until off -study, whichever come s first. An abnormal laboratory value will be recorded in the database as an AE only if the laboratory abnormality is characterized by any of the following: • Results in discontinuation from the study • Is associated with clinical signs or symptoms • Requ ires treatment or any other therapeutic intervention • Is associated with death or another serious adverse event, including hospitalization. • Is judged by the Investigator to be of significant clinical impact • If any abnormal laboratory result is consid ered clinically significant, the investigator will provide details about the action taken with respect to the test drug and about the patient’s outcome. End of study procedures: Data will be stored according to HHS, FDA regulations, and NIH Intramural Reco rds Retention Schedule as applicable. Loss or destruction of data: Should we become aware that a major breach in our plan to protect subject confidentiality and trial data has occur red, the IRB will be notified. 6.2 DATA SHARING PLANS 6.2.1 Human Data Sharing Plan What data will be shared? I will share human data generated in this research for future research as follows: • De-identified data in an NIH -funded or approved public repository. • De-identified data in BTRIS (automatic for activities in the Clinical Center) • De-identified or identified data with approved outside collaborators under appropriate agreements. How and where will the data be shared? Data will be shared through: Abbreviated Title: Ad5 -based combination vaccines Version Date: 05.16.2018 43 • An NIH -funded or approved public repository. Insert name or names:_ clinicaltrials.gov _. • BTRIS (automatic for activities in the Clinical Center) • Approved outside collaborators under appropriate individual agreements. • Publication and/or public presentations. When will the data be shared? • Before publication.