Squamous Cell Carcinoma Research Articles

GPD1L may inhibit the development of esophageal squamous cell carcinoma through the PI3K/AKT signaling pathway: bioinformatics analysis and experimental exploration

GPD1L may inhibit the development of esophageal squamous cell carcinoma through the PI3K/AKT signaling pathway: bioinformatics analysis and experimental exploration

LINC01614 Promotes Oral Squamous Cell Carcinoma by Regulating FOXC1

Background: Long non-coding RNAs (lncRNAs) are pivotal mediators during the development of carcinomas; however, it remains to be investigated whether lncRNAs are implicated in oral squamous cell carcinoma (OSCC). Methods: In this study, quantitative real-time PCR was conducted for detecting the expression of LINC01614 in OSCC cell lines. The biological functions of LINC01614 were assessed by loss- and gain-of-function experiments conducted both in vivo and in vitro. Cellular proliferation, migration, and invasion were investigated herein, and dual luciferase reporter assays were additionally performed to explore the relationships among LINC01614, miR-138-5p, and Forkhead box C1 (FOXC1). Results: The research presented herein revealed that OSCC cells express high levels of LINC01614. Functional experiments employing cellular and animal models demonstrated that LINC01614 knockdown repressed the malignant phenotypes of OSCC cells, including their growth, invasiveness, and migration. Further investigation revealed that LINC01614 absorbs miR-138-5p miRNA by functioning as a competing endogenous RNA to downregulate the abundance of FOXC1. Conclusions: The findings revealed that LINC01614 contributes to the progression of OSCC by targeting the FOXC1 signaling pathway. The study provides insights into a novel mechanistic process to regulate the development of OSCC, and established a possible target for the therapeutic management of OSCC.

Tissue source may affect the esophageal flora in patients with esophageal squamous cell carcinoma

Tissue source may affect the esophageal flora in patients with esophageal squamous cell carcinoma

Abstract B028: Analytical validation of tumor-informed whole genome sequencing analyses for detection of molecular residual disease in solid tumors

Abstract Introduction: Across all patients diagnosed with solid tumors, approximately two-thirds initially present with locoregional disease and are potentially eligible for curative-intent intervention. However, there is a significant subset of patients for which residual tumor cells remain afterwards, potentially leading to disease recurrence. It has been reported that these residual tumor cells, representative of molecular residual disease (MRD), can be detected through analyses of circulating tumor DNA (ctDNA), often earlier than can be achieved through radiographic imaging or clinical presentation. Methods: Labcorp Plasma Detect was utilized to analyze patient-matched tumor, germline (white blood cells), and cell-free or contrived DNA samples through whole genome sequencing at approximately 80x, 40x, and 30x depth, respectively. Raw sequencing data were demultiplexed and trimmed for read quality, then aligned to the hg19 human reference genome. Tumor-specific single nucleotide variants were identified from the tumor and germline datasets and used to determine the presence of cancer within cell-free or contrived DNA samples through a random forest machine learning model. ctDNA status was determined based on the level of signal compared to a reference cohort of noncancerous donor plasma samples (n=80). Results: Analytical specificity was determined through analysis of five noncancerous donor plasma samples with five replicates each, evaluated against 60 somatic mutation profiles obtained from patients with stage III colon cancer, demonstrating a specificity of 99.4% (835/840). Analytical sensitivity was assessed using three contrived cell line models from breast and melanoma tumors, analyzed across three tumor content levels with five replicates at each level and resulted in a limit of detection of 0.005% tumor content using a 100% hit-rate model. Precision, repeatability, and reproducibility were demonstrated within and across runs, operators, and instruments, resulting in a 100% detection rate at 0.05% tumor content with a coefficient of variation of 8.5%. Finally, analytical accuracy compared to an independent tumor-informed ctDNA MRD approach was determined through analysis of 39 clinical cases obtained from patients with bladder, breast, colorectal, and lung cancer, yielding a positive percent agreement of 91% (10/11) and negative percent agreement of 100% (24/24). The prognostic value of ctDNA MRD was demonstrated in stage III colon cancer and clinical validation results for HPV-negative head and neck squamous cell carcinoma will be presented separately. Conclusions: There is significant potential for tumor-informed, non- bespoke MRD approaches for ctDNA detection across a broad range of solid tumor types and curative-intent clinical settings. These data demonstrate the analytical performance of Labcorp Plasma Detect in a CAP/CLIA laboratory for pan-solid tumor research use together with investigational use in early stage colon cancer to support the interventional MEDOCC-CrEATE clinical trial. Citation Format: Kaitlin Victor, Andrew Georgiadis, Ellen Verner, Antoine Simmons, David Riley, James R White, Christopher Greco, Liam Cox, Jesse Fox, Jennifer B Jackson, Eric A Severson, Brian J Caveney, Marcia Eisenberg, Taylor J Jensen, Shakti Ramkissoon, Samuel V Angiuoli, Amy Greer, Kenneth Valkenburg, Mark Sausen. Analytical validation of tumor-informed whole genome sequencing analyses for detection of molecular residual disease in solid tumors [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B028.

