Evaluation of BH3 mimetics as a combination therapy with irradiation in head and neck squamous cell carcinoma

Biomed Pharmacother. 2024 Jun:175:116719. doi: 10.1016/j.biopha.2024.116719. Epub 2024 May 15.

Abstract

Introduction: Head and neck squamous cell carcinoma (HNSCC) is a common cancer with a five-year survival rate around 60%, indicating a need for new treatments. BH3 mimetics are small molecules that inhibit anti-apoptotic Bcl-2 family proteins, resulting in apoptosis induction.

Methods: We performed a high-throughput screen using a Myogel matrix to identify the synergy between irradiation and the novel BH3 mimetics A-1155463, A-1331852, and navitoclax in 12 HNSCC cell lines, normal (NOF) and cancer-associated fibroblasts (CAF), and dysplastic keratinocytes (ODA). Next, we examined synergy in an apoptosis assay, followed by a clonogenic assay and a Myogel spheroid on selected HNSCC cell lines. Finally, we applied zebrafish larvae xenograft to validate the effects of navitoclax and A-1331852.

Results: All three BH3 mimetics exhibited a strong synergy with irradiation in eight HNSCC cell lines and ODAs, but not in NOFs and CAFs. A-1155463 and A-1331852 induced apoptosis and reduced proliferation, and together with irradiation, significantly increased apoptosis and arrested proliferation. A-1331852 and navitoclax significantly decreased the clonogenicity compared with the control, and combination treatment led to a decreased clonogenicity compared with monotherapy or irradiation. However, unlike navitoclax or A-1155463, only A-1331852 significantly reduced cancer cell invasion. Furthermore, in spheroid and zebrafish, irradiation appeared ineffective and failed to significantly increase the drug effect. In the zebrafish, A-1331852 and navitoclax significantly reduced the tumor area and metastasis.

Conclusions: Our findings encourage the further preclinical investigation of BH3 mimetics, particularly A-1331852, as a single agent or combined with irradiation as a treatment for HNSCC.

Keywords: A-1155463; A-1331852; Apoptosis assay; BH3 mimetics; Clonogenic survival assay; Combination therapy; Head and neck squamous cell carcinoma; High-throughput drug combination screen; Invasion assay; Navitoclax; Xenograft.

MeSH terms

  • Aniline Compounds / pharmacology
  • Animals
  • Antineoplastic Agents / pharmacology
  • Apoptosis* / drug effects
  • Carcinoma, Squamous Cell / drug therapy
  • Carcinoma, Squamous Cell / pathology
  • Carcinoma, Squamous Cell / radiotherapy
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Combined Modality Therapy
  • Head and Neck Neoplasms* / drug therapy
  • Head and Neck Neoplasms* / pathology
  • Head and Neck Neoplasms* / radiotherapy
  • Humans
  • Peptide Fragments
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Squamous Cell Carcinoma of Head and Neck* / drug therapy
  • Squamous Cell Carcinoma of Head and Neck* / pathology
  • Squamous Cell Carcinoma of Head and Neck* / radiotherapy
  • Sulfonamides / pharmacology
  • Xenograft Model Antitumor Assays
  • Zebrafish*

Substances

  • navitoclax
  • Aniline Compounds
  • Sulfonamides
  • Bax protein (53-86)
  • Antineoplastic Agents
  • Proto-Oncogene Proteins c-bcl-2
  • Peptide Fragments
  • Proto-Oncogene Proteins