Advanced Laryngeal Squamous Cell Carcinoma Research Articles

Retrospective assessment of neoadjuvant camrelizumab combined with induction chemotherapy: efficacy in laryngeal preservation for advanced hypopharyngeal and laryngeal squamous cell carcinoma

BackgroundHypopharyngeal and laryngeal squamous cell carcinoma (SCC) account for 25–30% of head and neck SCC. Total laryngectomy, while effective, compromises the quality of life. Immune checkpoint inhibitors such as Camrelizumab offer potential in laryngeal preservation. The study investigated Camrelizumab combined with TP regimen as a neoadjuvant therapy for laryngeal preservation in advanced hypopharyngeal and laryngeal SCC.MethodsA retrospective study was conducted at Sun Yat-sen University Cancer Center on patients diagnosed with locally advanced SCC of the hypopharynx and larynx from October 1, 2019, to October 25, 2022. The efficacy of a first-line treatment combining Camrelizumab (200 mg) and TP regimen (Albumin-bound paclitaxel at 260 mg/m2 and Cisplatin at 60 mg/m2) was evaluated using RECIST 1.1 criteria. Outcomes included overall survival (OS), progression-free survival (PFS), laryngectomy-free survival (LFS), and response rates.ResultsOf the 71 included patients, the median age was 60.7 years. Post the first-line treatment, 90.1% demonstrated an overall response. The one-year and two-year OS rates were 91.5% and 84.3%, respectively. One-year and two-year PFS rates were 92.9% and 83.9%, respectively, with LFS at 85.6% and 73.2%. The initial T4 stage as significantly associated with reduced OS and LFS. Skin reaction was the predominant adverse event.ConclusionThe Camrelizumab-TP regimen demonstrated promising results for advanced hypopharyngeal/laryngeal SCC patients, exhibiting high response rates, OS, and LFS, positioning it as a potential primary option for laryngeal preservation. Further comprehensive, randomized controlled studies are imperative to validate these initial observations and elucidate the regimen's full clinical efficacy in optimizing laryngeal outcomes.

Chemoradiotherapy vs radiotherapy for non-surgical locally advanced laryngeal squamous cell carcinoma patients: a propensity score-matched study and practical nomogram construction.

To compare the cancer-specific survival (CSS) among patients with locally advanced laryngeal squamous cell carcinoma (LSCC) receiving chemoradiotherapy (CRT) and radiotherapy (RT) treatment, as well as to establish a prognostic nomogram for survival prediction in patients receiving CRT. Using data from the Surveillance, Epidemiology, and End Results (SEER) database, patients with laryngeal cancer were identified between 2010 and 2015, with follow-up up to 2018. Propensity score matching (PSM) was performed to minimize disproportionate distributions of the potential confounding. Cox regression models were used to evaluate the CSS of two treatment groups. A prognostic nomogram for patients receiving CRT was then developed and evaluated. Totally 1085 non-surgical patients with locally advanced LSCC were included in this study (median [IQR] age, 62 [55-69] years; 829 [76.41%] males), of which 913 receiving CRT and 172 receiving RT. After PSM, significantly improved CSS was observed in locally advanced LSCC patients receiving CRT when compared to RT (HR: 0.62 [95% CI 0.42-0.92]; P = 0.014). Then, in the group of 639 locally advanced LSCC patients receiving CRT, a prognostic nomogram based on age, tumor size, N category, and marital status were developed and validated, of which the predictive performance was superior to that of TNM staging system (7th edition). CSS shows a statistically significant improvement in locally advanced LSCC patients who receipt of CRT when compared with RT. Furthermore, a prognostic nomogram for locally advanced LSCC patients receiving CRT was established, which shows a good calibration and identification accuracy.

