Head Squamous Cell Research Articles

577. OUTCOMES OF SIMULTANEOUS RESECTION AND RECONSTRUCTION OF ESOPHAGEAL SQUAMOUS CELL CARCINOMA SYNCHRONOUSLY ASSOCIATED WITH HEAD AND NECK CANCER

Abstract In 1953, a study by Slaughter et al. on oral cancer proposed that field cancerization is the phenomenon of widespread carcinogenesis across multiple regions due to long-term exposure to common cancer-causing factors. Esophageal squamous cell carcinoma (ESCC) and head and neck cancer (HNC) share common risks, such as smoking and alcohol consumption, and are often observed synchronously. Therefore, treatment strategies based on tumor stages are important. In our hospital, for synchronous HNC and thoracic ESCC, local resection is indicated if the HNC is ≤T2, and resection with reconstruction is indicated if the HNC is ≥T3, and a one-stage operation for thoracic ESCC and HNC was generally performed. Here, we examined treatment strategies based on the results of simultaneous resection and reconstruction of synchronous thoracic ESCC and HNC in our hospital. A total of 174 patients with esophageal cancer underwent surgery at our hospital between January 2008 and December 2023. Among them, 16 (9%) had 18 lesions of synchronous HNC and nine patients (5%) underwent simultaneous resection and reconstruction. Patients with thoracic ESCC had Ut/Mt/Lt; 1/6/2; T-factor, T1/2/3; 5/2/2; N-factor, N0/1:6/3; and clinical stage, stage I/II; 2/6. For synchronous HNC, hypopharyngeal/oropharyngeal/laryngeal/cervical esophageal cancer; 5/2/1/2; T factor was T2/3/4; 1/6/3; N factor was N0/1/2c/3a/3b; 4/2/2/2/1/1; and clinical stage was stage II/III/IV; 1/5/4. Six patients underwent surgery after preoperative chemotherapy or chemoradiotherapy. Total pharyngo-laryngo-esophagectomy (TPLE) was performed using a thoracoscopic transthoracic approach in six patients, transhiatal approach in two, and mediastinoscopic approach in one. Regarding reconstruction, a gastric tube was used in nine patients, including eight patients in whom it was interposed with the free jejunal autograft. The median operative time was 929 min (590–1248 min) and median blood loss was 622 mL (213–1764 mL). Only patients with skin valve necrosis underwent reoperation; however, there were no hospital deaths, and the median postoperative hospital stay was 39 days (28–59 days). Recurrence was observed in four cases. All cases were considered to be due to HNC, and three were stage IV cases with advanced lymph node metastasis. Simultaneous resection and reconstruction for synchronous HNC and the thoracic esophagus is a highly invasive treatment that can be performed safely. Recurrence has been observed in advanced cases of HNC, and further development of multidisciplinary treatment, including preoperative chemotherapy, is necessary.

Current treatments for oropharyngeal squamous cell carcinoma and the move towards molecular therapy.

In this review, we discuss oropharyngeal squamous cell carcinoma (OPSCC) treatment options with a focus on the molecular mechanisms of OPSCC in head and neck squamous cell carcinoma (HNSCC) and head and neck cancers (HNCs). Treatment can be radical intent (aim for cure) or palliative intent (aim for disease control and symptom management). OPSCC is a prominent subset of HNSCCs in Australia and the Western World. We looked at the current conventional treatment options with an overview of recent advances and future endeavours. We identified that radiotherapy is the primary management for OPSCC in most countries, including the USA, UK, NZ, and Australia. In contrast, surgery is only considered for superficial OPSCC or neck surgery. If surgery is incomplete, then definitive management still requires radiotherapy. Molecular therapy is largely at the preclinical stage, with cetuximab, nivolumab, pembrolizumab, Lenvatinib, and bevacizumab being tested clinically currently.

The potential of hydrogel-free tumoroids in head and neck squamous cell carcinoma.

Head and neck malignancy, and in particular squamous cell carcinoma (SCC), is responsible for a significant disease burden globally. The lack of an optimal invitro model system to accurately recapitulate invivo response to therapy in HNSCC remains a challenge. The development of patient-derived three-dimensional tumour cultures, or tumoroids, has enabled improved modelling of the tumour microenvironment through simulation of important characteristics such as tumour hypoxia, cell-cell interactions and nutrient diffusion characteristics. We performed a comprehensive English-language literature review of current methods of tumoroid development utilising Matrigel and Cultrex Basement Membrane Extract 2 (key terms: tumour organoids, tumoroids, hydrogels, Matrigel, Cultrex, squamous cell carcinoma, head and neck)-two common proprietary murine-derived hydrogels containing extracellular matrix proteins. Nascent literature on the establishment of a novel hydrogel-free platform for tumoroid development as distinct from these existing methods was also explored. Whilst useful for facilitating cell-matrix interactions and providing a scaffold for three-dimensional cell growth and organisation, murine-derived cell matrix methods were noted to have notable limitations including temperature sensitivity and the medium forming a barrier to analysis of the supernatant. A novel hydrogel-free method of establishing invitro tumoroid cultures has been subject to experimentation in colorectal but not head and neck malignancy. The absence of a hydrogel provides for the de novo synthesis of extracellular matrix native to the tumour and self-organisation of cells within this scaffold through the use of ultralow attachment plates. This model demonstrates similar structural and physiological properties to native tissue, whilst enabling more accurate biomimicry of the tumour microenvironment for drug testing. In the absence of prior experimentation on a hydrogel-free method for establishing HNSCC tumoroids, and comparisons between hydrogel and hydrogel-free models, the future development of a comparative protocol encompassing recruitment, collection, processing and analysis represents a valuable opportunity.

