Jessica Reichow

Topical Imiquimod Plus Nab-paclitaxel for Breast Cancer Cutaneous Metastases: A Phase 2 Clinical Trial.

Importance Salvage chemotherapy for recurrent chest wall lesions in breast cancer results in response rates of 20% to 30%. Preclinical studies showed significant disease regression could be induced in murine chest wall mammary cancers with a topical toll-like receptor (TLR)-7 agonist, imiquimod. Objective To evaluate the safety and objective response rate (ORR) of imiquimod in combination with systemic albumin bound paclitaxel in treatment-refractory breast cancer of the chest wall. Design, Setting, and Particpants A single arm phase 2 clinical trial of 15 patients with breast cancer previously treated in an academic medical center setting between 2009 and 2012 for chest wall disease that had recurred. Interventions Imiquimod cream, 5%, was applied topically to a designated target lesion once per day for 4 consecutive days on days 1 through 4, 8 through 11, 15 through 18, and 22 through 25 of a 28-day cycle, for 12 weeks. Albumin bound paclitaxel, 100 mg/m2, was given intravenously on days 1, 8, and 15, and repeated every 28 days over the 12-week period. Main Outcomes and Measures The primary endpoint was safety and ORR. Secondary endpoints included the generation of tumor-infiltrating lymphocytes and modulation of immune cell populations. Results The median age at baseline of the 15 study participants was 54 years (range, 46-92 years). Fourteen patients were evaluable. Combination therapy was associated with low-grade toxic effects. Of 358 adverse events 330 (92%) were grades 1 and 2. Five (36%) patients achieved a compete response and another 5 (36%) were partial responders for an overall response rate of 72% (10 of 14). The response duration was limited. Pretreatment levels of programmed death-1 (PD-1)+ peripheral blood T cells (PD-1+ cluster of differentiation [CD]4+; 95% CI, 2.68-6.63; P < .001 and PD-1+CD8+; 95% CI, 1.13-8.35; P = .01) and monocytic myeloid derived suppressor cells (mMDSC) (95% CI, 3.62-12.74; P = .001) greater than controls predicted suboptimal clinical response. Conclusions and Relevance Chemoimmunomodulation with a TLR-7 agonist and albumin bound paclitaxel is effective in inducing disease regression in treatment-refractory breast cancer chest wall metastases but responses are short-lived. Preexisting levels of cells indicating either T-cell exhaustion or systemic immunosuppression may be markers of selection for responsive patients. Trial Registration clinicaltrials.gov Identifier: NCT00821964

Abstract OT3-1-02: Phase II randomized study of combination immunotherapy with or without Polysaccharide Krestin (PSK®) concurrently with a HER2 ICD peptide-based vaccine and trastuzumab in patients with stage IV breast cancer

