Intravesical Immunomodulatory Imiquimod Enhances Bacillus Calmette-Guérin Downregulation of Nonmuscle-invasive Bladder Cancer

Clin Genitourin Cancer. 2018 Jun;16(3):e587-e593. doi: 10.1016/j.clgc.2017.10.019. Epub 2017 Nov 7.

Abstract

Background: The Toll-like receptor (TLR)2/4 agonist bacillus Calmette-Guérin (BCG), although not failure proof, has been the most efficient immunomodulatory treatment of immunogenic nonmuscle-invasive bladder cancer (NMIBC) for > 40 years. We investigated the role of the immunomodulatory molecule TLR7 agonist imiquimod through the BCG key receptors TLR2/4 and the main downstream molecules of the mammalian target of rapamycin pathway in NMIBC treatment.

Materials and methods: A total of 40 Fischer-344 rats, 7 weeks old, received 4 doses of 1.5 mg/kg N-methyl-N-nitrosourea intravesically on weeks 0, 2, 4, and 6 for cancer induction. At week 8, the rats were randomized into 4 groups (10 per group) and treated intravesically once a week for 6 weeks: control (0.2 mL of vehicle); BCG (2 × 106 colony-forming units Connaught strain in 0.2 mL); imiquimod (20 mg/kg in 0.2 mL), and associated treatment BCG plus imiquimod in 0.2 mL. The bladders were extracted and analyzed for histopathology, immunohistochemistry, cell proliferation (Ki-67), apoptosis (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling [TUNEL]), and immunoblotting for TLR2, TLR4, p-P70S6K, and p-4E-BP1 proteins.

Results: The histopathology results showed that BCG and imiquimod decreased bladder tumorigenesis compared with the control group, with a proliferation decrease (Ki-67) and an apoptosis increase (TUNEL). BCG upregulated TLR2/4, imiquimod upregulated TLR4, and both downregulated P70S6K1.

Conclusion: Imiquimod is able to efficiently decrease bladder carcinogenesis through upregulation of TLR7/4 and downregulation of P70S6K1 protein, generating new perspectives to boost BCG effects in the future.

Keywords: BCG; Intravesical; Toll-like; Urothelial cancer; mTOR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / administration & dosage*
  • Adjuvants, Immunologic / pharmacology
  • Administration, Intravesical
  • Animals
  • BCG Vaccine / administration & dosage*
  • BCG Vaccine / pharmacology
  • Cell Proliferation / drug effects
  • Drug Synergism
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Imiquimod / administration & dosage*
  • Imiquimod / pharmacology
  • Methylnitrosourea / adverse effects*
  • Random Allocation
  • Rats
  • Rats, Inbred F344
  • Ribosomal Protein S6 Kinases, 70-kDa / metabolism
  • Toll-Like Receptor 2 / metabolism
  • Toll-Like Receptor 4 / metabolism
  • Treatment Outcome
  • Urinary Bladder Neoplasms / chemically induced
  • Urinary Bladder Neoplasms / drug therapy*
  • Xenograft Model Antitumor Assays

Substances

  • Adjuvants, Immunologic
  • BCG Vaccine
  • Tlr2 protein, rat
  • Tlr4 protein, rat
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Methylnitrosourea
  • Ribosomal Protein S6 Kinases, 70-kDa
  • Imiquimod