The SRC family kinase inhibitor NXP900 demonstrates potent antitumor activity in squamous cell carcinomas

NXP900 is a selective and potent SRC family kinase (SFK) inhibitor, currently being dosed in a phase 1 clinical trial, that locks SRC in the “closed” conformation, thereby inhibiting both kinase-dependent catalytic activity and kinase-independent functions. In contrast, several multi-targeted kinase inhibitors that inhibit SRC, including dasatinib and bosutinib, bind their target in the active “open” conformation, allowing SRC and other SFKs to act as a scaffold to promote tumorigenesis through non-catalytic functions. NXP900 exhibits a unique target selectivity profile with sub-nanomolar activity against SFK members over other kinases. This results in highly potent and specific SFK pathway inhibition. Here, we demonstrate that esophageal squamous cell carcinomas (ESCC) and head and neck squamous cell carcinomas (HNSCC) are exquisitely sensitive to NXP900 treatment in cell culture and in vivo, and we identify a patient population that could benefit from treatment with NXP900.

Mutant Nrf2E79Q enhances the promotion and progression of a subset of oncogenic Ras keratinocytes and skin tumors

Squamous cell carcinomas (SCCs), including lung, head & neck, bladder, and skin SCCs often display constitutive activation of the KEAP1-NRF2 pathway. Constitutive activation is achieved through multiple mechanisms, including activating mutations in NFE2L2 (NRF2). To determine the functional consequences of Nrf2 activation on skin SCC development, we assessed the effects of mutant Nrf2E79Q expression, one of the most common activating mutations in human SCCs, on tumor promotion and progression in the mouse skin multistage carcinogenesis model using a DMBA-initiation/TPA-promotion protocol where the Hras A->T mutation (Q61L) is the canonical driver mutation. Nrf2E79Q expression was temporally and conditionally activated in the epidermis at two stages of tumor development: 1) after DMBA initiation in the epidermis but before cutaneous tumor development and 2) in pre-existing DMBA-initiated/TPA-promoted squamous papillomas. Expression of Nrf2E79Q in the epidermis after DMBA initiation but before tumor occurrence inhibited the development/promotion of 70% of squamous papillomas. However, the remaining papillomas often displayed non-canonical Hras and Kras mutations and enhanced progression to SCCs compared to control mice expressing wildtype Nrf2. Nrf2E79Q expression in pre-existing tumors caused rapid regression of 60% of papillomas. The remaining papillomas displayed the expected canonical Hras A->T mutation (Q61L) and enhanced progression to SCCs. These results demonstrate that mutant Nrf2E79Q enhances the promotion and progression of a subset of skin tumors and alters the frequency and diversity of oncogenic Ras mutations when expressed early after initiation.

A plasma-based proteomic platform for predicting clinical benefit from immune checkpoint inhibitors in multiple cancers.