Background: Endogenous immunity in patients with HER2+ metastatic breast cancer (MBC) is likely dampened by an immune-suppressive tumor microenvironment and not sufficient to control tumor growth. Thus, most patients have disease relapse after achieving complete remission with standard therapies. Immunomodulation directed at enhanced stimulation of tumor specific immunity could result in immunologic eradication of residual HER2+ tumor cells and prevent BC relapse. We have shown PSK to be a potent TLR-2 agonist that stimulates both innate and adaptive immunity in a BC mouse model. Additionally, we have shown combination immunotherapy with HER2 peptide vaccines and trastuzumab (TRAZ) to be safe and able to elicit HER2 specific Th1 immunity and epitope spreading (ES) which has been associated with survival in vaccinated patients. Lastly, decreased serum TGF-β elicited by HER2 vaccination correlates with Th1 ES and may serve as a biomarker to predict cancer vaccine efficacy. We hypothesize that PSK, when given with TRAZ can augment vaccine induced HER2 specific TH1 immunity and prevent disease relapse in patients with optimally treated HER2+ MBC. Trial design: Phase II randomized two-arm clinical trial. Patients will be enrolled and randomly assigned in equal numbers to 1 of 2 arms (15 patients/arm) as follows: Arm 1:HER2 ICD vaccine, TRAZ and placebo or Arm 2:HER2 ICD vaccine, TRAZ and PSK. All patients will receive 3 monthly HER2 ICD vaccines plus TRAZ and 4 months of concomitant PSK or placebo. Serial blood draws for immunologic monitoring will be done. Eligibility criteria: Patients with Stage IV HER2+ BC who have been treated with definitive therapy and are: (1) without evidence of disease or have stable-bone only disease, (2) receiving TRAZ monotherapy, and (3) without clinically significant autoimmune disease. Patients must have normal LVEF per MUGA scan or echocardiogram. Aims: (1) Evaluate safety of PSK when given with a HER2 vaccine and TRAZ (2) Evaluate the effect of PSK on serum TGF-β levels when given with a HER2 vaccine and TRAZ and (3) Evaluate the effect of PSK on intermolecular ES when given with a HER2 vaccine and TRAZ. A secondary objective is to evaluate progression free survival (PFS) and overall survival (OS). Statistical methods: (1) Toxicity will be determined by clinical and chemical parameters and grading will be done per CTEP CTCAE 4.0.; (2) Evaluation of TGF-β levels, pre and post-PSK treatment will be assessed with linear regression models; and analysis of multiple post-baseline measurements will be performed using generalized estimating equations; (3) A positive antigen-specific immune response will be defined as a precursor frequency >1:20,000 antigen-specific peripheral blood mononuclear cells. Differences in the levels of HER2 immunity will be evaluated between arms using a two-tailed T test. The degree of ES in each arm will be evaluated with generalized linear modeling; (4) Large differences in PFS and OS observed between groups will be noted and described. Target accrual: 30 patients. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr OT3-1-02.

Phase II trial of albumin-bound paclitaxel and granulocyte macrophage colony-stimulating factor as an immune modulator in recurrent platinum resistant ovarian cancer

BACKGROUND Granulocyte macrophage colony-stimulating factor (GM-CSF) stimulates immunity via recruitment of antigen presenting cells and tumor specific T-cell stimulation. Albumin-bound paclitaxel (nab-paclitaxel) followed by GM-CSF may enhance antitumor responses and prolong remissions in ovarian cancer. Immune phenotypes present before treatment may identify responders to chemo-immunotherapy. METHODS Recurrent platinum-resistant ovarian, peritoneal, or fallopian tube cancer patients received nab-paclitaxel, 100mg/m2 days 1, 8, 15 followed by GM-CSF 250μg days 16-26 every 28days for 6 planned cycles. The primary endpoint was remission duration compared to immediate prior remission. Peripheral blood was evaluated by flow cytometry and interferon-γ ELISPOT. RESULTS Twenty-one patients were enrolled. Six patients (29%) achieved a biochemical complete response and 9 (43%) a partial response for an overall response rate of 72%. Median time to progression was 4months and 10% of patients achieved longer remissions than the immediate prior regimen. Median overall survival (OS) was 16.8months. Fewer myeloid derived suppressor cells (MDSC) at enrollment significantly associated with complete response (p=0.05). T-cell responses to IGF1R-p1332-1346 (r=0.827, p=0.0003) and IGF1R-p1242-1256 (r=0.850, p=0.0001) during treatment correlated with time to progression. CONCLUSIONS Nab-paclitaxel combined with GM-CSF demonstrated biochemical responses in a majority of patients, although responses were not sustained. This combination did not demonstrate an advantage in OS over prior studies of nab-paclitaxel monotherapy. Agents that modulate MDSC should be studied as potential adjuvants to therapy. Strategies to expand T cells recognizing tumor-associated antigens biologically significant in ovarian cancer should also continue to be investigated.