Abstract B031: Immune cell composition in surgical drain fluid as a novel prognostic marker in head and neck squamous cell carcinoma

Abstract Background: Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy globally, and the vast majority of cases are treated with primary surgical resection and adjuvant therapy. Our group has pioneered the use of HNSCC circulating tumor DNA (ctDNA) in surgical effluent collected postoperatively from surgical drains (surgical drain fluid, SDF) as a novel proximal bioanalyte for measuring minimal residual disease after surgery. We now profile the immune cell composition of SDF compared to paired peripheral blood mononuclear cells (PBMCs), and correlate SDF immune cell profiles with clinical variables and oncologic outcomes in HNSCC. Method: SDF and peripheral venous blood were collected 24 hours postoperatively from 58 HNSCC patients, including 11 HPV (+) and 47 HPV (-) patients undergoing standard of care surgical resection, lymph node dissection, and adjuvant therapy. Freshly collected SDF cells and PBMCs were cryopreserved until profiling by multicolor flow cytometry for respective leukocyte compositions. Analyses were used to define the following viable CD45+ leukocyte subsets: total T cells, CD4 T-cells CD8 T-cells, Treg cells, B-cells, non- classical monocytes, classical monocytes, natural killer (NK) cells, classical dendritic cells (cDC), plasmacytoid DC (pDC), macrophages, and monocytic myeloid-derived suppressive cells (MDSCs). Immune cell percentages were correlated with clinical characteristics and cancer recurrence. Results: CD45+ leukocytes comprised >99.5% of SDF cells, with very few EPCAM+ tumor cells (< 10^-4). HPV status did not significantly affect SDF immune cell composition, and no significant differences were noted by tumor site (larynx, oral cavity, oropharynx), or extracapsular lymph node extension. Among 13 HPV (-) HNSCC patients with paired peripheral blood drawn concurrently, SDF had a higher proportion of B-cells(16.8% vs. 9%, p=0.044), NK cells(56% vs. 12%, P<0.001), monocytes(14% vs. 1%, P<0.001), and macrophages(3.1% vs. 0.46%, P<0.001) compared to PBMCs, and a lower proportion of CD4+ T cells(1% vs. 24%, P<0.001), CD8+ T cells(0.6% vs. 18%, P<0.001), MDSCs(2.1% vs. 3.4%, P=0.029), and pDCs(0.1% vs. 0.2%, P=0.014). HPV (-) HNSCC patients that went on to recur (n=9) had a significantly higher proportions of SDF monocytes (total, classical, and non- classical) and CD56dimCD16+ NK cells (all P<0.001) compared to non-recurrent patients (n=38). Patients who went on to recur also had a significantly lower proportion of SDF B-cells (P<0.001), CD56bright NK cells(P=0.01), and macrophages (P=0.018) compared to non- recurrent cases. No significant differences were noted with respect to proportions of CD3+, CD4+, CD8+ T-cells, DC, and MDSC cells. Conclusion: SDF from HNSCC patients contains unique immune cell populations and may provide new insights into postoperative tumor immunity. Differences in SDF immune cell composition by oncologic outcome in patients that go on to recur, highlighting potential prognostic and predictive applications of SDF immune cell profiling for adjuvant therapy decision-making. Citation Format: Zhongping Xu, Noah Earland, Lucien Khalil, Lazar Vujanovic, Danny Azmi Elias, Riyue Bao, Jason J. Luke, Aadel A. Chaudhuri, Jose P. Zevallos. Immune cell composition in surgical drain fluid as a novel prognostic marker in head and neck squamous cell carcinoma [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B031.