Abstract C068: Exploring the relationship between miR-9-5p levels and MAP1B expression in laryngeal cancer cell lines

Abstract Background: Low miR-9-5p expression has been associated with poor overall survival in head and neck squamous cell carcinoma (HNSCC), whereas higher expression is associated with better outcomes. Our previous studies determined that low miR-9 levels is a significant feature of advanced stage laryngeal squamous cell carcinoma (LSCC) specifically in Black patients. We further showed that low miR-9 is associated with increased cell proliferation and decreased chemosensitivity to cisplatin. In this study, we sought to explore the relationship between low miR-9-5p- and high miR-9-5p-LSCC cell lines and its genomic and potential chemotherapeutic target, microtubule-associated protein 1B (MAP1B). Methods: We used four cell lines derived from patients with advanced stage LSCC: UM-SCC-98, UM-SCC-12, UM-SCC-11A, UM-SCC-10A (University of Michigan). Mature miR-9-5p levels were determined in each cell line via northern blotting. Baseline chemosensitivity (IC50 of cisplatin) as well as cell proliferation of each cell line was assessed using the Cell Titer Blue Viability assay. Baseline MAP1B gene and protein expression were determined in each cell line via qPCR and western blotting, respectively. Results: The lowest mature miR-9-5p levels were found in UM-SCC-98 followed by UM-SCC-12. The highest mature miR-9-5p levels were found in UM-SCC-11A followed by UM-SCC-10A. Cell lines with lower miR-9-5p resulted in increased cell proliferation and higher cisplatin IC50 (p < 0.0001): UM-SCC-98 IC50 = 18.32µM and UM-SCC-12 IC50 = 16.70µM. Conversely, cell lines with higher miR-9-5p levels resulted in lower cell proliferation and lower cisplatin IC50; UM-SCC-11A IC50 = 2.28µM and UM-SCC-10A IC50 = 10.95µM. An inverse relationship between miR-9-5p levels and MAP1B gene expression was seen, wherein the low miR-9-5p expressing cell lines (UM-SCC-98 and UM-SCC-12) had the highest gene expression levels; the high miR-9-5p expressing cell lines (UM-SCC-11A and UM-SCC-10A) had the lowest gene expression levels (p < 0.0001, fold change >2). Lower miR-9-5p levels also corresponded with higher MAP1B protein expression, wherein expression was only seen in cell lines UM-SCC-98 and UM-SCC-12. No MAP1B protein expression was seen in UM-SCC-11A and UM-SCC-10A (p < 0.0001). Conclusion: This study further supports the relevance of low miR-9-5p in the regulation of cisplatin chemosensitivity in LSCC. MAP1B, a described mediator of chemoresistance, may be a future target for intervention. Future studies will involve exploring mechanisms of miR-9-5p inhibition and defining MAP1B regulatory pathways. Citation Format: Christina Gobin, Chayil C. Lattimore, Jai Walker, Kristianna M. Fredenburg. Exploring the relationship between miR-9-5p levels and MAP1B expression in laryngeal cancer cell lines [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C068.

Thyroid cartilage infiltration in advanced laryngeal cancer: prognostic implications and predictive modelling.

Detection of laryngeal cartilage invasion is of great importance in staging of laryngeal squamous cell carcinoma (LSCC). The role of prognosticators in locally advanced laryngeal cancer are still widely debated. This study aimed to assess the impact of volume of thyroid cartilage infiltration, as well as other histopathologic variables, on patient survival. We retrospectively analysed 74 patients affected by pT4 LSCC and treated with total laryngectomy between 2005 and 2021 at the Department of Otorhinolaryngology - Head and Neck Surgery of the University of Brescia, Italy. We considered as potential prognosticators histological grade, perineural (PNI) and lympho-vascular invasion (LVI), thyroid cartilage infiltration, and pTN staging. Pre-operative CT or MRI were analysed to quantify the volume of cartilage infiltration using 3D Slicer software. The 1-, 3-, and 5-year disease free survivals (DFS) were 76%, 66%, and 64%, respectively. Using machine learning models, we found that the volume of thyroid cartilage infiltration had high correlation with DFS. Patients with a higher volume (> 670 mm3) of infiltration had a worse prognosis compared to those with a lower volume. Our study confirms the essential role of LVI as prognosticator in advanced LSCC and, more innovatively, highlights the volume of thyroid cartilage infiltration as another promising prognostic factor.

