Research Funding

Year Funded: 2013

Characterization of Potential Antigens for Immunization Against Colon Cancer, and Testing Multivalent Tumor Vaccines in Min Mice

Grant Number: OUHSC-RS2O122 159-02

The successful identification of novel tumor antigens is critical to cancer immunotherapy and its potential use in cancer prevention. Colon cancer is the third most common cancer both in men and women and is the second most common cause of cancer death. Mouse models that mimic human disease provide a unique tool for tumor antigen discovery. In the past, multiple groups have used a variety of rodent models of colon cancer (Min, AOM Mouse, AOM Rat) to examine colon cancer development, potential agents that can prevent cancer development (e.g. NSAIDs) etc. In a more limited number of cases, they have used these models to examine the role of the immune response and even to make initial examination of immunization against colon cancer.

There are a variety of approaches towards deciding upon antigens which might be employed to develop a vaccine. In the rare cases where there is a highly over expressed protein that appears to be directly associated with tumor development e.g. Neu in Neu over expressing tumors this appears to be a promising approach both in animal models and clinically. A second is to pick antigens which are over expressed in both model and human tumors and which appear to be associated with the tumor process. It is this latter approach that will be employed here.

The specific animal model that will be employed here is a modified Min model. Although the standard Min model, on a C57BL6 background, has proven effective in screening potential chemopreventive agent it has a major limitation for vaccine work. The adenomas arise quite rapidly so that an animal will already have multiple adenomas at the time that immunization is initiated. To overcome this, the Contractor shall employ an F1 Min mouse (C57BlxAKR). In this case tumors come up more slowly which allows for initial immunization of animals with minimal tumor burden. Furthermore, animals survive long enough that they develop adenocarcinomas in the small intestine and a significant number of lesions of the colon unlike the standard Min mice.

Enhancing innate immunity by combining PSK with docetaxel in prostate cancer

Grant Number: H34B12-07 Prime: 1U19AT006028

Bastyr University collaboration for the University of Washington Immunologic

Monitoring Lab to accomplish the scientific aim of immune monitoring of blood samples for the

Bastyr/University of Washington PSK® prostate cancer clinical trial.

Year Funded: 2012

Optimized ex vivo expansion of anti-tumor Th1 and Tc1 for adoptive immunotherapy

Grant Number: CA136632

Recent clinical trials have demonstrated that advanced melanoma can be treated with a combination of nonmyeloablative chemotherapy and autologous adoptive T cell immunotherapy (AIT) to achieve a 50% clinical response rate. Such clinical responses are not, however, consistently durable. Cytotoxic T cells (CTL) are shortlived in the absence of T cell help (Th), with limited persistence after infusion. Furthermore, antigen (Ag) negative variants develop after infusion of CD8+ predominant tumor specific T cell lines, suggesting tumors readily escape from focused therapy. Our goal is to optimize and extend AIT to advanced breast cancer by providing functionally characterized tumor Ag-specific CD4+ Th1 cells as a central component of the infused product. Tumor Ag-specific CD4+ Th1 cells can home to tumor and secrete inflammatory cytokines, modulating the microenvironment to enhance the function of local antigen presenting cells (APC). The resultant increased processing of endogenous tumor cells results in epitope spreading. By providing a robust CD4+ Th1 immune response, tumor Ag-specific CD8+ T cells will be elicited, and the response generated will be long lived. We aim to promote tumor-specific, epitope spreading, Th1-type CD4+ responses as an essential component of effective AIT. This strategy has been validated by our own vaccine trials for Stage III/IV patients with HER-2/neu (HER2)-overexpressing breast cancer. Patients vaccinated with MHC Class II-binding HER2-derived peptides evidenced a significant survival advantage if they achieved not only enhanced CD4+ T cell responses to vaccine peptides, but also T cell responses to additional HER2 epitopes expressed by their tumors but absent from the vaccine. In animal studies we observe that the inclusion of autologous dendritic cells (DCs) during T cell culture can markedly improve T cell therapeutic performance at the time of re-infusion. Moreover, appropriately activated DCs can promote the expansion of T cells with higher functional avidity, including CD8+ cytolytic T cells that can directly lyse MHC-restricted tumors. We propose to develop a clinically relevant method to expand HER2 specific T cells ex vivo, using optimally activated autologous DC generated simultaneously in the same cultures as the T cells to be activated. We will also assess whether these culture methods increase epitope spreading.