Abstract A002: The clinical relevance of circulating tumor cell size in head and neck squamous cell carcinoma patients

Abstract This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR002) of the Conference Program/Proceedings. Citation Format: Hyeongjung Woo, Hyun Young Shin, So Yeon Oh, Sun Min Lee, Minseok S. Kim. The clinical relevance of circulating tumor cell size in head and neck squamous cell carcinoma patients [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A002.

Abstract A019: Analytical and clinical validation of HPV-SEQ, an NGS-based liquid biopsy platform for detection and quantification of human papilloma virus circulating tumor DNA

Abstract Human papillomavirus (HPV) infection is the primary causative factor driving the rising incidence of oropharyngeal squamous cell carcinoma (OPSCC). High-risk HPV16 and HPV18 strains are responsible for the majority of HPV related OPSCC cases, with ∼90% and 3-5% respectively. As HPV positivity is associated with improved treatment response and prognosis, correct determination of HPV status is critical for risk stratification and treatment management. While diagnostic tumor biopsy remains a useful tool for HPV profiling, the drawbacks of this approach include pain and discomfort caused by its invasive nature, as well as inability to repeat multiple serial biopsies over time. Noninvasive detection of circulating tumor (ct)HPV-DNA in plasma has emerged as an attractive investigative strategy for early disease diagnosis, real-time response monitoring, and cancer surveillance. However, to date, there is no standard or routine liquid biopsy-based HPV testing for patients with OPSCC. The accuracy of HPV detection in plasma depends on the metrics of the specific method used. Comparison of the most common approaches for detection of ctHPV-DNA shows that next-generation sequencing (NGS)-based assays are superior to that of qPCR and ddPCR, especially in patients with low disease burden. Furthermore, as the majority of current liquid biopsy tests are designed to detect viral E6 and E7 genes, it may comprise a potential challenge for detection of patient-derived ctHPV-DNA among individuals treated with therapeutic vaccines expressing non-oncogenic E6/E7 constructs, actively evaluated in clinical trial settings. To this end, we have developed HPV-SEQ, a novel NGS method designed to achieve maximum sensitivity for HPV16/18 L1 gene in DNA libraries prepared from plasma of OPSCC patients. Here we describe the thorough analytical characterization and clinical validation studies of this lab-developed test conducted and certified under clinical laboratory improvement amendments regulations. HPV-SEQ assay were assessed using both contrived samples and biological specimens by measuring a set of performance attributes including limits of blank, limits of detection, limits of quantitation, accuracy and precision. The assay has demonstrated a robust quantitative detection across a wide ctHPV-DNA range, with low level of background signal (<0.05 copies/sample) in healthy donors and qualitative cut-off of >1.39 copies for both HPV strains (ensuring sufficient differentiation between negative and low positive samples), as well as outstanding sensitivity and specificity, approaching 100%. Given the rapid decrease in sequencing cost and effective integration of sequencing data in clinical care, NGS is likely to displace other HPV detection tests over the next few years. As such, our well-validated, ultra-sensitive, plasma-based HPV profiling method with optimal analytical performance may represent a significant advancement in risk stratification, treatment management, and surveillance for patients with OCSCC and other HPV- driven malignancies. Citation Format: Samaneh Eickelschulte, Anna Starus, David H Murray, Anja K Keyser, Oliver Schauer, Johannes Fredebohm, Frederick Jones, Alexander T Pearson, Everett E Vokes, Ari J Rosenberg, Nishant Agrawal, Evgeny Izumchenko. Analytical and clinical validation of HPV-SEQ, an NGS-based liquid biopsy platform for detection and quantification of human papilloma virus circulating tumor DNA [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A019.