Predicting survival of advanced laryngeal squamous cell carcinoma: comparison of machine learning models and Cox regression models

Laryngeal squamous cell carcinoma (LSCC) is a common tumor type. High recurrence rates remain an important factor affecting the survival and quality of life of advanced LSCC patients. We aimed to build a new nomogram and a random survival forest model using machine learning to predict the risk of LSCC progress. The study included 671 patients with AJCC stages III–IV LSCC. To develop a prognostic model, Cox regression analyses were used to assess the relationship between clinic-pathologic factors and disease-free survival (DFS). RSF analysis was also used to predict the DFS of LSCC patients. The ROC curve revealed that the Cox model exhibited good sensitivity and specificity in predicting DFS in the training and validation cohorts (1 year, validation AUC = 0.679, training AUC = 0.693; 3 years, validation AUC = 0.716, training AUC = 0.655; 5 years, validation AUC = 0.717, training AUC = 0.659). Random survival forest analysis showed that N stage, clinical stage, and postoperative chemoradiotherapy were prognostically significant variables associated with survival. The random forest model exhibited better prediction ability than the Cox regression model in the training cohort; however, the two models showed similar prediction ability in the validation cohort.

PD-1 Inhibitors combined with paclitaxel (Albumin-bound) and cisplatin for larynx preservation in locally advanced laryngeal and hypopharyngeal squamous cell carcinoma: a retrospective study.

laryngeal and hypopharyngeal squamous cell carcinoma (SCC) is a common head and neck cancer with significant impact on quality of life due to its crucial roles in vocalization, airway protection, and swallowing. This retrospective study aims to evaluate the efficacy and larynx organ preservation of neoadjuvant treatment with PD-1 inhibitors in combination with paclitaxel (Albumin-bound) and cisplatin for locally advanced laryngeal and hypopharyngeal SCC. Medical records of consecutive patients diagnosed with histologically or cytologically confirmed locally advanced SCC of the larynx and hypopharynx, who received PD-1 inhibitor therapy at a single tertiary care center, were reviewed from January 1, 2019, to December 15, 2022. The patients were treated with a combination of PD-1 inhibitors, paclitaxel (Albumin-bound) 260mg/m2, and cisplatin 60mg/m2 (TP) as their first-line therapy. Survival outcomes, laryngectomy-free survival (LFS) rates and response rates were assessed. The study cohort comprised 156 patients, predominantly male, with a median age of 60.4 years. The estimated one-year overall survival (OS) rate was 94.1%, two-year OS rate was 82.5%, one-year progression-free survival (PFS) rate was 80.4%, and two-year PFS rate was 66.3%. The one-year LFS was 86.4%, and the two-year LFS rate was 73.0%. The overall response rate after TP + PD-1 inhibitors therapy was 88.5%. Common treatment-associated adverse events included rash, thyroid function abnormalities, myelosuppression, and colitis. Neoadjuvant therapy with PD-1 inhibitors in combination with paclitaxel (Albumin-bound) and cisplatin showed promising efficacy and tolerability for larynx preservation in locally advanced laryngeal and hypopharyngeal SCC. The high response rates and favorable survival outcomes suggest this approach as a potential treatment option. Prospective randomized controlled trials are needed to further validate these findings and establish the role of immunotherapy in larynx preservation.

High-grade tumor budding is a risk factor for survival in patients with laryngeal squamous cell carcinoma

ObjectiveWith the increasing incidence and mortality of laryngeal squamous cell carcinoma worldwide, researchers continue to search for novel prognostic factors and treatment methods for preventing early laryngeal squamous cell carcinoma from becoming advanced laryngeal squamous cell carcinoma. This study aims to determine if tumor budding is an independent risk factor associated with the survival of patients with laryngeal squamous cell carcinoma. Methods268 cases of laryngeal squamous cell carcinoma were studied, and tumor budding was analyzed for associations with clinicopathological features and clinical outcomes. ResultsTumor budding was divided into low-grade tumor budding (0–6/0.785 mm2) and high-grade tumor budding (≥7/0.785 mm2) based on the results of the receiver operating characteristics curve analysis. Logistic regression analysis showed that smaller tumor cell nests, the low levels of tumor-infiltrating lymphocytes, and higher pathological T staging were the risk factors for high-grade tumor budding (p < 0.05). In the low-grade tumor budding group, there was no statistic difference in survival between patients without tumor budding and those with 1–6/0.785 mm2 tumor budding. Multivariate survival analysis showed high-grade tumor budding (p < 0.001) was independent prognostic factors for disease-free survival and overall survival in laryngeal squamous cell carcinoma. High-grade tumor budding was also an independent prognostic factor for disease-free survival (p = 0.037) and overall survival (p = 0.009) in T1-2N0 laryngeal squamous cell carcinoma. ConclusionsSmaller tumor cell nests, the low levels of tumor-infiltrating lymphocytes, and higher pathological T staging were closely associated with high-grade tumor budding in laryngeal squamous cell carcinoma. High-grade tumor budding may be an adverse risk factor that affects not only the disease-free survival and overall survival of laryngeal squamous cell carcinoma patients but also the survival of T1-2N0 laryngeal squamous cell carcinoma patients. Level of EvidenceLevel 4.