Year Funded: 2011

Adoptive transfer of tumor specific Th1 cells derived from vaccine-primed patients achieved clinical benefits

Grant Number: Rivkin

Adoptive immunotherapy (AIT) can induce cancer regression but rarely results in cure. We have infused HER2 specific Th1 cells in breast cancer patients and a 50% overall objective response was observed. We hypothesize that polyfunctional tumor antigen-specific Th1/Th17 cells have enhanced therapeutic efficacy. We will determine the optimal conditions to expand the multifunctional T cells ex vivo and examine the efficacy and safety of infusion of IGFBP2 specific multifunctional T cells in a mouse model of human ovarian cancer. This study will lead to a phase I study of AIT in ovarian cancer after priming with an IGFBP2 vaccine.

Phase I-II Study of HER2 Vaccination with Ampligen as Adjuvant in Optimally Treated Breast Cancer Patients

Grant Number: Hemispherx

Due to recent advances in the conventional treatment of breast cancer (BC), BC patients are frequently able to achieve a state of complete remission with current primary therapies. However, approximately 30% of patients will eventually have recurrent disease, with the majority of recurrences being to metastatic sites; suggesting that relapse is due to residual microscopic disease. This is especially true in HER2+ BC patients who have worse disease-free and overall survival when compared to similar HER2 negative patients. One approach to the eradication of residual subclinical disease in this high-risk population is the development of tumor vaccines that target HER2, an immunogenic biologically relevant protein. Generation of vaccine-induced HER2-specific Type I inflammatory CD4+ T helper immunity (Th1) could result in immunologic eradication of residual HER2+ tumor cells and potentially prevent disease relapse or tumor spread.

We have demonstrated the ability to induce HER2-specific Th1 immunity and epitope spreading in breast cancer patients vaccinated with HER2 peptide-based vaccines given intradermally in combination with GM-CSF as adjuvant. Given the importance of Th1 immunity and its role in generating antigen-specific inflammatory responses, it is critical to investigate other vaccine adjuvants that can augment both development and magnitude of Th1 HER2-specific immunity. One such potential adjuvant is Ampligen [poly(I) • poly(C12,U)] which mediates its immunomodulating activity exclusively as a TLR-3 agonist.

We hypothesize HER2 peptide-based vaccination given in combination with GM-CSF and Ampligen can induce a higher incidence and magnitude of HER2-specific Th1 immunity than HER2 vaccination with GM-CSF alone. The aims of this study are to: (1) choose the most promising of 4 different Ampligen doses as an adjuvant to HER2 vaccination, with respect to toxicity and immune response and (2) determine, using the optimal Ampligen dose from Aim 1, whether Ampligen increases the incidence and magnitude of vaccine immune response as compared to the standard GM-CSF adjuvant strategy.

Characterization Of Potential Antigens For Immunization And Testing Of A Vaccine Against Colon/Intestinal Cancer In Min Mice

Grant Number: HHSN261-200433001C

The successful identification of novel tumor antigens is critical to cancer immunotherapy and its potential use in cancer prevention. Colon cancer is the third most common cancer both in men and women and is the second most common cause of cancer death. Mouse models that mimic human disease provide a unique tool for tumor antigen discovery. In the past, multiple groups have used a variety of rodent models of colon cancer (Min, AOM Mouse, AOM Rat) to examine colon cancer development, potential agents that can prevent cancer development etc. In a more limited number of cases they have used these models to examine the role of the immune response and even to make initial examination of immunization against colon cancer. Three particular aspects of colon cancer and the relevant rodent cancer models make this particularly appealing. First the preponderance of colon tumors and human FAP are driven by alterations in the WNT pathway most typically truncating mutations in the APC gene. Thus, the driving process in most human colon cancers and in the rodent tumor models listed above are alterations in the WNT pathway. Probably reflecting this common pathway alteration microarray analyses of normal mucosa and lesions from humans as compared to mice show significant overlap with regards to gene expression pathways which are altered as well as with regards to various specific genes whose expression is altered between tumors and normal mucosa. This is potentially important since certain of these proteins may prove to be antigens which are potentially useful for developing a vaccine. Potentially the third aspect of colon cancer that makes it particularly appealing is that there is some significant literature implying that colon cancers are often associated with a significant immunologic infiltrate and that the extent of this infiltrate may have significant effects on the prognosis of the colon cancer.

There are a variety of approaches towards deciding upon antigens which might be employed to develop a vaccine. In the rare cases where there is a highly over expressed protein that appears to be directly associated with tumor development e.g. Neu in Neu over expressing tumors this appears to be a promising approach both in animal models and clinically. A second is to pick antigens which are over expressed in tumors and which appear to be associated with the tumor process. More recently there has been an attempt to determine whether within the group of over expressed antigens there may be some that have elicited either an antibody response (SEREX) or a T cell mediated response in individuals with the tumor or in the case of animal models who may have rejected a tumor. The particular appeal of these antigens is that individuals have responded to these proteins and by implication it may be easier to break tolerance against these specific antigens. This specific approach has been pursued in the MMTV-Neu model in mice.