Associations of radiotherapy receipt with lung cancer risk by histological subtype among breast cancer survivors in the United States

AbstractRadiotherapy for breast cancer has been associated with an increased risk of secondary malignancies, including primary lung cancer. Whether this association varies by histological subtype of lung cancer remains unknown. Based on the data from 12 Surveillance, Epidemiology, and End Results registries, we examined the association between radiotherapy receipt and the risk of subtype‐specific subsequent primary lung cancer (SPLC) among female first primary breast cancer cases diagnosed between ages 20 and 84 from 1992 to 2020. More than half (53%) of the 550,007 breast cancer survivors identified had undergone radiotherapy as part of their initial breast cancer treatment. Over an average follow‐up of 9.7 years, 8014 survivors developed SPLCs. For small‐cell carcinoma, the standardized incidence ratio (SIR) compared with the general population was higher for survivors who received radiotherapy (SIR = 1.15, 95% confidence interval [CI] = 1.06–1.25) but similar for those who did not receive radiotherapy (SIR = 1.00, 95% CI = 0.91–1.09), with the difference in SIRs being statistically significant (p = .003). Similar associations were found for squamous cell carcinoma (SIRyes = 1.16, 95% CI = 1.08–1.24 vs. SIRno/unknown = 1.06, 95% CI = 0.98–1.15; p = .07). The increased risks were confined to ipsilateral SPLC, with the greatest SIRs for small‐cell carcinoma occurring 5–10 years since breast cancer diagnosis (SIR = 1.83, 95% CI = 1.53–2.19) and for squamous cell carcinoma with a latency of 10 years or more (SIR = 1.64, 95% CI = 1.42–1.88). In contrast, the risk of developing adenocarcinoma did not vary by radiotherapy receipt (SIRyes = 1.23, 95% CI = 1.18–1.28 vs. SIRno/unknown = 1.17, 95% CI = 1.12–1.22; p = .18), indicating additional risk factors in play. The findings suggest a distinct carcinogenic pathway of radiation‐induced lung cancer across histological subtypes and may inform risk‐stratified surveillance guidelines for SPLC.

Abstract A043: Tumor-informed whole genome sequencing of ctDNA from lymph and plasma detects molecular residual disease in HPV-negative head and neck cancer patients

Abstract Introduction: Recurrence after tumor resection is a major cause of failure in HPV-negative head and neck squamous cell carcinoma (HNSCC), with up to 50% of patients recurring within 2 years. There is unmet need for an accurate diagnostic test that can predict risk of recurrence prior to adjuvant therapy selection. We previously demonstrated that ctDNA in lymphatic exudate collected via surgical drains (“lymph”) accurately identifies molecular residual disease (MRD) and outperforms plasma in HNSCC patients using a targeted sequencing (TS) approach. While TS assays are robust and sensitive, they have key limitations, namely limited ctDNA features, higher cost, longer turnaround times, and lower throughput. Here we evaluate whole genome sequencing (WGS) for detection of MRD in both lymph and plasma sampled 24 hours after surgery. Methods: Lymph, plasma, and whole blood were collected 24 hours postoperatively from 30 HPV-negative HNSCC patients (stages I – IV) along with resected tumor. To detect ctDNA, we utilized WGS analyses of patient matched tumor, germline (blood), and cfDNA from lymph or plasma samples at approximately 150x, 30x, and 30x depth, respectively. Tumor- specific single nucleotide variants were identified from tumor and germline datasets, from which a candidate variant set was used to determine the presence of ctDNA within lymph or plasma DNA through a random forest machine learning model. ctDNA status was determined based on the level of signal compared to a reference population of noncancerous donor plasma samples (n=80). 5 tumors and 1 lymph sample failed QC; in total, 24 patients remained, yielding 10 patients with disease recurrence and 14 with no evidence of disease with >1 year of follow-up. The Kaplan-Meier (KM) estimator with log-rank test and Cox proportional-hazards model were used for survival analyses. Results: KM survival analyses indicated that WGS of lymph ctDNA accurately predicts recurrence (sensitivity (SN) = 50%, specificity (SP) = 93%; p = 0.003, Hazard ratio (HR) = 6.0), consistent with previously published results using a TS MRD approach in this cohort (SN = 80%, SP = 64%, p = 0.02, HR = 5.3). WGS of lymph ctDNA outperformed plasma at this timepoint (SN = 30%, SP = 93%, p = 0.03, HR = 4.2). WGS of lymph ctDNA also showed enhanced performance in locoregional relapse (n=21, SN = 57%, SP = 93%; p = 0.006, HR = 6.7). Finally, the ctDNA fraction of proximal lymph was higher than paired plasma in all locoregional recurrences (n=7), while peripheral plasma had a higher ctDNA fraction in 2 out of 3 distant recurrences. Conclusions: Tumor-informed WGS analysis of lymph ctDNA represents a robust MRD approach in HPV-negative HNSCC. Lymph outperforms time-matched plasma for prediction of recurrence, particularly in patients with locoregional relapse. Accurate MRD identification in the immediate post-surgical timeframe suggests that lymph analysis has potential to augment pathology and longitudinal plasma testing to provide more personalized adjuvant treatment decision-making in patients with HPV-negative HNSCC. Citation Format: Zhuosheng Gu, Seka Lazare, Noah Earland, Marra S Francis, Adam Harmon, Megan Long, Amy Greer, Ellen Verner, Andrew Georgiadis, Mark Sausen, Shakti Ramkissoon, Jose P Zevallos, Aadel A Chaudhuri, Taylor J Jensen, Wendy Winckler. Tumor-informed whole genome sequencing of ctDNA from lymph and plasma detects molecular residual disease in HPV-negative head and neck cancer patients [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A043.

