Influence of tumor-associated macrophages and HLA class I expression according to HPV status in head and neck cancer patients receiving chemo/bioradiotherapy

doi:10.1016/j.radonc.2018.08.013

Radiotherapy and Oncology

Volume 130, January 2019, Pages 89-96

https://doi.org/10.1016/j.radonc.2018.08.013 Get rights and content

Highlights

•
Intraepithelial macrophage density was associated with favorable progression-free survival.
•
HLA class I down-regulation was not an independent prognostic factor.
•
Stromal M2 density may potentially be used to guide treatment selection.

Abstract

Background and purpose

To investigate the prognostic value of tumor-associated macrophages (TAM) and HLA class I expression according to HPV status in patients with head and neck squamous cell carcinoma treated with definitive radiotherapy combining cisplatin (CRT) or cetuximab (BRT).

Material and methods

Ninety-five patients were enrolled. The density of CD68+ cells and CD68+ CD163+ cells (further referred as M2) in the intraepithelial and the stromal compartments, respectively, as well as HLA class I expression in tumor cells, were evaluated semi-quantitatively. Correlations between biomarker expression and treatment outcomes were analyzed.

Results

Multivariate analysis showed that the intraepithelial macrophage density (IEMD) was prognostic for favorable progression-free survival (PFS) and there was a non-significant trend for improved overall survival (OS). HLA class I down-regulation was not an independent prognostic factor. Subgroup analysis showed that in p16+ population, patients with high IEMD had improved 5-year PFS vs. patients with low IEMD (81.2% vs. 25.0%, p < 0.001), while in p16− population, no difference was observed. Similarly, when stratified by primary tumor site, IEMD showed prognostic value in oropharyngeal cancer patients (OPC) but not non-OPC patients. Five-year PFS of patients with low stromal M2 macrophage density treated with CRT was significantly improved vs. those with BRT (54.5% vs. 36.1%, p = 0.03), while in tumors with high M2, there was no significant difference (50.3% vs. 42.9%, p = 0.67).

Conclusions

The prognostic role of TAM phenotype and distribution depends on HPV status and might predict treatment response. They prompt further validation in prospective studies.

Section snippets

Patients and tissues

Patients were selected from the former study cohort of 265 patients with histologically confirmed HNSCC, diagnosed between 2006 and 2012 in our institute (CRT: n = 194, BRT: n = 71) [17], [18]. Patients with stage III–IVb disease according to American Joint Committee on Cancer (AJCC)/International Union for Cancer Control (UICC) TNM classification 2010, received total radiation dose of 70 Gy, ≥2 cycles of concurrent cisplatin or ≥3 cycles of concurrent cetuximab were selected from the initial

Patient characteristics

The clinical and pathological characteristics of the patients included in this study are summarized in Table 1. The age of the study population ranged from 38 to 81 years (median, 60 years). Most (80%) patients were male. Sixty-four (67%) patients were never/former smokers and 31 (33%) were current smokers. With regard to HPV status, 27 (28%) patients had p16+ tumors and 68 (72%) had p16− tumors. All patients with p16+ tumors were oropharyngeal cancer (OPC) patients. Information on p16 status

Discussion

Tumor microenvironment consists of various host components including stromal cells, blood vessels and inflammatory infiltrates. It plays an important role in carcinogenesis, tumor progression and response to therapeutic interventions [21]. In the present study, although macrophage density and HLA class I expression were not identified as independent prognostic factors by multivariate analysis, subgroup analysis showed that intraepithelial macrophage density may play different roles in HPV+ as

Conflict of interest statement

None declared.

Acknowledgements

This work was supported by EU 7th framework program (ARTFORCE), No grant number is applicable; and China Scholarship Council (CSC) [201406105021 to D. O.].

References (43)

J.P. Pignon et al.
Meta-analysis of chemotherapy in head and neck cancer (MACH-NC): an update on 93 randomised trials and 17,346 patients
Radiother Oncol
(2009)
H. Mirghani et al.
Increased radiosensitivity of HPV-positive head and neck cancers: Molecular basis and therapeutic perspectives
Cancer Treat Rev
(2015)
A. Levy et al.
Radiation therapy and immunotherapy: implications for a combined cancer treatment
Crit Rev Oncol Hematol
(2013)
M. Weber et al.
Prognostic significance of macrophage polarization in early stage oral squamous cell carcinomas
Oral Oncol
(2016)
F. Concha-Benavente et al.
Immunological and clinical significance of HLA class I antigen processing machinery component defects in malignant cells
Oral Oncol
(2016)
D. Ou et al.
Concurrent chemoradiotherapy with cisplatin or cetuximab for locally advanced head and neck squamous cell carcinomas: does human papilloma virus play a role?
Oral Oncol
(2016)
N.F. Schlecht et al.
A comparison of clinically utilized human papillomavirus detection methods in head and neck cancer
Mod Pathol
(2011)
M.R. Galdiero et al.
Tumor associated macrophages and neutrophils in cancer
Immunobiology
(2013)
B. Ruffell et al.
Macrophages and therapeutic resistance in cancer
Cancer Cell
(2015)
Y.S. Lee et al.
Composition of inflammatory cells regulating the response to concurrent chemoradiation therapy for HPV (+) tonsil cancer
Oral Oncol
(2015)