Locally Advanced Laryngeal Squamous cell Carcinoma in a Patient with a Synchronous Second Primary Thyroid Carcinoma

Locally Advanced Laryngeal Squamous cell Carcinoma in a Patient with a Synchronous Second Primary Thyroid Carcinoma

Neoadjuvant chemoimmunotherapy for the treatment of locally advanced laryngeal and hypopharyngeal squamous cell carcinoma: A single-arm phase 2 clinical trial.

6077 Background: Standard treatment for patients with locally advanced laryngeal and hypopharyngeal squamous cell carcinoma (LHSCC) is total laryngectomy, which seriously affects the quality of life. Neoadjuvant treatment with Nab-paclitaxel plus cisplatin(TP) has favorable efficacy with acceptable toxicity for laryngeal preservation in LHSCC. Programmed death 1 (PD-1) blockade plus chemotherapy has shown a survival benefit and recommended as the first-line treatment in recurrent or metastatic head and neck cancer, but the safety and activity in the combination of TP and PD-1 blockade in locally advanced LHSCC need investigation. Methods: In this single-center, single-arm, phase 2 trial, newly diagnosed pts with stage T3-4N0-3M0 laryngeal SCC or T2-4N0-3M0 hypopharyngeal SCC were recruited. Eligible patients received chemotherapy [Nab-paclitaxel 240 mg/m2 plus cisplatin 75 mg/m2] and toripalimab[240 mg, d1]every 3 weeks for 3 cycles. After induction treatment, patients achieving clinical partial response (cPR) or complete response (cCR) received definitive radiotherapy, while those with stable diseases(SD) and progression disease(PD) received surgery plus adjuvant radiotherapy. Toripalimab was given intravenously once every 3 weeks for up to 10 cycles, including 3 cycles before. The primary endpoint was the objective response rate (ORR) according to RECIST 1.1 by investigator assessment to neoadjuvant chemoimmunotherapy. The secondary endpoints included laryngectomy-free survival (LFS), disease failure-free survival (DFS)and overall survival OS at 2 years, quality of life (QOL) and toxic effects. Results: From October 2020 to May 2022，25 patients received neoadjuvant chemoimmunotherapy (median age 58 years; 23.9% men), 52% (13/25) and 48% (12/25) were hypopharyngeal and laryngeal squamous cell carcinoma. 80% (20/25) were clinical T3/4 and 64.0% (16/25) ≥N2. After neoadjuvant chemoimmunotherapy, 2 patients achieved CR, 21 achieved PR, with an ORR of 92%(23/25), 23 patients with CR/PR of the primary tumor received concurrent radiotherapy and immune maintenance therapy, 2 patients with SD/PD received laryngectomy. At a median follow-up of 17 months (range 8-27 months), 1-year DFS was 88.0% , 1-year OS was 96.0% , 1-year LPS was 92.0%, Respectively, only grade 1-2 adverse events occurred in 100% of patients, including loss of appetite (100%), anemia (92.0%), nausea (84.0%), vomiting (52.0%), hypothyroidism (16.0%), peripheral neuritis (20%), leukopenia (12.0%), rash (12.0%). Conclusions: Neoadjuvant treatment with TP plus toripalimab achieved impressive ORR and 1-year LPS rate with manageable toxicities in patients with LHSCC. Further follow-up is needed to confirm the long-term efficacy. Clinical trial information: ChiCTR2000033506 .