Quantitative proteogenomic characterization in MUC1 and MUC4 in oral squamous cell carcinoma, oral potentially malignant disorders, and normal oral mucosa in carcinogenesis

Quantitative proteogenomic characterization in MUC1 and MUC4 in oral squamous cell carcinoma, oral potentially malignant disorders, and normal oral mucosa in carcinogenesis

Primary Cutaneous Rhabdoid Squamous Cell Carcinoma: A Case Report and Review of Molecular Features

Primary Cutaneous Rhabdoid Squamous Cell Carcinoma: A Case Report and Review of Molecular Features

Risks of malignancies related to disease-modifying antirheumatic drugs in rheumatoid arthritis: a pharmacovigilance analysis using the FAERS database

ObjectivesOver the years when disease-modifying antirheumatic drugs (DMARDs) have been used in rheumatoid arthritis patients, reports of malignancies have emerged. This study aims to investigate the association between malignancies and DMARDs by using data extracted from the Food and Drug Administration Adverse Event Reporting System (FAERS).MethodsFAERS data (January 2019 to December 2023) were reviewed. For each drug-event pair, the disproportionality analysis was conducted to evaluate the risk of malignancy. Multivariate logistic regression was implemented to mitigate potential biases. Moreover, the time to onset of malignancy was also evaluated.ResultsWe conducted a detailed search for rheumatoid arthritis indications and identified a total of 17,412 adverse event reports associated with malignancies, with selective DMARDs designated as the role code “primary suspect”. At the preferred term level, there were 198 positive signals, among which the lower limit of the 95% confidence interval for the information component is 3.55 for squamous cell carcinoma of the skin, 2.39 for breast cancer, and 2.27 for lymphoproliferative disorder. In comparison to other DMARDs, targeted synthetic DMARDs were associated with a broader range of malignancies at both preferred term and Standardized MedDRA Queries levels. The number of adverse events reported in female patients is approximately 2–3 times higher than men, and the median age across the population was approximately 62 years. In terms of onset time, the conventional synthetic DMRADs exhibited a relatively longer median time, ranging from 3.58 to 7.08 years, while the targeted synthetic DMARDs demonstrated a shorter median time of 0.83–1.67 years.ConclusionOur study uncovers varying degrees of malignancy risks related to DMARDs, with a significantly higher risk observed in targeted synthetic DMARDs. Additionally, novel malignancy signals, not documented in product labels, have been detected. In the future, further research will be necessary to validate our findings.

Abstract PR002: The clinical relevance of circulating tumor cell size in head and neck squamous cell carcinoma patients