P.J. Murray et al.

Macrophage activation and polarization: nomenclature and experimental guidelines

Immunity

(2014)

L. Galluzzi et al.

Immunological effects of conventional chemotherapy and targeted anticancer agents

Cancer Cell

(2015)

J. Li et al.

Cetuximab ameliorates suppressive phenotypes of myeloid antigen presenting cells in head and neck cancer patients

J Immunother Cancer

(2015)

Y. Tao et al.

Advances in radiotherapy of head and neck cancers

Curr Opin Oncol

(2010)

L.Q. Chow et al.

Antitumor activity of pembrolizumab in biomarker-unselected patients with recurrent and/or metastatic head and neck squamous cell carcinoma: results from the phase Ib KEYNOTE-012 expansion cohort

J Clin Oncol

(2016)

K.K. Ang et al.

Human papillomavirus and survival of patients with oropharyngeal cancer

N Engl J Med

(2010)

P. Italiani et al.

From monocytes to M1/M2 macrophages: phenotypical vs functional differentiation

Front Immunol

(2014)

C. Li et al.

Infiltration of tumor-associated macrophages in human oral squamous cell carcinoma

Oncol Rep

(2002)

S.K. Lau et al.

CD163: a specific marker of macrophages in paraffin-embedded tissue samples

Am J Clin Pathol

(2004)

N. Fujii et al.

Cancer-associated fibroblasts and CD163-positive macrophages in oral squamous cell carcinoma: their clinicopathological and prognostic significance

J Oral Pathol Med

(2012)

B. Seliger

Molecular mechanisms of HLA class I-mediated immune evasion of human tumors and their role in resistance to immunotherapies: HLA class I-mediated immune escape in tumors

HLA

(2016)

Cited by (24)