The objective of this work assignment would be to:
(1) Based on expression data determine whether expression of specific proteins, as determined by RNA when comparing colon lesions and normal mucosa, is similar in human colon tumors and certain animal models ( Min mouse).
(2) Determine whether animals bearing intestinal lesions demonstrate immunologic responses as determined by SEREX or specific T cell reactivity against relevant antigens and compare this with relevant proteins which have elicited an immune response in persons bearing colon tumors.
(3) From these proteins develop four antigens which appear amenable to development as potential antigens in a multivalent vaccine.
(4) Test these antigens for their ability to elicit an immune response in min mice.
(5) Test the potential vaccine (4 antigens vs 2 antigens and 2 antigens) to inhibit tumor formation in Min mice
(6) Test the vaccine together with a sub-ideal dose of a chemopreventive agent e.g. Celecoxib.

Vaccine to Prevent Breast Cancer

Grant Number: W81XWH-11-1-0760

Vaccination directed against common human pathogens has been one of the most successful strategies used to prevent human disease. The success of infectious disease vaccines depends on immunizing people prior to exposure to the pathogen. If we could identify immunogenic proteins important for breast cancer initiation, a vaccine could be developed to prevent breast cancer. Our group has shown that upregulation of a breast cancer-associated protein can result in that protein becoming immunogenic. This observation has allowed prospective screening of proteins that are overexpressed and associated with cancer stem cells and/or epithelial to mesenchymal transformation (EMT) as vaccine candidates. Stem cells carry the ability to initiate and maintain breast cancers and are associated with self renewal and metastatic potential, and EMT may drive the acquisition of stem cell-like properties. Eliminating cells that had upregulated proteins associated with the cancer stem cell and EMT may eliminate breast cancer. Vaccine strategies designed to elicit tumor antigen specific T helper 1 (Th1) immunity have the potential to elicit epitope spreading, concurrently stimulate antigen specific cytotoxic (CTL) CD8+ T cells, and establish immunologic memory. Immunologic memory will ensure that the tissue destructive immune response will deploy whenever the antigen is expressed in the future. We propose to develop a multiantigen polyepitope Th vaccine targeting stem cell/EMT antigens for the prevention of breast cancer.

Year Funded: 2010

PSK as Neoadjuvant for Locally Advanced Breast Cancer

Grant Number: CA138547

Locally advanced breast cancer (LABC) refers to a breast cancer that has progressed locally but has not yet clinically spread beyond the breast and regional lymph nodes. Clinical management of LABC remains challenging as the patients have a high risk for relapse. This is particularly true for patients whose tumors overexpress the HER-2/neu oncogenic protein (HER2+) and have limited expression of estrogen receptors (ER-). Neoadjuvant (pre-operative) chemotherapy followed by surgery has evolved as the standard treatment strategy for newly diagnosed LABC. Patients with pathological complete response (PCR) achieved by neoadjuvant therapy have a lower relapse rate after surgery and an improved overall survival compared to those patients with residual microscopic disease. However, with the currently available neoadjuvant therapy for HER2+/ER- patients, including chemotherapy and monoclonal antibody (mAb) therapy, PCR is achieved only in a minority of patients. Novel therapeutic strategies are required to result in complete tumor eradication. Stimulating an adaptive immune response against the tumor could provide a treatment that (1) is long lasting secondary to the development of immunologic memory, (2) has the potential to home to any tumor deposit and eradicate existing cells, and (3) has low toxicity due to the specificity of the T cell-antigen interaction. We propose to add polysaccharide Krestin (PSK), a non-toxic immunomodulator, to standard neoadjuvant therapy as a consolidation approach. PSK has long been used in Asia as an immune potentiating and anti-cancer drug. Our preliminary experiments in neu transgenic mice, a model of human HER2+/ER- LABC, have shown that PSK by itself augments tumor specific immunity and inhibits tumor growth. Furthermore we demonstrate that PSK, when given in combination with paclitaxel can enhance tumor regression. We hypothesize that the addition of PSK to standard therapy with paclitaxel and trastuzumab will augment anti-tumor immunity and result in improved PCR rate and overall survival in neu transgenic mice bearing spontaneous tumors.
The Specific Aims of the proposal are to: (1) Determine whether the addition of PSK to standard neoadjuvant therapy for HER2+/ER- LABC will increase the rate of PCR and overall survival in neu-transgenic mice, (2) Determine whether the addition of PSK to standard neoadjuvant therapy for HER2+/ER- LABC results in the generation of a pro-inflammatory tumor microenvironment that supports anti-tumor immunity, and (3) Determine the potential augmentation of a systemic (adaptive) immune response elicited by incorporating PSK into standard neoadjuvant therapy for HER2+/ER- LABC. Data generated here will lay the foundation for the potential integration of complementary and alternative medicine (CAM) therapy into the neoadjuvant treatment of LABC.