Abstract Circulating tumor cells (CTCs) play a critical role in the metastatic process of cancer and are widely recognized as important biomarkers that provide valuable information related to tumor characteristics. While most research to date has focused on the biological markers and molecular properties of CTCs, their physical characteristics, particularly size, have been relatively underexplored. However, recent studies suggest that smaller CTCs exhibit enhanced tumor- initiating and metastatic abilities compared to larger CTCs. This indicates that CTC size is not merely a physical trait but may directly influence the invasiveness and metastatic potential of tumors. Such findings suggest that CTC size could be an important indicator for predicting tumor malignancy and provide a new perspective on understanding cancer progression and metastasis. As such, there is a growing emphasis on research that considers the size of CTCs. In this study, it was speculated whether CTC size holds clinical significance in patients with head and neck squamous cell carcinoma (HNSCC). The study was involved with 20 HNSCC patients, analyzing the morphological characteristics of CTCs collected from blood samples before and after treatment, while exploring the potential for monitoring treatment response. Our results found that CTCs were detected 20% more frequently in oropharyngeal cancer (OPC) patients compared to non-OPC patients. This is likely due to the rich vascularity in the oropharyngeal region, which may facilitate greater entry of CTCs into the bloodstream. Additionally, the cell size (23.9 μm in average) and nuclear size (16.4 μm in average) of HPV-positive patients were 32% and 10% larger, respectively, than those of HPV-negative patients (cell size: 18.1 μm; nuclear size: 12.60 μm). This suggests that the sizes of CTC and their nuclei may be associated with HPV status, which is presumed to be due to HPV causing disruptions in the cell cycle. In addition, we could successfully cultivate the CTCs obtained from HPV-positive patients with relatively smaller CTCs for 30 days, predicting that small CTCs have affordable for CTC culture. In conclusion, this study confirms that the physical characteristics of CTCs, particularly size, are linked to HPV infection status in HNSCC patients. CTC- and nucleus- size are closely related to tumor characteristics, demonstrating the potential for developing diagnostic and prognostic tools based on these physical traits. Although more in-depth research is required, poor prognosis in HPV- positive patients may also be related to the physical characteristics of CTCs. Citation Format: Hyeongjung Woo, Hyun Young Shin, So Yeon Oh, Sun Min Lee, Minseok S. Kim. The clinical relevance of circulating tumor cell size in head and neck squamous cell carcinoma patients [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr PR002.

Management of Carcinoma of Esophagus in Old Patient with Panchagavya Therapy as a Palliative Care-A Case Report

ABSTRACT Esophageal cancer is a male-dominant, aggressive malignancy with a five-year survival rate of 15%-20%, predominantly affecting developed nations. Esophageal squamous cell carcinoma (ESCC) is the most common subtype. This case study involves a 70-year-old man with ESCC, who was suffering from difficulty swallowing, inability to burp, constant nausea, and occasional trouble swallowing water for two months. He was successfully treated with Panchagavya Chikitsa, which included Panchagavya Ghrita, Gomutra Arka, Kwath of Amla-Yashtimadhu-Shatpushpa, Haritaki Churna, Haridra Ghanvati, and Vibhitaki Avaleha, along with Nasya, Nabhipurana with Panchagavya Ghrita, and Matra Vasti with Tila Taila over six months. Significant symptom improvement was observed, attributed to the Rasayana (rejuvenative), Balya (strengthening), and Jeevaniya (vitality-enhancing) properties of the treatments. These properties inhibit cancer growth, nourish tissues, and restore balance due to their antioxidant effects. Panchagavya formulations showed promising results without adverse effects, highlighting their efficacy in improving the quality of life for this geriatric patient with ESCC.

Recurrence-Free Survival Prediction for Anal Squamous Cell Carcinoma After Chemoradiotherapy using Planning CT-based Radiomics Model

Abstract Objectives Approximately 30% of non-metastatic anal squamous cell carcinoma (ASCC) patients will experience recurrence after chemoradiotherapy (CRT), and currently available clinical variables are poor predictors of treatment response. We aimed to develop a model leveraging information extracted from radiation pretreatment planning CT to predict recurrence-free survival (RFS) in ASCC patients after CRT. Methods Radiomics features were extracted from planning CT images of 96 ASCC patients. Following pre-feature selection, the optimal feature set was selected via step-forward feature selection with a multivariate Cox proportional hazard model. The RFS prediction was generated from a radiomics-clinical combined model based on an optimal feature set with five repeats of nested five-fold cross validation. The risk stratification ability of the proposed model was evaluated with Kaplan-Meier analysis. Results Shape- and texture-based radiomics features significantly predicted RFS. Compared to a clinical-only model, radiomics-clinical combined model achieves better performance in the testing cohort with higher C-index (0.80 vs 0.73) and AUC (0.84 vs 0.78 for 1-year RFS, 0.84 vs 0.79 for 2-year RFS, and 0.85 vs 0.81 for 3-year RFS), leading to distinctive high- and low-risk of recurrence groups (p < 0.001). Conclusions A treatment planning CT based radiomics and clinical combined model had improved prognostic performance in predicting RFS for ASCC patients treated with CRT as compared to a model using clinical features only. Advances in knowledge The use of radiomics from planning CT is promising in assisting in personalized management in ASCC. The study outcomes support the role of planning CT-based radiomics as potential imaging biomarker.

