头颈鳞癌的精准医疗研究进展

头颈鳞癌（head and neck squamous cell carcinoma,HNSCC）在全球范围内的发病率呈逐年上升趋势,尽管极力提高治疗水平,但其预后仍不理想。精准医疗时代,新的基因测序技术、液体活检技术、高分辨率成像技术以及分子病理学的进展,为HNSCC的精准诊断提供了良好的平台。而基于高通量基因测序及大数据分析等手段开发的精准细胞免疫治疗、个性化抗体偶联药物以及靶向人体微生物组药物等,为HNSCC的精准治疗提供了可靠的手段。作者综述了HNSCC精准诊断和精准治疗的相关研究进展。     

液体活检在头颈鳞癌中的研究和应用

头颈鳞癌是世界上最常见的恶性肿瘤之一,其早期诊断是迅速治疗并提高患者生存率的关键。液体活检主要包括循环肿瘤细胞、循环肿瘤DNA和外泌体的分析检测。与传统的影像学方法相比,液体活检主要通过非侵入性手段使疾病在早期时就被发现,并可通过连续取样来监测肿瘤发展进程。作者主要介绍液体活检的检测技术及其在头颈鳞癌中应用的最新研究进展。     

Photodynamic therapy in the management of pre-malignant head and neck mucosal dysplasia and microinvasive carcinoma

The management of head and neck mucosal dysplasia and microinvasive carcinoma is an appealing strategy to prevent the development of invasive carcinomas. While surgery remains the standard of care, photodynamic therapy (PDT) offers several advantages including the ability to provide superficial yet wide field mucosal ablative treatment. This is particularly attractive where defining the extent of the dysplasia can be difficult. PDT can also retreat the mucosa without any cumulative fibrotic complications affecting function. To date, clinical experience suggests that this treatment approach can be effective in obtaining a complete response for the treated lesion but long term follow-up is limited. Further research efforts are needed to define not only the risk of malignant transformation with PDT but also to develop site specific treatment recommendations that include the fluence, fluence rate and light delivery technique.     

The use of photodynamic therapy using 630 nm laser light and porfimer sodium for the treatment of oral squamous cell carcinoma

PDT has been demonstrated in clinical studies to be an efficacious method for the treatment of dysplastic, microinvasive and early forms of cancer. The advantage of PDT for early carcinomas of the oral cavity is the ability to preserve normal tissues while effectively treating cancers up to 1cm in depth. The case presented here successfully demonstrates the ability to use PDT to treat maxillary gingival squamous cell carcinoma thereby sparing the use of surgery or radiation therapy at this point in the management of the disease.     

趋化因子CXCL12及其受体CXCR4在头颈部鳞癌中的表达和临床意义

目的研究趋化因子CXCL12及其受体CXCR4在头颈部鳞癌中的表达情况及其与分化程度、侵袭性及淋巴结转移的关系。方法采用免疫组化方法检测96例头颈部鳞癌和20例头颈部正常上皮组织中CXCL12和CXCR4蛋白的表达情况及其与临床病理特征的关系。结果免疫组化结果发现CXCL12蛋白在头颈部鳞癌组织细胞中阳性表达率为68.8%,CXCR4蛋白在头颈部鳞癌组织细胞中阳性表达率为66.7%。CXCL12及其CXCR4在头颈部正常上皮组织中的表达为阴性。CXCR4的表达与T分期及N分期呈正相关(P<0.05),CXCR12的表达与T分期及N分期无关;CXCL12和CXCR4在头颈部鳞癌组织中的表达与肿瘤分化程度、肿瘤位置、患者年龄及性别无关。结论头颈部鳞癌组织中CXCL12和CXCR4蛋白的表达程度明显高于头颈部正常上皮组织,提示CXCR4-CXCL12轴在头颈部鳞癌发生、发展中发挥作用;CRCR4的表达与头颈部鳞癌局部浸润及淋巴结转移呈正相关,提示CXCR4在头颈部鳞癌组织中的表达可成为头颈部鳞癌预后不良的因素之一。     