2568 Background: Immune checkpoint inhibitor (ICI)-based therapies are preferred treatment options for multiple cancer types. However, there is an unmet need for predictive tests that can identify patients likely to benefit. PROphet-NSCLC is a commercially available pretreatment plasma proteomic test that predicts clinical benefit from first-line PD-(L)1 inhibitor-based therapy in patients with metastatic non-small cell lung cancer (NSCLC), guiding the choice between ICI monotherapy versus ICI-chemotherapy. Here, we evaluate the broader clinical utility of PROphet-NSCLC by testing its predictive capabilities across diverse cancer types. Methods: Pre-treatment plasma samples and clinical data were collected for an observational study from patients with metastatic melanoma (n=68) and HPV-related cancers (n=43; NCT03427411) treated with PD-(L)1 inhibitor-based therapies. HPV-related cancers included anogenital squamous cell carcinoma (n=16), cervical carcinoma (n=18), and head and neck squamous cell carcinoma (n=9). Patients were assigned a PROphet-positive or -negative result based on computational analysis of SomaScan-derived proteomic profiles in pre-treatment plasma samples. Overall survival (OS) and progression-free survival (PFS) in PROphet-positive- and -negative groups was analyzed with the Kaplan-Meier method. Hazard ratios (HR) were calculated from univariate and multivariate Cox proportional hazard models. Results: In the melanoma cohort, PROphet-positive patients (n=51) displayed a significant OS benefit in comparison to PROphet-negative patients (n=17; median OS 92.8 months vs. 9.5 months, HR=0.14, 95% CI: 0.06-0.34, p<0.0001), consistent with the clinically validated predictive performance of the test in patients with NSCLC. OS results remained significant after correcting for sex, age, histology, ECOG performance status, and treatment type (HR=0.05, 95% CI: 0.01-0.25, p<0.001). In PROphet-positive patients, PD-1+CTLA-4 inhibitor combination therapy (n=27) was superior to PD-1 inhibitor monotherapy (n=24; OS: median 118.4 vs. 48.3 months, HR=0.58, p=0.24; PFS: median not reached vs. 10.8 months, HR=0.48, p=0.04). PROphet-negative patients displayed similarly poor outcomes with either treatment, providing a rationale to consider alternative therapies for such patients. In the HPV-related cohort, the median OS in PROphet-positive (n=10) vs PROphet-negative (n=33) groups was 43.6 vs 4.4 months (HR=0.22, 95% CI: 0.08-0.59, p=0.001), with similar trends per sub-cohort. Conclusions: Our findings show that the PROphet-NSCLC test can be applied to PD-(L)1 inhibitor-treated melanoma and HPV-related cancers, suggesting applicability to cancers beyond NSCLC. Analysis of additional patient samples is needed to explore the potential utility of PROphet-NSCLC for informing treatment decisions for a broad range of cancer types.

Phase 1b study evaluating the efficacy and safety of ABBV-400, a c-Met–targeting antibody-drug conjugate, in select advanced solid tumor indications.

TPS3162 Background: c-Met (MET protein) is commonly overexpressed in a number of tumors, including hepatocellular carcinoma (HCC), pancreatic ductal adenocarcinoma (PDAC), biliary tract cancers (BTC), esophageal squamous cell carcinoma (ESCC), breast cancer (BC), and head and neck squamous cell carcinoma (HNSCC). Patients with c-Met–overexpressing tumors represent an underserved population, with the need for more effective c-Met–targeting therapies to become available. ABBV-400 is an antibody-drug conjugate, consisting of the c-Met–targeting antibody telisotuzumab conjugated to a potent topoisomerase 1 inhibitor payload. Initial results from the ongoing first-in-human study (NCT05029882) of ABBV-400 in patients with advanced solid tumors indicate a tolerable safety profile, with a maximum tolerated dose of 3 mg/kg once every 3 weeks (Q3W), and promising antitumor activity, with an overall response rate of 24.4% (1). Herein, we describe a signal-seeking study evaluating ABBV-400 treatment in patients with select solid tumors. Methods: Multicenter, open-label, phase 1 signal-seeking study(NCT06084481). Eligible patients (≥18 years) have confirmed locally advanced/metastatic disease measurable per RECIST v1.1 and Eastern Cooperative Oncology Group performance status ≤1. Approximately 220 patients are planned for enrollment across 7 cohorts (HCC, n=40; PDAC, n=40; BTC, n=20; ESCC, n=40; triple-negative BC, n=20; hormone receptor-positive/HER2-negative BC, n=20; HNSCC, n=40). The primary objectives are to assess efficacy and safety/tolerability of ABBV-400 in each tumor indication. Secondary objectives include the evaluation of pharmacokinetics (PK) and immunogenicity of ABBV-400. Pharmacodynamic (PD) and biomarker analyses are exploratory endpoints. Patients receive intravenous ABBV-400 at 3 mg/kg Q3W until disease progression, intolerable toxicity, or any other per-protocol discontinuation criteria. c-Met expression will be assessed retrospectively by immunohistochemistry. The maximum treatment duration is 2 years. Tumor assessments are performed at screening and every 6 weeks from the first dose of study drug, with objective response rate as primary efficacy endpoint and duration of response, clinical benefit rate, progression-free survival, and overall survival as secondary efficacy endpoints. Safety evaluations include adverse events monitoring (graded according to the NCI CTCAE, v5.0), physical examinations, vital sign measurements, ECG variables, and clinical laboratory testing. Blood samples for PK, PD, and biomarker analysis are collected at designated time points throughout the study. Enrollment started in November 2023. As of 19 January 2024, 24 patients have been enrolled. 1. Sharma et al. JCO 2023;41[16 suppl]:3015. Clinical trial information: NCT06084481 .