Abstract NTOC-097: VACCINATION TARGETING INSULIN–LIKE GROWTH FACTOR BINDING PROTEIN–2 (IGFBP–2) IN ADVANCED OVARIAN CANCER: SAFETY, IMMUNOGENICITY, AND SURVEILLANCE, EPIDEMIOLOGY, AND END RESULTS (SEER) COMPARISON

BACKGROUND: Immunization against self-antigens can induce regulatory responses that inhibit desirable Type 1 antitumor immune responses. Deletion of epitopes that favor a regulatory phenotype may improve the efficacy of therapeutic vaccines. We have developed a novel IGFBP-2 targeting DNA plasmid vaccine that selectively induces Type 1 immunity. IGFBP-2 regulates invasiveness and metastases in ovarian cancer. Eradication of ovarian cancer cells expressing IGFBP-2 through effective immunization could prevent disease relapse or metastasis. METHODS: Twenty-five patients with advanced stage or recurrent ovarian cancer treated to complete remission after primary or salvage therapy received 3 monthly doses of an IGFBP-2 DNA vaccine in a single-arm, non-randomized study. ELISPOT and flow cytometry were used to characterize antigen specific T-cell responses. Serum antibodies were measured using ELISA and Western blot. The SEER database was reviewed to identify women diagnosed between 2006 and 2012 matched for age, year of diagnosis and stage of diagnosis. The difference between dates of diagnosis and enrollment (lead time) was calculated for each patient receiving vaccine. Only SEER patients who survived at least as long as the lead time of their matches plus an additional 6 months were kept for analysis. In cases where this resulted in no SEER matched patients, unmatched vaccine patients were excluded. Overall survival (OS) was analyzed using Cox models and the Kaplan-Meier method. RESULTS: 206 adverse events (AE) were recorded. Fatigue (12%) and injection site reactions (12%) were the most common. 97% of AE were grades 1-2, 3% grade 3, and no grades 4 or 5. In preliminary immune analysis (16 patients), IGFBP-2 specific T-cell precursor frequencies are significantly elevated over baseline levels at 4 (p CONCLUSIONS: IGFBP-2 Th1 selective immunization is well tolerated, generates significant Type I immunity, and may demonstrate clinical efficacy. Citation Format: John B. Liao, Denise L. Cecil, Yushe Dang, Kelsey K. Baker, Kelsie J. Ovenell, Jessica Reichow, Stephanie Parker, Doreen M. Higgins, Jennifer S. Childs, Elizabeth K. Broussard, Andrew L. Coveler, Lupe G. Salazar, Barbara A. Goff, Mary W. Redman, Mary L. Disis. VACCINATION TARGETING INSULIN–LIKE GROWTH FACTOR BINDING PROTEIN–2 (IGFBP–2) IN ADVANCED OVARIAN CANCER: SAFETY, IMMUNOGENICITY, AND SURVEILLANCE, EPIDEMIOLOGY, AND END RESULTS (SEER) COMPARISON [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-097.

Abstract P2-11-05: Overall survival in inflammatory breast cancer patients receiving Her-2 Neu directed tumor vaccine therapy: Matched comparison with SEER registry patients