Characterization Of Potential Antigens For Immunization Against Colon Cancer In Multiple Rodent Colon Tumor Models And Testing Multivalent Tumor Vaccines in Min Mice

Grant Number: HHSN261-200533000C

The successful identification of novel tumor antigens is critical to cancer immunotherapy and its potential use in cancer prevention. Colon cancer is the third most common cancer in both men and women, and is the second most common cause of cancer death. There are a variety of approaches towards deciding upon antigens which might be employed to develop a colon cancer vaccine. In the rare cases where there is a highly over expressed protein that appears to be directly associated with tumor development, this appears to be a promising approach both in animal models and clinically. A second is to pick antigens that are overexpressed in tumors and that appear to be associated with the tumor process. More recently, there has been an attempt to determine whether within the group of over expressed antigens there may be some that have elicited either an antibody response or a T cell mediated response in individuals with the tumor. The particular appeal of these antigens is that individuals have responded to these proteins and by implication it may be easier to break tolerance against these specific antigens.
The objective of this work assignment would be to:
(1) Based on expression data determine whether expression of specific proteins, as determined by RNA when comparing colon lesions and normal mucosa, is similar in human colon tumors and certain animal models (AOM mouse, Min mouse and AOM rat).
(2) Determine whether animals bearing colon or intestinal lesions demonstrate immunologic responses as determined by SEREX or specific T cell reactivity against relevant antigens and compare this with relevant proteins which have elicited an immune response in persons bearing colon tumors.
(3) From these proteins develop four antigens which appear amenable to development as potential antigens in a multivalent vaccine.
(4) Test these antigens for their ability to elicit an immune response in at least two strains of mice or in one strain of mice and in rats.
(5) Test the potential vaccine (4 antigens vs 2 antigens and 2 antigens) to inhibit tumor formation in Min mice on an F1 background which may decrease the number of intestinal lesions. The advantage of the F1 mice is that the reduced number of lesions may offer additional time in which to examine the efficacy of the proposed vaccine.
(6) Test the vaccine together with a sub-ideal dose of a chemopreventive agent e.g. Naproxen

Breast Cancer Integrative Oncology: Prospective Matched Controlled Outcomes Study

Grant Number: H33B11-03(4) prime (5R01AT005873-04)

Purpose:
To conduct a prospective outcomes study describing use of Complementary and Alternative medicine (CAM), and integrated oncology (IO) and their effects on breast cancer patients in community settings.

Background:
Between 50% and 80% of women in the United States with breast cancer supplement their conventional medical treatment regimen with some form of CAM therapy. Most CAM use is self-prescribed and unsupervised, however, IO has matured into a set of science-based practices that warrant evaluation. IO clinics are directed by physicians, MDs, naturopathic physicians (NDs), and doctors of acupuncture and Oriental medicine (DAOM), who use best practices and evidence-based CAM to improve the health and well being of cancer patients who also receive standard conventional cancer therapy. This study will describe IO care as provided in community clinics and provide an initial evaluation of the effects of IO care and CAM use on breast cancer patients’ Health Related Quality of Life (HRQOL).

Hypothesis:
IO services improve patients’ quality of life and decrease cancer recurrence rates in breast cancer patients, as compared to women with similar disease states and prognoses who do not receive IO care, and may or may not use CAM treatment on their own.

Methods:
A partnership between Bastyr University and Fred Hutchinson Cancer Research Center will create a case-control matched cohort of IO using breast cancer patients and breast cancer patients with similar prognosis at time of diagnosis who did not use IO care and may or may not have used CAM on a self-prescription basis. This cohort will be followed prospectively allowing for a description of the outcomes of IO breast cancer care provided through a community IO clinic network in Seattle.
Aims: 1) To describe the treatments received by breast cancer patients receiving physician level integrative oncology (IO) care from community clinics (cohort; n = 600) and the cost of IO care for breast cancer; 2) To describe the Health Related Quality of Life (HRQOL) experienced by breast cancer patients receiving physician level Integrative Oncology (IO) care (cohort; n = 300) and that of a group of comparison women with breast cancer (cohort; n = 900) receiving conventional treatment with or without self-prescribed CAM (n = 400-450 and n = 450-500). In addition to describing changes in HRQOL associated with IO and CAM use and estimating the effects attributable to IO or CAM use, 3) To collect pilot data on survival and disease free survival in outcomes at 2 and 3 years follow-up in a cohort of 300 IO breast cancer patients, and a matched comparison group of 900 who did not choose to use IO during their initial treatment for breast cancer.

Pages