Abstract CT227: Anti-LAG-3 antibody LBL-007 in combination with anti-PD-1 antibody toripalimab, in patients with advanced malignant tumors: A phase Ib/II, open-label, multicenter, dose escalation/expansion study

Abstract Background: Dual inhibition of PD-1 and LAG-3 may have the potential to increase the immune response against tumor growth synergistically. Here we report the preliminary safety and efficacy results of LBL-007 (an anti-LAG-3 antibody) in combination with toripalimab (an anti-PD-1 antibody) in patients with advanced malignant tumors. Methods: Patients with advanced malignant tumors, including but not limited to nasopharyngeal carcinoma (NPC), non-small cell lung cancer (NSCLC), esophageal squamous cell carcinoma (ESCC), and head and neck squamous carcinoma (HNSCC), with or without prior anti-PD-(L)1 therapy, were enrolled. This trial was divided into two parts: phase Ib (dose escalation study), patients received LBL-007 at 200 mg or 400 mg plus toripalimab at 240 mg (both administered i.v. Q3W), and phase II (dose expansion), patients received LBL-007 at 400 mg and toripalimab at 240 mg (both administered i.v. Q3W). The primary objective was to assess safety and tolerability; the secondary objectives were to assess pharmacokinetics, pharmacodynamics, and preliminary efficacy (per RECIST 1.1). Results: By December 1, 2023, 80 patients were enrolled in phase Ib (10 patients) and phase II (70 patients). Of these, 30 (37.5%) were NPC patients, and 50 (62.5%) had other tumors. The median follow-up was 18.1 months (95% CI: 11.1, NE). No DLTs were observed in phase Ib. Seventy-three patients out of 80 (91.3%) experienced TEAEs of all grades, with 19 (23.8%) having grade ≥3 TEAEs. The most common any grade TEAEs (≥15%) were anaemia (40.0%), hypoalbuminaemia (18.8%), weight decreased (18.8%), hyponatremia (17.5%), proteinuria (17.5%), asthenia (17.5%), lymphocyte count decreased (16.3%), aspartate aminotransferase increased (16.3%), hypertriglyceridaemia (15.0%), and rash (15.0%). Treatment interruption and permanent discontinuation due to TEAEs occurred in 6 (7.5%) patients each. Sixteen patients (20.0%) experienced SAEs. Four patients experienced TEAEs leading to death, all events are related to underlying disease or disease progression, none of which were related to LBL-007 or toripalimab. Out of 75 efficacy evaluable patients, ORR and DCR were 13.3% (10/75) and 48.0% (36/75), respectively. In 29 efficacy evaluable NPC patients, ORR and DCR were 20.7% (6/29), and 62.1% (18/29), respectively. Among the group of 12 anti-PD-(L)1 treatment-naïve NPC patients and 17 anti-PD-(L)1 treated NPC patients, the ORR were 33.3% (4/12) and 11.8% (2/17), DCR were 75.0% (9/12) and 52.9% (9/17), respectively; Of note, the mPFS was up to 11.9 months (95%CI: 1.3, not reached) in anti-PD-(L)1 treatment-naïve NPC patients. Conclusions: LBL-007 in combination with toripalimab is well tolerated in patients with advanced malignant tumors and has demonstrated encouraging preliminary efficacy in patients with advanced NPC with or without prior anti-PD-(L)1 therapy. Clinical trial information: NCT05102006. Citation Format: Yunpeng Yang, Yuanyuan Zhao, Wenfeng Fang, Yan Huang, Yaxiong Zhang, Yi Ba, Zhen Wang, Chao Deng, Desheng Hu, Wei Wang, Guiling Li, Suxia Luo, Zhichao Fu, Haisheng Zhu, Huili Wang, Shiwei Zhao, Tao Li, Charles Cai, Li Zhang. Anti-LAG-3 antibody LBL-007 in combination with anti-PD-1 antibody toripalimab, in patients with advanced malignant tumors: A phase Ib/II, open-label, multicenter, dose escalation/expansion study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT227.

Abstract 1852: NCI’s Clinical Proteomic Tumor Analysis Consortium: A proteogenomic cancer analysis program

Abstract The National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (NCI’s CPTAC) is a national effort to accelerate the understanding of cancer biology through the integration of large-scale proteome and genome analysis, or proteogenomics. The program which is composed of a Tumor Characterization Program and a Translational Research Program (proteomics.cancer.gov) aims to leverage the investments made in cancer research by understanding the molecular changes by proteogenomically characterizing cancers and accelerating the basic discovery toward clinical impact. All data and analytical tools are made broadly available to the research community through public databases to maximize utility and public benefit.CPTAC teams have characterized a plethora of treatment-naïve tumor types, including colorectal, ovarian, breast, clear cell renal cell carcinoma, uterine corpus endometrial carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, brain (including adult, pediatric, and adolescent and young adult), head and neck squamous cell carcinoma, and pancreatic ductal adenocarcinoma (proteomics.cancer.gov/resources/milestones-and-publications). Ongoing projects include acute myeloid leukemia (AML), cutaneous melanoma, sarcoma, oligodendroglioma, gastric, prostate, liver, and thyroid cancers. In the PTRC Program, CPTAC is partnering with NCI-sponsored clinical trials to support clinically relevant research projects that would elucidate biological mechanisms of therapeutic response, resistance, and/or toxicity in cancers including AML, multiple myeloma, melanoma, and lung cancers. All mass spectrometry-based proteomic, genomic, and imaging data (histopathology and radiology) are made publicly available at the CPTAC Proteomic Data Commons (pdc.cancer.gov), Genomic Data Commons (gdc.cancer.gov), and the Cancer Imaging Archive (cancerimagingarchive.net)/Imaging Data Commons (imaging.datacommons.cancer.gov) respectively. CPTAC is also supporting development of new proteogenomic data analysis tools (proteomics.cancer.gov/resources/computational-tools). In addition, the CPTAC Assay Portal (assays.cancer.gov) is a public resource populated with mass spectrometry-based targeted proteomic assays developed by the consortium for quantitatively measuring proteins of interest, including those discovered through comprehensive tumor characterization. Lastly, well-characterized monoclonal antibodies targeting cancer-specific proteins and peptides are also made available at CPTAC’s Antibody Portal (antibodies.cancer.gov). Citation Format: Mehdi Mesri, Eunkyung An, Xu Zhang, Jasmin Bavarva, Ana I. Robles, Tara Hiltke, Henry Rodriguez. NCI’s Clinical Proteomic Tumor Analysis Consortium: A proteogenomic cancer analysis program [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1852.