外放疗联合个体化导板辅助 125I放射性粒子植入治疗晚期头颈部鳞癌

目的：探讨外放疗联合个体化导板辅助¹²⁵I放射性粒子植入治疗无法手术的头颈部鳞癌的可行性及不良反应。 方法：对17例因全身系统疾病而无法手术的头颈部鳞癌患者进行常规分割放疗联合个体化导板辅助¹²⁵I放射性粒子植入增量放疗。外放疗总照射剂量为50 Gy,放射性粒子匹配周缘剂量（matched peripheral dose, MPD）为80~120 Gy。 结果：通过肿瘤体积变化情况和临床表现评价治疗标准,17例患者治疗总有效率达到82.4%（14/17）,1例死于大出血。主要的不良反应包括：软组织坏死、局部出血及放射性黏膜炎和皮炎等,治疗过程未发生急性并发症与严重的放射性损伤。 结论：外放疗联合个体化导板辅助¹²⁵I放射性粒子植入治疗晚期头颈部鳞癌,是一种微创、有效、安全性高的治疗方案。     

Diffusion-weighted magnetic resonance imaging of head and neck squamous cell carcinomas

Objective

To evaluate whether diffusion-weighted imaging (DWI) is a reliable technique to quantify microstructural differences between head and neck squamous cell carcinomas (SCC) and tumour-free soft tissue.

Materials and methods

DWI was obtained from 20 patients with histologically proven, untreated head and neck SCC. DWI was acquired using a diffusion-weighted, navigated echo-planar imaging sequence with a maximum b-value of 800 s/mm². For an objective assessment of image quality, the signal-to-noise ratio (SNR) was calculated. Microstructural differences between vital tumour tissue and tumour-free soft tissue were quantified by calculating the apparent-diffusion-coefficients (ADC) on a pixel by pixel method.

Results

Echo-planar DWI provided good image quality in all patients (mean SNR 18.4). The mean ADC of SCC, (0.64 ± 0.28 × 10⁻³ mm²/s), was significantly (P < 0.0001) lower than that of the tumour-free soft tissue, (2.51 ± 0.82 × 10⁻³ mm²/s).

Conclusion

DWI is a reliable diagnostic tool to quantify the microstructural differences between vital tumour tissue and tumour-free soft tissue in patients with head and neck SCC.     

Potency of different red light sources in photodynamic induction of cell death in a squamous cell carcinoma cell line

LED illumination systems were found to be more efficacious than broad spectrum lamps in recent phase III trials on photodynamic treatment of actinic keratosis. However, a detailed comparison of the light doses emitted at the appropriate spectral range and its correlation to photodynamic effects is thus far not available for the most frequently used devices. Here, we compared the spectral emissions of three different PDT lamps with their potency of inducing cell death in ALA-loaded A431 cells, including a new system equipped with more advanced LEDs matching the photosensitizer absorption peak more precisely and emitting more homogeneous light over time. Cells were exposed to two different ALA concentrations, incubated for 1 or 3 h and then illuminated by one of two different LED or a broad-spectrum system at four different light doses, whereupon viability was assessed. Maximal doses were selected in accordance to clinically applied light doses in recent phase III studies and the manufacturers’ recommendations. The data gathered here clearly demonstrate that the two LED systems were significantly more effective in inducing cell death than the broad spectrum system. Most efficient was the newer LED system, in agreement with emission parameters that more accurately corresponded to the photosensitizer’s absorption peak.     

Residual tumour detection in post-treatment granulation tissue by using advanced diffusion models in head and neck squamous cell carcinoma patients

PurposeTo evaluate the detectability of the residual tumour in post-treatment granulation tissue using parameters obtained with an advanced diffusion model in patients with head and neck squamous cell carcinoma (HNSCC) treated by chemoradiation therapy.Materials and methodsWe retrospectively evaluated 23 patients with HNSCC after the full course of chemoradiation therapy. The diffusion-weighted image (DWI) acquisition used single-shot spin-echo echo-planar imaging with 11 b-values (0–1000). We calculated 10 DWI parameters using a mono-exponential model, a bi-exponential model, a stretched exponential model (SEM), a diffusion kurtosis imaging (DKI) model and a statistical diffusion model (SDM) in the region of interest (ROI) placed on the post-treatment granulation tissue. The presence of residual tumour was determined by histological findings or clinical follow-up.ResultsAmong the 23 patients, seven patients were revealed to have residual tumour. The univariate analysis revealed significant differences in six parameters between the patients with and without residual tumour. From the receiver operating characteristic curve analysis, the highest area under curve was detected in the center of the Gaussian distribution of diffusion coefficient (D_s) obtained by the SDM. The multivariate analysis revealed that the D_s and diffusion heterogeneity (α) obtained by the SEM were predictors for the presence of residual tumour.ConclusionDWI parameters obtained by advanced fitting models will be one of the diagnostic tools for the detection of residual tumour.     