Abstract B048: Microfluidic isolation of immune cell-derived extracellular vesicles in head and neck cancer patients on immune-checkpoint blockade

Abstract Background: Immune-checkpoint blockade (ICB), with or without chemotherapy, is the standard of care for recurrent/metastatic head and neck squamous cell carcinoma (HNSCC). However, only a minority of patients get clinical benefit from it. The combined expression of PD-L1 in a tumor section is currently the only biomarker approved for response prediction. Yet, its invasive nature and suboptimal predictive performance underscores the need for new, non- invasive surrogates of immune activity in patients undergoing ICB. Extracellular vesicles (EVs) carry key molecules critical for cell-cell communication and have the potential to be used as a non-invasive, real-time biomarker of systemic immune activity, essential for success in ICB- based therapy. Methods: We used a multichannel, microfluidic device that employs an immune- affinity approach, ‘herringbone chip’ (EVHB-Chip), to retrospectively isolate EVs from twelve HNSCC patients before and after the first cycle of ICB. One mL of plasma was used per patient per time point. For each patient, two devices were serially used, with each sample continuously following through each chip in a non-destructive manner—the first device aimed to isolate B cell EVs (antibodies against CD19 and CD20). The second device was optimized to capture T cell EVs (against CD274/PD-L1). Digital-droplet PCR was performed after EVs were isolated to detect the RNA transcripts in the EV cargo. Serial sampling was performed for six out of the twelve patients. Response assessment was conducted via RECIST1.1 criteria or clinical evaluation. Results: Most of our cohort was male 7/12 (57%) and had oral cavity 4/12 (33%) or oropharynx 4/12 (33%) tumors. Additionally, 11/12 (91%) patients received ICB monotherapy. Most of the cohort had a PD-L1 ≤ 1 (7/12; 58%), 7/12 patients experienced disease progression (PD), 4/12 experienced stable disease (SD), and one patient had a partial response (PR). Most of the RNA signal in pre-ICB samples was obtained from CD20 (12/12; 100%), followed by CD45 (10/12; 83%), CD19 (9/12; 75%), and CD274/PD-L1 (5/12; 41%). No difference was found in the RNA signal pre-ICB between responders and non-responders (p=0.1). For the patients with serial sampling available, the signal from CD274/PD-L1 increased after one infusion of ICB in 4/6 patients and remained negative in one. A trend towards a high amount of T cell-derived EVs was observed for the entire cohort (p=0.07; p=0.08) when comparing the amount of RNA via our EV marker (GAPDH, ACTB) between T and B cell devices. Patients with higher than 11.4/mL copies of CD19 (median value for the cohort) in the pre-ICB plasma sample tended to have a higher overall survival (HR 0.26, 95% CI: 0.06-1.13; p-value: 0.05) and progression-free survival (HR 0.28, 95% CI: 0.07-1.16; p-value: 0.06). Conclusion: The EVHB-Chip allowed for the detection and profiling of T and B cell-derived EVs in pre-ICB plasma samples of HNSCC patients undergoing ICB in this pilot cohort. Higher CD19 signal pre-ICB could potentially impact response and survival outcomes in ICB-based therapies. Citation Format: Andrew R. Levy, Daniel A Ruiz Torres, Uma Paithankar, Raheel Ahmad, Daniel C Rabe, Daniel L Faden, Shannon L Stott. Microfluidic isolation of immune cell-derived extracellular vesicles in head and neck cancer patients on immune-checkpoint blockade [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B048.

External Beam Radiotherapy for Malignant Central Airway Obstruction in a Remote Rural Patient: A Case Study

Introduction: Malignant central airway obstruction (MCAO) is a challenging therapeutic scenario, caused by tumour burden which limits airflow within the large airways. Squamous cell carcinoma (SCC) of the lung accounts for 50% of MCAO seen in the setting of non-small cell lung cancer. Here, we present the challenging case of a 63-year-old Indigenous Australian Female rom a remote rural community with background history of metastatic SCC of presumed pulmonary origin, who presented with a 1-week history of dyspnoea, stridor, and hoarse voice. Case Presentation: A CT chest and neck demonstrated a superior mediastinal mass compressing the trachea to a narrowest diameter of 3 mm. The patient was stabilised in her local health centre, then transferred to our tertiary care facility for further evaluation and management. This stenotic lesion was not amenable to bronchoscopic or surgical intervention but was instead treated successfully with a short course of external beam radiotherapy, to total dose of 25Gy delivered over 10 fractions. The patient had almost complete resolution of respiratory symptoms with no significant radiotherapy related toxicity. Conclusion: Here, we discuss a challenging case of MCAO in a remote rural Australian patient and demonstrate the utility of external beam radiotherapy in this setting. We also discuss aspects of cancer related healthcare disparities which exist for Indigenous Australians. In doing so, we wish to highlight the need for improved cancer healthcare for such communities.