Abstract PR011: Discovery of VVD-065, a first-in-class allosteric molecular glue of the Keap1-Cul3 E3-ligase complex for the treatment of NRF2-activated cancers

Abstract Somatic gain of function mutations in nuclear factor erythroid 2-related factor 2 (NRF2) transcription factor or loss of function mutations in Kelch-like ECH Associated Protein 1 (KEAP1) E3 ligase, which regulates NRF2 protein levels, are frequently identified in many solid cancers such as non-small cell lung cancer (NSCLC), esophageal squamous cell carcinoma (ESCC), and head and neck squamous cell carcinoma (HNSCC). These genomic alterations drive the activation of cytoprotective NRF2 transcriptional programs. In addition to mutational activation, NRF2 pathway activation has been documented with high frequency in patients without mutations in the pathway. Since tumor cells with hyperactivated NRF2 have been demonstrated to be dependent on this pathway for survival, targeting NRF2 represents an attractive therapeutic approach in tumors with aberrant NRF2 activation. Here, we report the identification of VVD-065, a first-in-class NRF2 inhibitor that acts via an unprecedented mechanism of action. Covalent modification of sensor cysteines on KEAP1 during electrophilic or oxidative stress is known to inhibit KEAP1 mediated NRF2 degradation. In complete contrast, covalent modification of a KEAP1 sensor cysteine by VVD-065 dramatically enhances NRF2 degradation. At low nM concentrations, VVD-065 covalently targets KEAP1 E3 ligase, and allosterically increases the affinity between KEAP1 and CUL3, thereby promoting the formation of active KEAP1-CUL3 E3 ligase complexes and increasing the rate of NRF2 degradation. VVD-065 profoundly reduces NRF2 protein levels in WT KEAP1/NRF2 settings, as well as various KEAP1 and NRF2 mutant settings. Consequently, VVD-065 substantially inhibits NRF2-dependent gene expression and proliferation of NRF2-dependent cell lines. In vivo, oral administration of VVD-065 results in robust degradation of NRF2 and decreased expression of NRF2 target genes. Tumor growth inhibition studies with cell-line derived, and patient derived NSCLC/ESCC xenograft (CDX/PDX) models revealed robust dose-dependent tumor growth inhibition and tumor regression in the absence of any overt toxicity. Since constitutive NRF2 activation often contributes to chemotherapeutic resistance, we also conducted TGI studies with 100+ PDX models to assess combination opportunities with chemotherapeutic agents. A marked combination response was achieved with cisplatin and nab-paclitaxel in various solid tumor types such as NSCLC, ESCC, HNSCC, and uterine cancers. Combination with chemotherapy was safe and well-tolerated in these mouse studies. In summary, we have identified VVD-065, a potent and selective KEAP1 activator that promotes the degradation of NRF2. Degradation of NRF2 led to robust monotherapy response in NRF2-activated cancers and also chemo-sensitized chemo-refractory tumors. A related molecule, acting through the same mechanism of action has now entered into Phase I clinical trials. Citation Format: Nil Roy, Tine Wyseure, I-Chung Lo, Jordon Inloes, Aaron Snead, Steffen Bernard, Justine Metzger, Jonathan Pollock, Stephanie Grabow, Christie Eissler, Melaminah Williams, Sarah Jacinto, Gabe Simon, Todd Kinsella, David Weinstein, Matt Patricelli. Discovery of VVD-065, a first-in-class allosteric molecular glue of the Keap1-Cul3 E3-ligase complex for the treatment of NRF2-activated cancers [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr PR011.

Protocol for the EACH trial: a multicentre phase II study evaluating the safety and antitumour activity of the combination of avelumab, an anti-PD-L1 agent, and cetuximab, as any line treatment for patients with recurrent/metastatic head and neck squamous cell cancer (HNSCC) in the UK

IntroductionHead and neck cancer is the eighth most common cancer in the UK. Current standard of care treatment for patients with recurrent/metastatic squamous cell head and neck carcinoma (HNSCC) is...