Head and neck squamous cell carcinoma: usefulness of diffusion-weighted MR imaging in the prediction of a neoadjuvant therapeutic effect

The purpose of our study was to evaluate the usefulness of diffusion-weighted imaging in predicting the responses to neoadjuvant therapy for head and neck squamous cell carcinomas. Diffusion-weighted, T2-weighted, and gadolinium-enhanced T1-weighted images were obtained from 28 patients with untreated head and neck squamous cell carcinomas with histological proof. A blinded radiologist evaluated the quantitative and qualitative signal intensities and apparent diffusion coefficients (ADCs) in the lesions on each sequence. All patients were treated by neoadjuvant therapies, and the post-therapeutic tumor regression rate was determined. Both the quantitative and qualitative signal intensities on diffusion-weighted images showed positive correlations (r = 0.367 and 0.412, p < .05), and the ADCs showed a weak, inversed correlation (r = −0.384, p < .05) with the tumor regression rates. Diffusion-weighted imaging including an assessment by ADCs may be able to predict tumor response to neoadjuvant therapy for head and neck squamous cell carcinomas.     

Influence of photodynamic therapy on peripheral immune cell populations and cytokine concentrations in head and neck cancer

BackgroundPhotodynamic therapy (PDT) represents a palliative treatment option for a selected group of patients with head and neck squamous cell carcinoma (HNSCC). PDT induces a local inflammatory reaction with the potential to initiate antitumor immune responses. However, the systemic impact on peripheral immune cells has not been described so far.MethodsHNSCC patients (n = 9) were treated with PDT in a palliative setting. All patients had previously undergone several oncologic treatment regimens. Blood samples were taken before, during and after PDT. Age-matched healthy donors served as control group (NC, n = 15). The frequency and absolute number of T- and B-lymphocytes, CD4⁺CD39⁺ regulatory T-cells (Treg) and NK-cells were measured by 10-color flow cytometry. Serum concentrations of T cell related cytokine panel, including HMGB1, IL-6, IL-10 and perforin were measured by bead array and ELISA.ResultsIn heavily pretreated HNSCC patients, the number and frequency of Treg and NK-cells were increased as compared to NC. PDT induced a further increase of the frequency of Treg and NK-cells in the peripheral blood. Additionally, the serum concentrations of HMGB1, IL-6 and IL-10 showed a significant elevation after treatment with simultaneously decreased perforin levels.ConclusionAlthough PDT is a local treatment regimen, a systemic inflammatory response with altered peripheral immune cell populations and cytokine concentrations is visible. The increased Treg and NK cell numbers after PDT support the hypothesis that PDT may successfully be combined with NK cell or T cell activating immune checkpoint modulators in HNSCC patients to improve HNSCC specific immunity.     

Effects of photodynamic therapy with talaporfin sodium on squamous cell carcinoma and sarcoma cells

BackgroundPhotodynamic therapy (PDT) uses a photosensitizer and light to destroy abnormal cells. Talaporfin sodium (NPe6) is a second-generation photosensitizer.MethodsWe evaluated the toxic effects of different combinations of laser and NPe6 doses on squamous cell carcinoma (KLN205) and sarcoma (Meth A) cell lines. The cells were incubated with 0, 5, 10, or 30 μg/mL NPe6 for 24 h. The cells were then irradiated with 0, 5, 15, or 30 mW/cm² of laser power, and 0, 1, 5, 10, or 20 J/cm² of laser energy. Cell viability was evaluated after 24 h. We also evaluated the cytotoxic effects of continuous wave or square-wave modulated laser irradiations (2, 5, or 10 Hz, 50% duty) on Meth A cells.ResultsThe median lethal doses of NPe6 against the KLN205 and Meth A cells after treatment at a fluence rate of 15 mW/cm² and a light dose of 20 J/cm² were 18.6 and 5.0 μg/mL, respectively. Meth A cells were more sensitive to PDT than KLN205 cells. There was no significant difference between the effects of continuous wave and square-wave modulated lasers on Meth A cell viability.ConclusionsNPe6 PDT induced cell death in a dose-dependent manner in KLN205 and Meth A cells. More work is required to evaluate the cytotoxic effects of square-wave modulated laser therapy at low light doses.     