Tumor associated macrophage in HPV<sup>+</sup> tumors: Between immunosuppression and inflammation
2023, Seminars in Immunology
Citation Excerpt :
Indeed, in a multivariate analysis, the authors showed that HPV-16 was required to increase intraepithelial macrophage density and HLA-class I expression, which leads to a better 5-year progression free survival in oropharyngeal patients (Table 1). However, the macrophage phenotype was not described in this study and therefore avoid any comparison with studies showing poorer prognosis in TAM infiltrated HNSCC [81]. Interestingly, HPV strains are able to favor certain types of TAM to rise and participate to the TME.
Over the past few decades, with the rise of immunotherapies, tumor infiltrating immune cells were increasingly investigated. Indeed, they may represent biomarkers for patient outcome prediction, they may bear immune checkpoint markers that can be targeted by therapeutic antibodies and mechanistic studies may reveal how to tweak their activation profile so that we can re-direct them towards tumor cells. Macrophages possess a central place in tissue homeostasis for tissue remodeling and cleaning, transformed cell elimination, phagocytosis and regulation of inflammation via cytokine production. All these functions allow the discovery of approaches to target Tumor Associated Macrophages (TAMs) using immunotherapies. Indeed, TAMs express known immune checkpoint markers such as PD-L1, CD40, Sirp-α and markers such as CD163, CD204, TREM2, TREM1 associated with prognosis. In the context of therapies TAM may participate to antibody dependent cell phagocytosis (ADCP) thanks to FCγ−Receptors. Here, we will review the recent literature on TAMs in the specific context of HPV⁺ tumors. Indeed, HPV infection of mucosal tissue may lead to head and neck, cervical, penile, anal and vaginal cancers. HPV⁺ tumors exhibit a higher immune cell infiltrate, which relies on inflammation, immunosuppression and anti-viral response. In this context, and considering the many functions on macrophages, we will show the versatility of TAMs in a tumor microenvironment with viral infection features.
The prognostic role of tumor associated macrophages in squamous cell carcinoma of the head and neck: A systematic review and meta-analysis
2022, Oral Oncology
Citation Excerpt :
Pooled meta-analysis of the final ten studies investigating the relationship between CD68+TAMs and OS showed that a low number of CD68+TAMs correlated to a better OS (HR 1.36 95 %CI (1.07–1.72) P = .01, Figure 2b). In the univariate and multivariate pooled analysis, a total of four studies assessed CD68 in both tumor epithelium and tumor stroma.[21,25,37,43] Kikuchi et al. (2020) reported a trend that low CD68+TAMs correlated to better OS in the tumor epithelium.
Head and neck squamous cell carcinoma (HNSCC) is an immunogenic cancer type, and tumor associated macrophages (TAMs) are a major component of the tumor microenvironment (TME). In this systematic review and meta-analysis, studies assessing tumor infiltration with CD68+, iNOS+, HLA-DR+, CD11b+, CD163+, CD206+, and CD204+TAMs were included, and correlation to survival hazard was studied. A low number of CD68+TAMs correlated to better overall survival (OS) in multivariate analysis (HR 1.36 95 %CI (1.07–1.72) P = .01). CD68+TAMs did not correlate to disease free survival (DFS), disease specific survival (DSS), progression free survival (PFS), or recurrence free survival (RFS). A low number of CD163+TAMs correlated to better OS in uni- and multivariate analysis (resp. HR 2.65 95 %CI (1.57–4.46) P = .01 and HR 2.42 95 %CI (1.72–3.41) P < .001). A low number of CD163+TAMs also correlated to better DFS and PFS, whereas a low number of CD204+TAMs only correlated to PFS. While IHC analysis of pan macrophage marker CD68 and M2-like marker CD163 both show prognostic utility in OS, CD163 is a stronger prognosticator, as indicated by multivariate meta-analysis. CD163+TAMs also correlate to DFS and PFS; outcomes that are more relevant to patients, thus showing promising results for future clinical implementation.
Precision medicine and immuno-radiotherapy
2021, Cancer/Radiotherapie
De nombreuses études cliniques visent à intégrer l’immunothérapie en oncologie radiothérapie, soit pour favoriser des réponses abscopales en association à la radiothérapie chez des patients atteints de cancer métastatique, soit dans le cadre du traitement d’une tumeur localement évoluée. La recherche de biomarqueurs de réponse au traitement est un axe majeur du développement de ces associations thérapeutiques, pour permettre l’identification précoce des patients qui tireront bénéfice du traitement, dans une approche personnalisée. Nous passons en revue certaines des stratégies applicables pour la personnalisation aux traitements associant radiothérapie et immunothérapie.
Numerous clinical studies aim to integrate immunotherapy in radiotherapy oncology, either for generating abscopal responses in metastatic patients in combination with radiotherapy, or in the treatment of a locally advanced tumor. The search for biomarkers of response to treatment is a major axis in the development of these therapeutic combinations, to allow the early identification of patients who will benefit from the treatment, in the context of an increasingly personalized approach. We review some of the strategies that can be applied for personalization to combined radiotherapy and immunotherapy treatments.
Correlation between HPV infection and ovarian epithelial cancer diagnosed by Dynamic Contrast-enhanced magnetic resonance imaging information technology under exponential distribution mathematical model
2021, Results in Physics
This research aims to analyze the correlation between human papillomavirus (HPV) infection and ovarian epithelial cancer based on exponential distribution mathematical model, so as to provide an experimental basis for the early diagnosis of ovarian epithelial cancer by magnetic resonance imaging (MRI) in the future. In this study, 124 patients with ovarian epithelial cancer tissues pathologically confirmed in our hospital from March 31, 2017 to February 20, 2019 were selected as the experimental group, and 64 patients with normal ovarian tissues were selected as the control group, and the positive rate of HPV infection was detected by the computer cell test (CCT) system. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was constructed and adopted to analyze the scanning images. The correlation between HPV infection and pathological type, clinical stage, tissue differentiation degree, and CA125 in serum was studied by exponential distribution mathematical model. The results showed that the positive rate of HPV infection in the ovarian tissues of the experimental group was significantly higher than that of the control group (P < 0.05). Highly differentiated patients accounted for 25.97%, moderately differentiated patients accounted for 43.81%, and lowly differentiated patients accounted for 30.22%. Patients with clinical stage I-II accounted for 24.72%, patients with stage III accounted for 49.11%, and patients with stage IV accounted for 26.17%. HPV infection was significantly correlated with clinical stage III and moderate tissue differentiation by MRI of patients (P < 0.05), and extremely significantly correlated with clinical stage IV and low tissue differentiation by MRI (P < 0.001). The above findings show that the information technology of dynamic contrast-enhanced magnetic resonance imaging can clearly display the lesion metastasis of ovarian epithelial cancer patients with high soft tissue resolution.
High throughput profiling of undifferentiated pleomorphic sarcomas identifies two main subgroups with distinct immune profile, clinical outcome and sensitivity to targeted therapies
2020, EBioMedicine
Citation Excerpt :
Amplification and detection steps were performed with an Ultraview kit and 3,3′-diaminobenzidine was used as a chromogen. CD68/CD163 double staining was performed on a Ventana Discovery Ultra platform (Roche, Bâle, Switzerland) and image analysis as described previously [13]. Image analysis was performed under the supervision of a pathologist (JA) to detect the density of CD8 and PD-1 positive cells in the tumor areas and the proportion of IDO1, CD68 and CD163 stained surface of tumor slides or spots, as previously described [14].
Undifferentiated pleomorphic sarcoma (UPS) is the most frequent, aggressive and less-characterized sarcoma subtype. This study aims to assess UPS molecular characteristics and identify specific therapeutic targets.
High-throughput technologies encompassing immunohistochemistry, RNA-sequencing, whole exome-sequencing, mass spectrometry, as well as radiomics were used to characterize three independent cohorts of 110, 25 and 41 UPS selected after histological review performed by an expert pathologist. Correlations were made with clinical outcome. Cell lines and xenografts were derived from human samples for functional experiments.
CD8 positive cell density was independently associated with metastatic behavior and prognosis. RNA-sequencing identified two main groups: the group A, enriched in genes involved in development and stemness, including FGFR2, and the group B, strongly enriched in genes involved in immunity. Immune infiltrate patterns on tumor samples were highly predictive of gene expression classification, leading to call the group B ‘immune-high’ and the group A ‘immune-low’. This molecular classification and its prognostic impact were confirmed on an independent cohort of UPS from TCGA. Copy numbers alterations were significantly more frequent in immune-low UPS. Proteomic analysis identified two main proteomic groups that highly correlated with the two main transcriptomic groups. A set of nine radiomic features from conventional MRI sequences provided the basis for a radiomics signature that could select immune-high UPS on their pre-therapeutic imaging. Finally, in vitro and in vivo anti-tumor activity of FGFR inhibitor JNJ-42756493 was selectively shown in cell lines and patient-derived xenograft models derived from immune-low UPS.
Two main disease entities of UPS, with distinct immune phenotypes, prognosis, molecular features and MRI textures, as well as differential sensitivity to specific anticancer agents were identified. Immune-high UPS may be the best candidates for immune checkpoint inhibitors, whereas this study provides rational for assessing FGFR inhibition in immune-low UPS.
This work was partly founded by a grant from La Ligue.
Prognostic Significance of the Microenvironment in Human Papillomavirus Oropharyngeal Carcinoma: A Systematic Review
2024, Laryngoscope