Advances in SEMA3F regulation of clinically high-incidence cancers.

Cancer has become a leading cause of morbidity and mortality in recent years. Its high prevalence has had a severe impact on society. Researchers have achieved fruitful results in the causative factors, pathogenesis, treatment strategies, and cancer prevention. Semaphorin 3F (SEMA3F), a member of the signaling family, was initially reported in the literature to inhibit the growth, invasion, and metastasis of cancer cells in lung cancer. Later studies showed it has cancer-inhibiting effects in malignant tumors such as breast, colorectal, ovarian, oral squamous cell carcinoma, melanoma, and head and neck squamous carcinoma. In contrast, recent studies have reported that SEMA3F is expressed more in hepatocellular carcinoma than in normal tissue and promotes metastasis of hepatocellular carcinoma. We chose lung, breast, colorectal, and hepatocellular carcinomas with high clinical prevalence to review the roles and molecular mechanisms of SEMA3F in these four carcinomas. We concluded with an outlook on clinical interventions for patients targeting SEMA3F.

Validating the role of PTGIS gene in colorectal cancer by bioinformatics analysis and in vitro experiments

Prostaglandin I2 synthase (PTGIS) is a member of the cytochrome P450 family. Studies have revealed that differential expression of the PTGIS gene is closely related to the pathological and physiological processes of many diseases, including breast cancer, oral squamous cell carcinoma, and head and neck cancer. However, the mechanism of action of the PTGIS gene in colorectal cancer is not fully understood. This study explored the role of PTGIS in colorectal cancer through comprehensive bioinformatics analysis and in vitro experiments, and found that the expression of PTGIS gene in colorectal cancer tissue was significantly lower than that in normal colorectal tissue (P < 0.05), and high expression of PTGIS gene was associated with poor prognosis in patients (P < 0.05). The KEGG results showed that PTGIS-related genes were mainly enriched in metabolic pathways, arachidonic acid metabolism, steroid biosynthesis, and cancer pathways. The expression of PTGIS may be related to immune infiltration. Cell experiments showed that PTGIS was expressed at a lower level in cancer. Overexpression of PTGIS inhibited apoptosis and promoted proliferation, invasion, and migration ability of SW480 colorectal cancer cells. Analysis of the PTGIS gene in this study provides a theoretical basis for further exploring the pathogenesis of colorectal cancer and finding more accurate new targets for early screening and treatment of the cancer.

Next-Generation Sequencing Analysis of Mutations in Circulating Tumor DNA from the Plasma of Patients with Head-Neck Cancer Undergoing Chemo-Radiotherapy Using a Pan-Cancer Cell-Free Assay.

Using next-generation sequencing (NGS), we investigated DNA mutations in the plasma tumor cell-free circulating DNA (ctDNA) of 38 patients with inoperable squamous cell head neck cancer (SCHNC) before and after the completion of chemoradiotherapy (CRT). Baseline mutations of the TP53 were recorded in 10/38 (26.3%) and persisted in 4/10 patients after CRT. ΤP53 mutations were further detected post CRT in 7/38 additional patients with undetectable mutations at baseline (overall rate 44.7%). Furthermore, 4/38 patients exhibited baseline mutations of the EGFR, AR, FGFR3, and FBXW3, and four new gene mutations were detected after CRT (MTOR, EGFR3, ALK, and SF3B1). Τ4 stage was related with a significantly higher rate of mutations (TP53 and overall). Mutations were observed in 8/30 (26.6%) responders (complete/partial response) vs. in 6/8 (75%) of the rest of the patients (p = 0.03). Significant poorer LRFS was noted for patients with mutations detected before and after CRT (p = 0.02). Patients who had detectable mutations either before or after CRT had significantly worse DMFS (p = 0.04 overall, and p = 0.02 for TP53 mutations). It was concluded that assessment of mutations before and after the end of CRT is essential to characterize patients with a high risk of locoregional recurrence or metastatic progression.

A Random Forest Model for Post-Treatment Survival Prediction in Patients with Non-Squamous Cell Carcinoma of the Head and Neck.

Compared to squamous cell carcinoma, head and neck non-squamous cell carcinoma (HNnSCC) is rarer. Integrated survival prediction tools are lacking. 4458 patients of HNnSCC were collected from the SEER database. The endpoints were overall survivals (OSs) and disease-specific survivals (DSSs) of 3 and 5 years. Cases were stratified-randomly divided into the train & validation (70%) and test cohorts (30%). Tenfold cross validation was used in establishment of the model. The performance was evaluated with the test cohort by the receiver operating characteristic, calibration, and decision curves. The prognostic factors found with multivariate analyses were used to establish the prediction model. The area under the curve (AUC) is 0.866 (95%CI: 0.844-0.888) for 3-year OS, 0.862 (95%CI: 0.842-0.882) for 5-year OS, 0.902 (95%CI: 0.888-0.916) for 3-year DSS, and 0.903 (95%CI: 0.881-0.925) for 5-year DSS. The net benefit of this model is greater than that of the traditional prediction methods. Among predictors, pathology, involved cervical nodes level, and tumor size are found contributing the most variance to the prediction. The model was then deployed online for easy use. The present study incorporated the clinical, pathological, and therapeutic features comprehensively and established a clinically effective survival prediction model for post-treatment HNnSCC patients.