Hyaluronic acid carrier-based photodynamic therapy for head and neck squamous cell carcinoma

PurposeConventional photosensitizers for photodynamic therapy (PDT) typically have wide tissue distribution and poor water solubility. A hyaluronic acid (HA) polymeric nanoparticle with specific lymphatic uptake and highly water solubility was developed to deliver pyropheophorbide-a (PPa) for locally advanced head and neck squamous cell carcinoma (HNSCC) treatment.Methods and ResultsPPa was chemically conjugated to the HA polymeric nanoparticle via an adipic acid dihydrazide (ADH) linker. The conjugates were injected subcutaneously in a region near the tumor. Near-infrared (NIR) imaging was used to monitor distribution, and diode laser was used to activate PPa. The singlet oxygen generation efficiency of PPa was not affected by conjugation to HA nanoparticles at a PPa loading degree of 1.89 w.t.%. HA-ADH-PPa inhibited human HNSCC MDA-1986 cell growth only when photo-irradiation was applied. After HA-ADH-PPa treatment and radiation, NU/NU mice bearing human HNSCC MDA-1986 tumors showed reduced tumor growth and significantly enhanced survival time compared with an untreated group (p < 0.05).ConclusionsThese results demonstrate that HA-ADH-PPa could be useful for in vivo locoregional photodynamic therapy of HNSCC.     

<Superscript>123</Superscript>I-Interleukin-2 uptake in squamous cell carcinoma of the head and neck carcinoma

Introduction Information obtained on the IL-2 receptor status of tumour infiltrating lymphocytes in patients suffering from squamous cell carcinoma of the head and neck (SSCHN) before and after IL-2 treatment may lead to a better understanding of the immunological changes and related kinetics induced at the tumour level and ultimately to a strategy that allows selection of those patients that will benefit from IL-2 therapy. This study set out to assess the relationship between ¹²³I-IL2 single-photon emission computed tomography (SPECT) findings and the presence of IL-2 receptors (CD25 staining) on tumour-infiltrating lymphocytes as well as on SCCHN tumour cells in patients suffering from SCCHN. Materials and methods Seventeen consecutive patients (12 men; mean age, 57 years) highly suspected to suffer from SSCHN were prospectively included in the study. All patients underwent planar and whole body ¹²³I-IL2 scintigraphy and underwent surgery or had a biopsy taken within 1 week from imaging. Surgical resected primary lesions as well as biopsy material from primary tumours were histologically analysed with respect to the presence and intensity of CD25 expression on tumour infiltrating lymphocytes and tumour cells (HSCORE). Tumor-to-background (T/N) ratios of the primary tumour derived from planar and tomographic ¹²³I-IL2 scintigraphy were related to the results derived from histology. Results All patients suffered from SSCHN. T/N ratios derived from SPECT images were significantly correlated with CD25 lymphocyte HSCOREs (r = 0.66; p = 0.03), but not with CD25 tumour cell HSCOREs. Conclusions ¹²³I-IL-2 SPECT imaging allows for non-invasive imaging of the relative amount of IL-2 receptors present on tumour infiltrating lymphocytes in SCCHN.     

Clinical values for abnormal F-FDG uptake in the head and neck region of patients with head and neck squamous cell carcinoma

Purpose

Fluorine 18-fluorodeoxyglucose (¹⁸F-FDG) positron emission tomography (PET)/computed tomography (CT) is used to identify index or second primary cancer (SP) of the head and neck (HN) through changes in ¹⁸F-FDG uptake. However, both physiologic and abnormal lesions increase ¹⁸F-FDG uptake. Therefore, we evaluated ¹⁸F-FDG uptake in the HN region to determine clinical values of abnormal tracer uptake.

Methods

A prospective study approved by the institutional review board was conducted in 314 patients with newly diagnosed HN squamous cell carcinoma (HNSCC) and informed consent was obtained from all enrolled patients. The patients received initial staging workups including ¹⁸F-FDG PET/CT and biopsies. All lesions with abnormal HN ¹⁸F-FDG uptake were recorded and most of those were confirmed by biopsies. Diagnostic values for abnormal ¹⁸F-FDG uptake were calculated.