Expression and prognostic value of ING3 in advanced laryngeal squamous cell carcinoma

Objectives: Laryngeal squamous cell carcinomas (LSCC) is one of the most common aggressive neoplasms of the head and neck region. There is a significant need for identification of successful and accurate prognostic markers to better estimate the clinical outcomes for LSCC patients. In this study, we aimed at analyzing the differential expressions of inhibitor growth (ING) family members and to evaluate the prognostic values of deregulated ING genes in LSCC. Methods: We investigated the relative expressions of ING genes in laryngeal tumor-normal tissue pairs in the mRNA level using quantitative real-time polymerase chain reaction and relative expression of ING3 in the protein level using Western Blot analysis. Results: The rate of genetic alterations of ING3 was relatively higher in head and neck cancer including LSCC. ING3 expression was significantly upregulated in LSCC tissue samples in both mRNA and protein level. Higher expression of ING3 was also correlated with poor disease-free survival of patients with head and neck cancer. Conclusions: Our findings assigned an oncogenic feature for ING3 in laryngeal cancer with a significant upregulation detected in advanced cases and suggested a vital prognostic potential for ING3.

The relation between hypoxia and proliferation biomarkers with radiosensitivity in locally advanced laryngeal cancer

PurposeTreatment decision-making in advanced-stage laryngeal squamous cell carcinoma (LSCC) is difficult due to the high recurrence rates and the desire to preserve laryngeal functions. New predictive markers for radiosensitivity are needed to facilitate treatment choices. In early stage glottic LSCC treated with primary radiotherapy, expression of hypoxia (HIF-1α and CA-IX) and proliferation (Ki-67) tumour markers showed prognostic value for local control. The objective of this study is to examine the prognostic value of tumour markers for hypoxia and proliferation on locoregional recurrent disease and disease-specific mortality in a well-defined cohort of patients with locally advanced LSCC treated with primary, curatively intended radiotherapy.MethodsIn pre-treatment biopsy tissues from a homogeneous cohort of 61 patients with advanced stage (T3–T4, M0) LSCC primarily treated with radiotherapy, expression of HIF-1α, CA-IX and Ki-67 was evaluated with immunohistochemistry. Demographic data (age and sex) and clinical data (T- and N-status) were retrospectively collected from the medical records. Cox regression analysis was performed to assess the relation between marker expression, demographic and clinical data, and locoregional recurrence and disease-specific mortality.ResultsPatients with high expression of HIF-1α developed significantly more often a locoregional recurrence (39%) compared to patients with a low expression (21%) (p = 0.002). The expression of CA-IX and Ki-67 showed no association with locoregional recurrent disease. HIF-1α, CA-IX and Ki-67 were not significantly related to disease-specific mortality. Clinical N-status was an independent predictor of recurrent disease (p < 0.001) and disease-specific mortality (p = 0.003). Age, sex and T-status were not related to locoregional recurrent disease or disease-specific mortality.ConclusionHIF-1α overexpression and the presence of regional lymph node metastases at diagnosis were independent predictors of locoregional recurrent disease after primary treatment with curatively intended radiotherapy in patients with locally advanced LSCC.

Higher Expression Levels of miR-1246 in Advanced Laryngeal Carcinoma Are Associated with Elevated Risk of Locoregional Metastasis

Worldwide, laryngeal squamous cell carcinoma (LSCC) is the second most common head and neck cancer (HNC). Advanced forms of LSCC, especially HPV-negative types, frequently involve locoregional lymph node metastasis, which are associated with treatment failure. Despite recent development and improvement in treatment protocols [1], still advanced LSCC is a global oncological burden, associated with challenges and complications. The study group included 60 cases of predominantly HPV-negative tumours with all patients having history of long-term smoking. We found that miR-1246 was significantly dysregulated in advanced LSCC – almost 72% of the patients in the study group showed overexpression of miR-1246 in tumour tissue above relative quantity of 2 (RQ &gt; 2). Additionally, miR-1246 showed significant heterogeneity in expression levels between tumour depth and surface and moreover higher expression levels of the molecule correlated with higher chance of locoregional metastasis. In conclusion, we explored the expression levels of miR-1246 in advanced LSCC and adjacent normal tissue, revealing new tumour heterogeneity characteristic. We demonstrated potential association between miR-1246 expression levels and presence of locoregional lymph nodal metastasis.