Equivalence of laboratory-developed test and PD-L1 IHC 22C3 pharmDx across all combined positive score indications.

We conducted an analysis across multiple PD-L1 combined positive score (CPS) indications to establish concordance of a 22C3 antibody-based laboratory-developed test (LDT) on the Ventana BenchMark XT or BenchMark ULTRA platform and the regulatory-approved PD-L1 IHC 22C3 pharmDx in cervical cancer (CC), esophageal squamous cell carcinoma (ESCC), head and neck squamous cell carcinoma (HNSCC), triple-negative breast cancer (TNBC), and urothelial carcinoma (UC). Tumor specimens from each tumor type were stained with 22C3 antibody and scored using the 22C3 antibody-based LDT, and scores were compared with those using PD-L1 IHC 22C3 pharmDx. PD-L1 status was measured by the pathologist using CPS as a continuous score and using clinically relevant cutoffs (CC, ≥1 and ≥10; HNSCC, ≥1 and ≥20; ESCC, TNBC, and UC, ≥10). The agreement between the BenchMark platforms and PD-L1 IHC 22C3 pharmDx was assessed by intraclass correlation coefficient (ICC) and a contingency table for clinical interpretation. A total of 522 samples were evaluated for the pan-tumor analysis (CC, n = 77; ESCC, n = 80; HNSCC, n = 126; TNBC, n = 118, UC, n = 121). Most clinical interpretations of PD-L1 status were concordant between the BenchMark XT and PD-L1 IHC 22C3 pharmDx for all five tumor types with regard to negative percentage agreement (NPA; 83-97%), positive percentage agreement (PPA; 86-100%), and overall percentage agreement (OPA; 90-97%); the ICC by tumor type was high (≥0.88). Importantly, the pan-tumor ICC was 0.95 (95% CI 0.94-0.96). Thirty additional TNBC samples were evaluated using the BenchMark ULTRA and PD-L1 IHC 22C3 pharmDx; the NPA, PPA, and OPA were 100%. The 22C3 antibody-based LDT on Ventana BenchMark XT and BenchMark ULTRA platforms demonstrated high concordance with the regulatory-approved PD-L1 IHC 22C3 pharmDx across multiple tumor types. These findings suggest the comparability of PD-L1 IHC 22C3 pharmDx with an LDT based on the 22C3 antibody.

Characterizing chromosome 3p/q genes as biomarkers of treatment response in squamous cell carcinoma of the lung and head and neck.

