Induction of Invasive Transitional Cell Bladder Carcinoma in Immune Intact Human MUC1 Transgenic Mice:  A Model for Immunotherapy Development

Daniel P. Vang; Gregory T. Wurz; Stephen M. Griffey; Chiao-Jung Kao; Audrey M. Gutierrez; Gregory K. Hanson; Michael Wolf; Michael W. DeGregorio

doi:10.3791/50868

A subscription to JoVE is required to view this content. Sign in or start your free trial.

Method Article

Induction of Invasive Transitional Cell Bladder Carcinoma in Immune Intact Human MUC1 Transgenic Mice: A Model for Immunotherapy Development

21.1K views

⸱

DOI:

10.3791/50868

⸱

October 30th, 2013

Daniel P. Vang¹ , Gregory T. Wurz¹ , Stephen M. Griffey² , Chiao-Jung Kao¹ , Audrey M. Gutierrez¹ , Gregory K. Hanson¹ , Michael Wolf³ , Michael W. DeGregorio¹

¹Department of Internal Medicine, Division of Hematology and Oncology, University of California, Davis, ²Comparative Pathology Laboratory, UC Davis School of Veterinary Medicine, University of California, Davis, ³Merck Serono Research, Merck KGaA, Darmstadt, Germany

Summary

An N-butyl-N-(4-hydroxybutyl)nitrosamine-induced bladder cancer model was developed in human mucin 1 (MUC1) transgenic mice for the purpose of testing MUC1-directed immunotherapy. After administering a MUC1-targeted peptide vaccine, a cytotoxic T lymphocyte response to MUC1 was confirmed by measuring serum cytokine levels and T-cell specific activity.

Abstract

A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy and/or cytotoxic chemotherapy. To induce bladder cancer, C57BL/6 mice (MUC1.Tg and wild type) were treated orally with the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN) at 3.0 mg/day, 5 days/week for 12 weeks. To assess the effects of OH-BBN on serum cytokine profile during tumor development, whole blood was collected via submandibular bleeds prior to treatment and every four weeks. In addition, a MUC1-targeted peptide vaccine and placebo were administered to groups of mice weekly for eight weeks. Multiplex fluorometric microbead immunoanalyses of serum cytokines during tumor development and following vaccination were performed. At termination, interferon gamma (IFN-γ)/interleukin-4 (IL-4) ELISpot analysis for MUC1 specific T-cell immune response and histopathological evaluations of tumor type and grade were performed. The results showed that: (1) the incidence of bladder cancer in both MUC1.Tg and wild type mice was 67%; (2) transitional cell carcinomas (TCC) developed at a 2:1 ratio compared to squamous cell carcinomas (SCC); (3) inflammatory cytokines increased with time during tumor development; and (4) administration of the peptide vaccine induces a Th1-polarized serum cytokine profile and a MUC1 specific T-cell response. All tumors in MUC1.Tg mice were positive for MUC1 expression, and half of all tumors in MUC1.Tg and wild type mice were invasive. In conclusion, using a team approach through the coordination of the efforts of pharmacologists, immunologists, pathologists and molecular biologists, we have developed an immune intact transgenic mouse model of bladder cancer that expresses hMUC1.

Introduction

Bladder cancer is the fourth most common form of cancer and the eighth leading cause of cancer deaths in American men. In the United States, an estimated 72,500 new cases and 15,000 deaths from bladder cancer are expected among men and women combined in 2013¹. The incidence of bladder cancer is approximately three times as high in men compared to women. In the United States, transitional cell carcinomas (TCC) account for over 90% of cases, while squamous cell carcinomas (SCC) have an incidence of less than 2%². The overall relative 5-year survival rate for papillary TCC is 91.5% compared to only 30.9% for SCC². Although noninvasive pap....