The value of preoperative inflammatory markers in patients through total laryngectomy for advanced stage larynx carcinoma

The aim of this study is to evaluate the difference between neutrophil, lymphocyte, platelet and RDW (red cell distribution width) counts and NLR (neutrophil/lymphocyte ratio) and PLR (platelet/lymphocyte ratio) between patients who underwent total laryngectomy for advanced stage laryngeal squamous cell carcinoma and healthy control group. The study group consisted of 37 patients (36 male, 1 female) who underwent total laryngectomy and bilateral neck dissection for advanced stage laryngeal squamous cell carcinoma between January 2017 and December 2019. As a control group, complete blood count records of 35 healthy individuals who were routinely examined in 2019 within the scope of occupational health and safety were compiled. Blood was collected preoperatively from the patient in the study group. The data obtained from the whole blood counts of the two groups were analyzed statistically by comparing neutrophil, lymphocyte, platelet, RDW, NLR and PLR values. The age difference between the study and control groups was not significant (p&gt;0.05). Neutrophil, platelet and RDW counts were higher in the study group compared to the control group and this difference was statistically significant (p=0.001, p=0.035, p=0.001, respectively). Although the lymphocyte count was high in the study group, the difference was not significant (p=0.061). The difference between NLR and PLR between the two groups was high in the study group and this difference was statistically significant (p=0.001, p=0.007, respectively). Neutrophil, platelet, RDW, NLR, PLR values were significantly higher in patients with advanced stage laryngeal cancer than in the control group. NLR, PLR and RDW values, which can be obtained cheap, simple and rapidly at the diagnosis stage of laryngeal cancers, may be biomarkers in determining prognosis. For this, multicenter, prospective studies are needed.

Treatment and outcome among patients with laryngeal squamous cell carcinoma in Stockholm-A population-based study.

Survival of patients with advanced laryngeal squamous cell carcinoma (LSCC) remains poor and management protocols warrant further development. We thus investigated treatment and outcome-related factors for LSCC in Stockholm, Sweden. In a retrospective setting, 520 patients with LSCC diagnosed during 2000-2014, were included. Data on stage, treatment, and outcome were correlated with recurrence-free and overall survival (RFS and OS, respectively). Five-year OS for all patients was 65%. Five-year RFS for T1a, T1b, T2, T3, and T4 glottic LSCC was 90%, 91%, 77%, 47%, and 80%, respectively. The corresponding figures for T1, T2, T3, and T4 supraglottic LSCC were 64%, 66%, 64%, and 86%. Patients with a T3 glottic LSCC had unexpectedly poor survival, especially when compared with patients with a T4 tumor. Patients with T4 disease were primarily treated with laryngectomy and postoperative radiotherapy (RT)/chemoradiotherapy (CRT), while most patients with T3 LSCC were treated with RT/CRT.

Abstract C042: Low miR-9 impacts tumorigenesis and chemoresistance in Black compared with White laryngeal squamous cell carcinoma

Abstract Background: Advanced stage laryngeal squamous cell carcinoma (LSCC) has sustained one of the lowest five-year survival rates across all other head and neck squamous cancers. For several decades, Black patients have maintained a greater likelihood of presenting with advanced LSCC and a higher overall mortality rate compared with White patients. Even when accounting for sociodemographics, a disparity remains, underscoring the importance of investigating the biologic basis of this disparity. Differential microRNA (miRNA) expression has been linked to racially disparate clinical outcomes in other cancers including breast and prostate, but remains underdetermined in LSCC. We present our findings where we investigated miR-9 levels and their influence on LSCC tumorigenesis and chemoresistance using in vitro cell lines derived from Black and White patients. Methods: We previously identified miR-9-5p as significantly lower (5X) in Black compared with White advanced stage LSCC in vivo samples. For functional studies, we obtained two LSCC cell lines derived from a Black patient (UM-SCC-12) and from a White patient (UM-SCC-10A) (provided by Tom Carey, University of Michigan). After transfection with either miR-9 mimic or inhibitor, we used the scratch wound assay to assess cell migration and the cell titer blue assay to assess cell proliferation and chemoresistance. Results: Prior to transfection, we found that UM-SCC-12 had baseline increases in cellular migration, proliferation and chemoresistance compared to UM-SCC-10A. By Northern blot analysis, we further determined that mature miR-9-5p was inherently 5X lower in UM-SCC-12 compared with UM-SCC-10A. After transient transfection, we noted that overexpressing miR-9 in UM-SCC-12 resulted in decreased cellular migration, decreased proliferation and increased chemosensitivity compared to a mock oligo control, whereas inhibiting miR-9 in UM-SCC-10A resulted in increased cellular migration, increased proliferation and decreased chemosensitivity compared to a mock oligo control. Conclusion: Collectively, these studies support that low miR-9 may play a role in LSCC tumorigenesis and chemoresistance. In this manner, it may contribute to cancer health disparate outcomes observed in advanced stage LSCC. Future studies will involve characterizing significant downstream mediators of low miR-9-5p gene regulation. Citation Format: Christina Gobin, Samuel Inkabi, Chayil Lattimore, James Menefee, Tengfei Bian, Christopher Fields, Mingyi Xie, Chengguo Xing, Kristianna Fredenburg. Low miR-9 impacts tumorigenesis and chemoresistance in Black compared with White laryngeal squamous cell carcinoma [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr C042.