Cytokines levels in saliva in periodontitis, oral potentially malignant disorders and squamous cell carcinoma

Biopsy and histopathological examination continue to be the gold standards for diagnosing oral cancer and oral potentially malignant disorders (OPMD). The prognosis of these conditions is significantly influenced by established factors such as the degree of dysplasia, tumour size, grade, and patient age. While these traditional diagnostic approaches are crucial, there is a growing need for innovative methods to improve diagnosis and monitoring of these conditions. We hypothesize that identifying specific diagnostic biomarkers in human saliva, alongside traditional methods, could enhance clinical outcomes in managing both malignant and non-malignant oral pathologies. In this study, salivary levels of previously described biomarkers BDNF, IL-6, IL-8, IL-10, IL-15, IL-1RA, LIF, and TNF-α were compared among oral cancer patients, patients with OPMD, periodontal disease patients, and healthy volunteers. The results indicated that these cytokines have the potential to distinguish cancer cases, though they are less effective in differentiating between OPMD and periodontitis. Using pairs or groups of biomolecules could improve the ability to differentiate between different pathological entities. We found the following panel IL-6, IL-15, and IL-1RA to be particularly significant as diagnostic biomarkers for oral squamous cell carcinoma and OPMD.

Added-value of dynamic contrast-enhanced MRI to conventional MRI for the differentiation between inflammatory myofibroblastic tumor and squamous cell carcinoma in the sinonasal region

Added-value of dynamic contrast-enhanced MRI to conventional MRI for the differentiation between inflammatory myofibroblastic tumor and squamous cell carcinoma in the sinonasal region

Melanoma Cells from Different Patients Differ in Their Sensitivity to Alpha Radiation-Mediated Killing, Sensitivity Which Correlates with Cell Nuclei Area and Double Strand Breaks

Background/Objective: In this study, for the first time, we examined and compared the sensitivity of four patient-derived cutaneous melanoma cell lines to alpha radiation in vitro and analyzed it in view of cell nucleus area and the formation of double-strand breaks (DSB). Melanoma cells sensitivity to alpha radiation was compared to photon radiation effects. Furthermore, we compared the sensitivity of the melanoma cells to squamous cell carcinoma. Methods: Human melanoma cell lines YDFR.C, DP.C, M12.C, and M16.C, and the squamous cell carcinoma cell line, CAL 27, were irradiated in vitro using Americium-241 as alpha-particle source. Cells were irradiated with doses of 0 to 2.8 gray (Gy). Cell viability, DNA DSB, and nuclear size were measured. Results: 1. Alpha radiation caused death and proliferation arrest of all four melanoma cell lines, but inter-tumor heterogeneity was observed. 2. The most sensitive cell line (DP.C) had a significantly larger nucleus area (408 µm2) and the highest mean number of DSB per cell (9.61) compared to more resistant cells. 3. The most resistant cell, M16.C, had a much lower nucleus area (236.99 µm2) and DSB per cell (6.9). 4. Alpha radiation was more lethal than photon radiation for all melanoma cells. 5. The SCC cell, CAL 27, was more sensitive to alpha radiation than all melanoma cells but had a similar number of DSB (6.67) and nucleus size (175.49 µm2) as the more resistant cells. 6. The cytotoxic effect of alpha radiation was not affected by proliferation arrest after serum starvation. 7. Killing of cells by alpha radiation was marginally elevated by ATR or topoisomerase 1 inhibition. Conclusions: This study demonstrates that various human melanoma cells can be killed by alpha radiation but exhibit variance in sensitivity to alpha radiation. Alpha radiation applied using the Intra-tumoral Diffusing alpha-emitters Radiation Therapy (Alpha DaRT) methodology may serve as an efficient treatment for human melanoma.