View all citing articles on Scopus

View full text

Original articleInfluence of tumor-associated macrophages and HLA class I expression according to HPV status in head and neck cancer patients receiving chemo/bioradiotherapy

Highlights

Abstract

Background and purpose

Material and methods

Results

Conclusions

Section snippets

Patients and tissues

Patient characteristics

Discussion

Conflict of interest statement

Acknowledgements

Radiother Oncol

Cancer Treat Rev

Crit Rev Oncol Hematol

Oral Oncol

Oral Oncol

Oral Oncol

Mod Pathol

Immunobiology

Cancer Cell

Oral Oncol

Immunity

Cancer Cell

J Immunother Cancer

Advances in radiotherapy of head and neck cancers

Curr Opin Oncol

Antitumor activity of pembrolizumab in biomarker-unselected patients with recurrent and/or metastatic head and neck squamous cell carcinoma: results from the phase Ib KEYNOTE-012 expansion cohort

J Clin Oncol

Human papillomavirus and survival of patients with oropharyngeal cancer

N Engl J Med

From monocytes to M1/M2 macrophages: phenotypical vs functional differentiation

Front Immunol

Infiltration of tumor-associated macrophages in human oral squamous cell carcinoma

Oncol Rep

CD163: a specific marker of macrophages in paraffin-embedded tissue samples

Am J Clin Pathol

Cancer-associated fibroblasts and CD163-positive macrophages in oral squamous cell carcinoma: their clinicopathological and prognostic significance

J Oral Pathol Med

Molecular mechanisms of HLA class I-mediated immune evasion of human tumors and their role in resistance to immunotherapies: HLA class I-mediated immune escape in tumors

HLA

Original article
Influence of tumor-associated macrophages and HLA class I expression according to HPV status in head and neck cancer patients receiving chemo/bioradiotherapy