A preliminary analysis of prognostic genes in advanced laryngeal squamous cell carcinoma patients with postoperative radiotherapy.

Advanced laryngeal squamous cell carcinoma (LSCC) has a high mortality rate, and the prognosis is poor. However, the underlying molecular biological mechanisms bringing about the development and progression of advanced LSCC are not entirely clarified. This study aimed to find out the potential biomarkers to predict the prognosis in advanced LSCC patients who had undergone postoperative radiotherapy alone. The next-generation sequencing of RNA was performed to detect the mRNAs expression profiling in 10 advanced LSCC samples, comprised of 5 samples from LSCC patients with favorable outcome and 5 samples from paired patients with poor outcome. Then bioinformatics analysis including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were used to find out functional core genes that were significantly different between the two groups. 1630 differentially expressed genes (DEGs) were confirmed to have significant differences between the two groups. 53 GO terms and 19 pathways which were closely related to the DEGs were identified. Finally, 52 intersection DEGs which were both related to the top three GO terms and pathways were identified. The expression of several core genes was confirmed with RT-qPCR in tissues from another 75 patients. RT-qPCR confirmed that the genes of c-JUN, LYN, PIK3R2, and TNFAIP3 were significantly differentially expressed between the two groups, which was in accordance with the RNA sequencing data. The DEGs identified above may be potential prognostic markers for advanced LSCC patients with postoperative radiotherapy, and may provide essential guidance for following-up.

Differentially expressed genes related to lymph node metastasis in advanced laryngeal squamous cell cancers.

Understanding the molecular mechanisms and gene expression in laryngeal squamous cell carcinoma (LSCC) may explain its aggressive biological behavior and regional metastasis pathways. In the present study, patients with locally advanced LSCC tumors were examined for differential gene expression in the normal mucosa (non-tumoral mucosa), tumors and lymph node tissues. The aim was to identify possible predictive genes for lymph node metastasis. A total of 16 patients who had undergone total laryngectomy with neck dissection for advanced LSCC were randomly selected from a hospital database: Eight of the patients had lymph node metastasis (Group 1) and the other eight patients did not have metastasis (Group 2). Overall survival (OS), disease-free survival (DFS) and disease-specific survival (DSS) were analyzed. For each patient, paraffin-embedded tissue samples were collected from non-tumoral mucosa, tumoral lesions and lymph node tissues. RNA was extracted from the tissue samples and used for complementary DNA synthesis, and microarray analysis was subsequently performed on each sample. Gene expression levels were determined in each specimen, and Groups 1 and 2 were compared and statistically analyzed. The microarray results for lymph node metastasis-positive and -negative groups, indicated the differential expression of 312 genes in the lymph nodes, 691 genes in the normal mucosal tissue and 93 genes in the tumor tissue. Transgelin (TAGLN) and cofilin 1 (CFL1) were identified as possible target genes and validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The RT-qPCR results for TAGLN and CFL1 supported the microarray data. OS, DFS and DSS times were longer in Group 2 than in Group 1 (P=0.002, 0.015 and 0.009, respectively). In addition, TAGLN and CFL1 were associated with DFS and DSS. On the basis of these results, it is suggested that TAGLN and CFL1 expression may play an important role in the pathogenesis of regional metastasis and poor prognosis in advanced LSCC.

The Exosomal miR-1246 of Laryngeal Squamous Cell Carcinoma Induces Polarization of M2 Type Macrophages and Promotes the Invasiveness of Laryngeal Squamous Cell Carcinoma.

