High 4-1BB Expression in PBMCs and Tumor Infiltrating Lymphocytes (TILs) in Patients with Head and Neck Squamous Cell Carcinoma

Abstract Objective  4-1BB is a costimulatory immune-activating molecule. Increased amounts of this protein have previously been found in the plasma of patients with oropharyngeal and oral cancer. Here, we focused on this molecule that functions as part of the immune system. We investigated 4-1BB in the peripheral blood mononuclear cells (PBMCs) and tumor infiltrating lymphocytes (TILs) of patients with head and neck squamous cell cancer (HNSCC). Materials and Methods  The expression level of 4-1BB in the PBMCs was determined using real-time polymerase chain reaction (PCR). The TIMER (Tumor Immune Estimation Resource) web server was utilized to approximate the 4-1BB level in HNSCC TILs. Moreover, 4-1BB immunohistochemistry (IHC) was used to validate TILs in four organs of HNSCC, including oral cancer (OC), oropharyngeal cancer (OPC), sinonasal cancer (SNC), and laryngeal cancer (LC), in both the tumor area and adjacent normal epithelium. The difference in 4-1BB expression levels in various groups was assessed using a Kruskal-Wallis test and an independent sample t-test. Results  The level of 4-1BB expression in PBMCs was highest in OPC, followed by OC and healthy controls (HC). Significant differences were discovered between HC and OPC and between OC and OPC. Bioinformatics revealed a substantial correlation between 4-1BB expression level and lymphocyte infiltration in HNSCC, including B cells, CD8+ T cells, and CD4+ T cells. IHC validation in HNSCC tissue revealed that the average number of 4-1BB positive TILs in all four HNSCC subtypes was considerably greater than the number of lymphocytes seen in adjacent normal tissue. Interestingly, the number of lymphocytes that were 4-1BB positive increased in relation to the TIL level. Conclusion  A higher number of 4-1BB expression levels were found in the PBMCs and TILs of HNSCC patients, implying that 4-1BB may be a promising approach for HNSCC patients to improve their immune function. It is important to study and create a treatment that uses 4-1BB medicine as well as existing drugs.

[1]  Songzhe Zhu,et al.  Tumor microenvironment and immunotherapy of oral cancer , 2022, European Journal of Medical Research.

[2]  N. Rizvi,et al.  First-in-human study of an OX40 (ivuxolimab) and 4-1BB (utomilumab) agonistic antibody combination in patients with advanced solid tumors , 2022, Journal for ImmunoTherapy of Cancer.

[3]  Yu Ding,et al.  Development and characterization of a novel human CD137 agonistic antibody with anti‐tumor activity and a good safety profile in non‐human primates , 2022, FEBS open bio.

[4]  Seung-Oe Lim,et al.  4-1BB: A promising target for cancer immunotherapy , 2022, Frontiers in Oncology.

[5]  N. Kitkumthorn,et al.  Immune-associated plasma proteins in oral and oropharyngeal cancer patients , 2022, Heliyon.

[6]  Craig B. Davis,et al.  Utomilumab in Patients With Immune Checkpoint Inhibitor-Refractory Melanoma and Non-Small-Cell Lung Cancer , 2022, Frontiers in Immunology.

[7]  L. Pacini,et al.  EGFR Exon 20 Insertion Mutations in Sinonasal Squamous Cell Carcinoma , 2022, Cancers.

[8]  E. Marchioni,et al.  Complete Response to Nivolumab in Recurrent/Metastatic HPV-Positive Head and Neck Squamous Cell Carcinoma Patient After Progressive Multifocal Leukoencephalopathy: A Case Report , 2022, Frontiers in Oncology.

[9]  A. Jemal,et al.  Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries , 2021, CA: a cancer journal for clinicians.

[10]  C. R. Leemans,et al.  Head and neck squamous cell carcinoma , 2020, Nature Reviews Disease Primers.

[11]  Xiaole Shirley Liu,et al.  TIMER2.0 for analysis of tumor-infiltrating immune cells , 2020, Nucleic Acids Res..

[12]  T. Tsuzuki,et al.  Immune‐checkpoint molecules on regulatory T‐cells as a potential therapeutic target in head and neck squamous cell cancers , 2020, Cancer science.

[13]  Craig B. Davis,et al.  A phase Ib study of utomilumab (PF-05082566) in combination with mogamulizumab in patients with advanced solid tumors , 2019, Journal of Immunotherapy for Cancer.

[14]  C. Glorieux,et al.  Regulation of CD137 expression through K-Ras signaling in pancreatic cancer cells , 2019, Cancer communications.

[15]  F. Warburton,et al.  Clinical evaluation of tumour‐infiltrating lymphocytes as a prognostic factor in patients with human papillomavirus‐associated oropharyngeal squamous cell carcinoma , 2019, Histopathology.

[16]  Vo Truong Nhu Ngoc,et al.  An Update on Anti-CD137 Antibodies in Immunotherapies for Cancer , 2019, International journal of molecular sciences.

[17]  Craig B. Davis,et al.  Phase I Study of Single-Agent Utomilumab (PF-05082566), a 4-1BB/CD137 Agonist, in Patients with Advanced Cancer , 2018, Clinical Cancer Research.

[18]  I. Melero,et al.  Immunotherapy targeting 4-1BB: mechanistic rationale, clinical results, and future strategies. , 2018, Blood.

[19]  M. Roach,et al.  Handbook of Evidence-Based Radiation Oncology , 2018, Springer International Publishing.

[20]  J. Doescher,et al.  [The 8th edition of the AJCC Cancer Staging Manual : Updates in otorhinolaryngology, head and neck surgery]. , 2017, HNO.

[21]  Jun S. Liu,et al.  TIMER: A Web Server for Comprehensive Analysis of Tumor-Infiltrating Immune Cells. , 2017, Cancer research.

[22]  H. Kohrt,et al.  4-1BB agonism: adding the accelerator to cancer immunotherapy , 2016, Cancer Immunology, Immunotherapy.

[23]  S. Subramanian,et al.  Oral Cancer: Prevention, Early Detection, and Treatment , 2015 .

[24]  M. Oka,et al.  Combination immunotherapy with 4-1BB activation and PD-1 blockade enhances antitumor efficacy in a mouse model of subcutaneous tumor. , 2015, Anticancer research.

[25]  M. Hermsen,et al.  Sinonasal carcinoma: clinical, pathological, genetic and therapeutic advances , 2014, Nature Reviews Clinical Oncology.

[26]  J. Allison,et al.  Combination CTLA-4 Blockade and 4-1BB Activation Enhances Tumor Rejection by Increasing T-Cell Infiltration, Proliferation, and Cytokine Production , 2011, PloS one.

[27]  B. Lu,et al.  Characterization and application of three novel monoclonal antibodies against human 4-1BB: distinct epitopes of human 4-1BB on lung tumor cells and immune cells. , 2007, Tissue antigens.

[28]  Sergio L. Schmukler,et al.  Emerging Market Instability : Do Sovereign Ratings Affect Country Risk and Stock Returns ? , 1997 .

[29]  R. Hermans.  Oropharyngeal cancer , 2005, Cancer imaging : the official publication of the International Cancer Imaging Society.

[30]  G. Mufti,et al.  Role of 4-1BB:4-1BB ligand in cancer immunotherapy , 2004, Cancer Gene Therapy.

[31]  T. Watts,et al.  4-1BB Ligand-Mediated Costimulation of Human T Cells Induces CD4 and CD8 T Cell Expansion, Cytokine Production, and the Development of Cytolytic Effector Function1 , 2002, The Journal of Immunology.