Results

Abnormal ¹⁸F-FDG uptake was identified in primary tumors from 285 (91.9%) patients. False-negative results were obtained for 22.3% (23/103) T1 tumors and 2.2% (2/93) T2 tumors (P < 0.001). Thirty-eight regions of abnormal ¹⁸F-FDG uptake were identified in 36 (11.5%) patients: the thyroid (n = 13), maxillary sinus (n = 7), palatine tonsil (n = 6), nasopharynx (n = 5), parotid gland (n = 2) and others (n = 5). Synchronous SP of the HN was identified in eight (2.5%) patients: the thyroid (n = 5), palatine tonsil (n = 2), and epiglottis (n = 1). The sensitivity and specificity of ¹⁸F-FDG PET/CT for identification of SPs were 75.0% and 98.7%, respectively.

Conclusions

¹⁸F-FDG PET/CT is a reliable method for tumor staging and identify SP in HN region, promoting appropriate therapeutic planning. Additional examinations may be required to identify superficial or small-volume tumors.     

Transcytolemmal water exchange in pharmacokinetic analysis of dynamic contrast-enhanced MRI data in squamous cell carcinoma of the head and neck

PURPOSE: To investigate the effect of transcytolemmal water exchange on the dynamic contrast-enhanced (DCE) T(1)-weighted MRI of human squamous cell carcinomas of the head and neck (HNSCC). MATERIALS AND METHODS: Nine patients with HNSCC nodal metastasis underwent pretreatment DCE-MRI with a temporal resolution of 2.5 seconds and a spatial resolution of 1 mm x 1 mm x 5 mm at 1.5T. We used two pharmacokinetic models for data analysis: generalized kinetic model (GKM) without considering transcytolemmal water exchange and the shutter-speed model (SSM), based on a two-site exchange model for transcytolemmal water exchange. The results were compared in three subgroups of voxels in the tumor depending on the level of contrast enhancement. RESULTS: SSM was found to be a better fit for more than 75% of pixels of all subjects (P < 0.01) in terms of residual size and Bayesian information criterion (BIC). For all three subgroups based on the contrast enhancement, the median K trans values of SSM were 42% to 55% higher than those of GKM and the median upsilon e values of SSM were 116% to 176% larger than those of GKM. The median K trans and upsilon e of two models were found significantly different (P < 0.01). The median tau i measured by SSM were from 211 to 364 msec. CONCLUSION: The effect of transcytolemmal water exchange is an important factor that needs to be incorporated for adequate modeling of contrast enhancement dynamics measured by MRI of HNSCC.     

Photodynamic therapy activated STAT3 associated pathways: Targeting intrinsic apoptotic pathways to increase PDT efficacy in human squamous carcinoma cells

5-Aminolaevulinic acid-based photodynamic therapy (ALA-PDT) has been used for part of squamous cell carcinoma (premalignant conditions or in situ cutaneous SCC–Bowen disease). However, mechanism of ALA-PDT is not fully understood yet on the cell apoptosis pathway. The aim of this study was to further investigate the effect and mechanism of 5-ALA-PDT on human squamous carcinoma A431cells. Apoptosis and cell viability after PDT were evaluated using Annexin V-FITC apoptosis detection kit and MTT assay. The mRNA and protein levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Our data showed that 5-ALA-PDT significantly inhibited cell proliferation (p< 0.05), but there was no significant difference when the photosensitizer reached to 4.8 mM. The inhibition in cell proliferation after 5-ALA-PDT treatment was correlated to more cells being arrested in the G0/G1 phase of the cell cycle (p < 0.01). Immunocytochemical observations using anti-active caspase-3 antibodies showed active caspase-3 was translocated from cytoplasm to nuclear during apoptosis. STAT3 and its downstream gene Bax and BCL-2 were changed after 5-ALA-PDT treatment for the mRNA and protein expression. Our studies confirmed that 5-ALA-PDT might be an effective treatment for human squamous carcinoma by inhibiting the tumor cell A431growth and for the first time demonstrated that the expression of STAT3 was significantly reduced at 24 h after 5-ALA-PDT treatment.