Background The possible role and detailed mechanisms of Tumor-associated macrophages (TAMs) in laryngeal squamous cell carcinoma (LSCC) have not been revealed. Methods The expressions of typical markers were evaluated by qRT-PCR. In macrophages cocultured with TU212 cells, CD163, and CD206 protein expressions were detected by western blot analysis; IL-10 and IL-12 expressions were detected by ELISA assay. Exosomes isolated from TU212 cells were characterized by TEM analysis. As for the TU212 cells cocultured with macrophages processed with HOK or TU212 cells derived exosomes, their viability, migration, and invasion were assessed by CCK-8 assay, wounding healing, and Transwell assays, respectively. Results In this study, macrophages processed with exosomes from human TU212 cells notably advanced LSCC cell viability, migration, and invasion. miR-1246 inhibitor suppressed the M2 polarization of macrophages. Macrophages transfected with miR-1246 inhibitor suppressed LSCC cell viability, migration, and invasion. Conclusion In summary, our data implied that the exosomal, miR-1246 of LSCC, induced polarization of M2 type macrophages and promoted the progression of LSCC. This trial is registered with 2020-13.

Open Partial Laryngectomies: History of Laryngeal Cancer Surgery.

Historically, surgery was the first-choice therapy for early, intermediate and advanced laryngeal squamous cell carcinoma (LSCC). Partial laryngeal surgery has evolved in recent decades and was influenced by many historic events and the development of new technologies. Partial laryngectomies may be performed by open, endoscopic or transoral robotic approaches. In this historic paper, we describe the evolution of open partial laryngectomy techniques, indications and surgical outcomes. Since the first partial laryngectomy in 1788, many U.S., U.K. and European surgeons, including Henry Sands, Jacob da Silva Solis-Cohen and Theodor Billroth, performed this surgical procedure under local anesthesia for tuberculosis, cancer or syphilis. Partial laryngectomy gained reputation in the medical community in 1888 due to the laryngeal cancer and death of the prince of Prussia, Frederick III. Frederick III’s death represented the turning point in the history of partial laryngectomies, calling attention to the importance of semiotics, biopsy and early diagnosis in laryngeal cancers. Hemi-laryngectomy was indicated/proposed for lateral laryngeal tumors, while thyrotomy was indicated for cancers of the middle part of the vocal fold. The second landmark in the history of partial laryngectomies was the discovery of cocaine, novocaine and adrenaline and the related development of local anesthetic techniques, which, together with the epidemiological and hygienic advances of the 19th century, allowed for better perioperative outcomes. General anesthesia was introduced in the second part of the 20th century and further improved the surgical outcomes. The diagnosis of laryngeal cancer was improved with the development of X-rays and direct laryngoscopies. The 20th century was characterized by the development and improvement of vertical partial laryngectomy procedures and the development of horizontal partial laryngectomies for both supraglottic and glottic regions. The history and the evolution of these techniques are discussed in the present historical paper.

Bioradiotherapy with Cetuximab May Reduce the Risk of Neck Node Relapse in Locoregionally Advanced Laryngeal Glottic Carcinoma: May HER1-Profile Be Useful in the Bioselection of Patients?

The aim of the study was to evaluate survival in patients with advanced glottic laryngeal squamous cell carcinoma treated by bioradiotherapy (BioRT) with cetuximab and eventual salvage surgery (group A, n = 66) or upfront surgery (total laryngectomy or near-total laryngectomy) with or without postoperative radiotherapy (PORT) (group B, n = 66). The predictive role of HER1 expression in the bioselection of tumors was evaluated. Relapse-free (RFS), metastasis-free (MFS), overall (OS) survivals, salvageability, and rates of larynx preservation were analyzed. The two groups were balanced by propensity score method on their baseline characteristics. No significant differences in RFS and OS were found, while MFS results were significantly higher in group A (p = 0.04). Group A showed a 22% reduction in the probability of nodal metastasis (p = 0.0023), mostly in tumors with higher HER1 expression. The salvageability with TL at 3 years was 54% after prior BioRT and 18% after prior upfront NTL (p < 0.05). BioRT with cetuximab showed a reduction in the risk of lymph node relapse, particularly in the case of HER1 positive tumors, and it allowed to achieve a higher rate of functional larynx preservation and a higher salvageability compared with upfront surgery. HER1 analysis could be clinically useful in the bioselection of tumors that may benefit from BioRT with cetuximab, particularly in those with neck node metastatic propensity.