Background Patients with inflammatory breast cancer (IBC) have a poor prognosis, primarily due to distant dissemination. Additionally, IBC patients have an increased rate of HER2 overexpression when compared to patients with non-inflammatory breast cancer. The forms the rationale for HER2 directed tumor vaccine therapy in these patients. The purpose of this study was to examine overall survival in IBC patients receiving HER2 directed tumor vaccine therapy when compared with matched control patients from the SEER Registry. Methods Patients with diagnosis of Stage III or IV HER2 positive IBC having completed standard initial therapy and without evidence of disease received HER2 vaccinations after being enrolled on 5 prospective clinical trials. Overall survival data were pooled and analyzed. A control group of matched IBC patients were identified by querying the SEER database from 1997-2011. The control group was identified as any individual in the database with a code for IBC. A secondary analysis comparing survival in HER2 positive IBC vs HER2 negative IBC patients was performed by querying the SEER database from 2010 onwards, the time point when the HER2 status was coded in the database. Propensity score adjustment were made to the control group to account for any imbalances between groups in measured covariates such as stage, race, age, sex, and era of enrollment and the time interval from diagnosis to enrollment on vaccine trial (median ∼2 years). Results A total of 37 IBC patients received HER2 directed vaccine therapy and 676 patients were identified for the SEER control group; Stage at enrollment: stage IIIB: 30 patients in the vaccine group and 639 patients in the control group; stage IIIC: 1 patient in the vaccine group and 15 patients in the control group; stage IV 6 patients in the vaccine group and 22 in the control group. The median survival of the overall population was 112 months for the vaccine group and 47 months for the control group (p=0.04). After using propensity scores to adjust the control for imbalances in measured covariates, the median survival for the overall population was 112 months for the vaccine group and 37 months for the control group (p=0.03). There was no difference in survival between HER2 positive and HER2 negative IBC patients in the control group (p=0.6). Conclusion These results demonstrate promising overall survival in HER2 positive IBC patients receiving HER2 directed vaccine therapy after initial therapy. Propensity matching was performed to adjust for imbalances in measured covariates and resulted in a modest decrease in survival of the control group after adjustment, suggesting that the vaccine trial group had relatively unfavorable pre-treatment characteristics. Despite these unfavorable characteristics, patients receiving vaccine had a median survival of 112 months. These results must be further confirmed in a prospective randomized trial. Citation Format: Rengan R, Baker K, Salazar L, Childs J, Higgins D, Redman M, Reichow J, Disis ML. Overall survival in inflammatory breast cancer patients receiving Her-2 Neu directed tumor vaccine therapy: Matched comparison with SEER registry patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-11-05.

Abstract 3135: HER2 specific adoptive T cells shown to localize and infiltrate all sites of disease using combined SPECT and PET imaging

Adoptive T cell therapy has been shown to stimulate anti-tumor response in multiple cancers, however these responses have not been robust or durable. In previous studies, indium-111 labeled T cells have functioned normally but were either not present at all sites of metastatic disease or not able to infiltrate the sites of disease. In breast cancer, adoptive HER2 targeted T cells were unable to penetrate visceral metastases. Previous work in our laboratory has demonstrated that HER2 vaccine-primed autologous adoptive T cells were safe and well tolerated. In this phase I study, HER2 positive breast cancer patients received three HER2 peptide vaccines before plasmapheresis and ex-vivo expansion of HER2 specific autologous T cells to evaluate the immune and clinical response to adoptive T cell therapy in breast cancer. In one patient, the trafficking of indium-111 labeled T cells was also evaluated using SPECT and PET imaging. An aliquot of 1×107 expanded T cells were labeled with 300 uCi of indium-111 and given with the third T cell infusion. Prior in vitro studies had demonstrated that labeled HER2 expanded T had similar viability (at 24 hours 97±1% viability with unlabeled cells and 90.9±1.1% viability with 480 uCI labeled cells) and interferon gamma release (263±8 pg/mL released in unlabeled cells and 208.5±11 released in 480 uCi labeled cells) as unlabeled HER2 expanded T cells when stimulated by IL2. SPECT imaging demonstrated that the T cells trafficked to all the metastatic sites of disease by 24 hours and completely infiltrated the tumor. The patient studied had metastases to her skull, left axilla, sternum, bilateral proximal humeri, and sacrum. The corrected indium-111 uptake at 24 hours varied from 2.27 counts per pixel in the R proximal humerus to 6.28 counts per pixel in the R sacrum and remained elevated at 48 hours (for example a continued 6.9 counts/pixel signal at the R sacrum). Concurrent PET CT imaging demonstrated FDG flare at 48 hours at all sites of metastatic disease including a 1.3 fold increase in the R proximal humerus and a 1.3 fold increase in the L proximal humerus signal over baseline scans. This increased FDG uptake had resolved 1 month after therapy. After this study, the patient had stable disease for 18 months. She had a robust response to each T cell infusion and the booster vaccines, including fevers, headaches, and increased pain at the sites of metastatic disease. These symptoms have been associated with a disease specific T cell response. This study demonstrates by a novel method of concurrent SPECT and PET imaging that the ex-vivo expanded HER2 specific T cells were able to traffic to and fully infiltrate all sites of metastatic disease causing an acute FDG-PET flare and prolonged stable disease in a HER2 positive breast cancer patient with bone-only metastatic disease. Further studies are now needed to confirm if this imaging method can be used universally in adoptive T cell studies. Citation Format: Sasha E. Stanton, Janet Eary, Edmond Marzbani, David Mankoff, Lupe Salazar, Doreen Higgins, Jessica Reichow, Yushe Dang, Mary L. Disis. HER2 specific adoptive T cells shown to localize and infiltrate all sites of disease using combined SPECT and PET imaging. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3135. doi:10.1158/1538-7445.AM2015-3135

Persistent immunity after therapeutic vaccination targeting HER2/neu correlated with post-vaccination magnitude of interferon gamma Elispot responses

Meeting abstracts As therapeutic vaccination approaches are explored for cancer therapy, there is a need to identify milestones that can predict the successful induction of sustained immunity. We examined whether the magnitude of immune responses seen prior to vaccination or at the completion of a

Abstract P2-15-02: The efficacy of recruitment and retention strategies for research subjects in an early phase investigator-initiated breast cancer trial.

Background: One of the biggest challenges faced by investigators is the implementation of effective strategies to improve the recruitment and retention of research participants. This is especially true for investigator-initiated, federally funded (e.g. NIH and DOD), early phase clinical trials that involve the treatment of serious diseases such as metastatic breast cancer (MBC). These studies may face additional barriers to participation since patients have often already undergone maximal treatment and are usually not in a financial position that for allows the travel and lodging necessary to receive further investigative treatment. Moreover, efficacy and toxicology in early phase clinical trials are unknown. Thus, when study budget constraints do not allow monetary incentives to participation, it is difficult to provide motivation for patients to enroll and remain adherent to the protocol requirements. However, many MBC patients are motivated to join clinical trials for altruistic purposes alone, and evidence supports that the researcher-patient relationship may be the most important factor in clinical trial participation. Recognizing that many patients are willing to participate if provided the appropriate resources despite limited monetary incentives, we developed a system to improve patient recruitment and retention to our studies, which are primarily federally funded. We report here on the strategies developed and used by our group to recruit and retain patients in a federally-funded investigator-initiated phase I/II vaccine study in MBC patients. Methods: This study was funded by the NIH/NCI and involved infusion of HER2 specific T cells in HER2+ MBC patients after completing in vivo priming with a HER2 vaccine. It required 11 visits to Seattle, Washington. Working with agencies that offer free services to patients enrolled in clinical trials, a list of available resources was compiled and a visit flowchart with specific information on travel and lodging resources (e.g. Angel Flights and ACS sponsorship), local transportation and entertainment was developed. During screening, patients were given the list of resources and trial information. An email system was used to quickly communicate and follow-up with patients. Eligible patients were given the visit flowchart to help with their planning of study visits. An enrollment packet was provided at the first visit with a calendar to keep track of the visit schedule. Coordination of care between the patient9s primary oncologist and the research staff was maintained throughout the study. Results: 17 of 19 patients enrolled were not from Washington State. Two out-of-state patients withdrew early from the trial for reasons unrelated to disease progression or toxicity; one subject completed 8 visits and enrolled in another study and the other completed 2 visits and discontinued the trial to resume chemotherapy. Conclusion: We have developed a successful system to enroll and retain patients in a trial requiring multiple study visits. Development and implementation of site-specific standard procedures are critical to improve study participation and retention, especially when patients receive no financial benefit. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-15-02.