e18048 Background: There is an urgent need to improve treatment options for lung squamous cell carcinoma (LUSC) and head and neck squamous cell carcinoma (HNSCC). Immunotherapy (IO) is used in both diseases, but response rates are ~20% to monotherapy. Chromosome alterations in 3p/3q are frequently found in both LUSC and HNSCC. Here we examined the association of expression of Chromosome 3p/3q genes with treatment outcomes and immune microenvironment. Methods: 3833 LUSC and 1386 HNSCC biopsies were analyzed by next-generation sequencing (592, NextSeq; WES, WTS NovaSeq). Selected Chr 3p ( DLEC1, FOXP1, LTF) and Chr 3q ( SOX2) genes were classified by above (-H, high) or below (-L, low) median RNA expression. HPV status inferred by staining of p16 by IHC. Real world median overall survival (mOS) was obtained from insurance claims and calculated from tissue collection or first IO treatment to last contact using Kaplan-Meier estimates. Immune cell infiltration was estimated based on RNA deconvolution method Quantiseq. Statistical significance was determined by chi-square and Mann-Whitney U test with p-values adjusted for multiple comparisons (q &lt; 0.05). Results: High LTF ( LTF-H) was associated with improved OS in both LUSC (mOS: 559 vs 349 days; HR: 0.7, 95% CI (0.6-0.7)) and HNSCC (mOS: 567 vs 473 days; HR: 0.8, 95% CI (0.7-0.9)) (p &lt; 0.01). LTF-H was also associated with improved post-IO survival in both LUSC (Pembro, mOS: 532 vs 349 days; HR: 0.8, 95% CI (0.6-0.9)) and HNSCC (Nivo, mOS: 797 vs 604 days; HR: 0.5, 95% CI (0.2-0.9)) (p &lt; 0.05). Interestingly, this improved OS was specific to HPV+ HNSCC patients (Nivo, Pembro mOS: 1210 vs 473 days; HR: 0.39, 95% CI (0.22-0.69) (p &lt; 0.01)) and not HPV- HNSCC patients (Nivo, Pembro mOS: 313 vs 336 days; HR 0.98, 95% CI (0.664-1.47) (p = 0.95)). LTF-H LUSC and HNSCC had increased IFNy score (-0.24 vs -0.30; -0.16 vs -0.23, p &lt; 0.05) and immune cell infiltration of B cells (median cell fraction: 6.9% vs 5.8%; 6.9% vs 5.3%), M2 Mφ (5.7% vs 5.1%; 4.3% vs 3.8%), NK cells (3.1% vs 2.8%; 3.3% vs 2.7%), CD4 T cells (1.7% vs 1%; 1.7% vs 1%), CD8 T cells (1.5% vs 1%; 1.5% vs 1%) and Tregs (3.1% vs 2.4%; 4.1% vs 3.5%) compared to LTF-L tumors, respectively (all p &lt; 0.05). Additionally, high DLEC1 was associated with improved OS in LUSC (mOS: 484 vs 392 days; HR: 0.8, 95% CI (0.8-0.9)) and HNSCC (mOS: 594 vs 448 days; HR: 0.7, 95% CI (0.6-0.9) (p &lt; 0.01)). FOXP1 expression did not correlate with different survival in LUSC or HNSCC. High SOX2 expression was associated with an improved OS in LUSC (mOS: 526 vs 352 days; HR: 0.7, 95% CI (0.7-0.8)) and HNSCC (mOS: 634 vs 404 days; HR: 0.7, 95% CI (0.6-0.8)) (p &lt; 0.01). Conclusions: Our data suggest a strong association between LTF expression and differential immune cell infiltration and improved overall survival. Future studies interrogating additional Chr 3q/3p alterations may provide further information of the prognostic implications of LTF in LUSC and HNSCC.

Development of a microbead based immunoassay to detect circulating soluble HLA-E: Application in cancer immunobiology.

e17606 Background: The Human Leukocyte Antigens [HLA] gene complex is subdivided into classical (HLA-A, B, and C) and non-classical (HLA-E, F, G, and H) loci. These genes are associated with immune modulation and immunopathogenesis in cancer, though the roles of non-classical HLA molecules are poorly understood. Here, we explored potential expression patterns of soluble HLA-E (sHLA-E) in patient sera across a range of common malignancies to identify potential disease applications for future study. Methods: Quantitative Luminex immunobead assay were developed for sHLA-E using methods we previously defined. In the first phase of this study, we assessed sHLA-E protein levels across a range of treatment-naïve metastatic malignancies (n=5 each), including breast, lung adenocarcinoma, lung squamous cell carcinoma, colorectal, head and neck, ovarian, melanoma, and sarcoma. Based on these results, we refocused our efforts on a second cohort surgically treated by gynecologic oncology for either non-malignant (n=13) or malignant (n=42) lesions of ovarian origins. One-way ANOVA or Independent sample t tests, with normality confirmed via Levene's Test for Equality of Variances, were used to identify statistical relationships. Results: There were significantly (p&lt;0.05) higher levels of sHLA-E protein in ovarian cancer relative to other cancer types surveyed. In the second phase of the study, serum specimens from patients undergoing surgery for a suspected gynecological malignancy were tested for sHLA-E. This study also showed the significantly high expression of sHLA-E in ovarian cancer relative to those with nonmalignant lesions of ovarian origins (p=0.035) that was independent of FIGO grade (p&gt;0.05). For comparison, levels of sHLA-E were not significantly elevated for tumors of endometrium/ endomyometrium origins (n=76; p=0.270) relative to baseline of benign cases. Conclusions: We identified a highly specific statistical relationship for a circulating isoform of HLA-E in ovarian cancer in our cohorts. The significance of these findings in relation to clinical outcomes or treatment response will be explored in future studies. [Table: see text]

Preclinical efficacy of NXP900, a YES1/SRC kinase inhibitor in esophageal squamous cancer models.

e15133 Background: NXP900 (eCF506) is a novel potent and selective SRC family kinase (SFK) inhibitor, (IC50 of 0.47 nM against YES1). NXP900 locks its target into its native “closed” conformation (type 1.5 inhibitor), thereby inhibiting both kinase activity and complex formation with protein partners (Temps et al. Cancer Res. 2021, 81, 5438-5450). In contrast, multi-kinase inhibitors, including dasatinib and bosutinib, block SRC in the active “open” conformation (type 1 inhibitors) promoting the association of SFK and signaling partners via allosteric facilitation of conformational protein activation (Higuchi et al. Cell Rep 2021, 34, 108876). YES1 gene amplification is prevalent in several squamous cancers including esophageal squamous cell carcinoma (ESCC), lung SCC, head and neck SCC, and bladder cancer, indicating that YES1 would be an attractive target in these cancer types. Moreover, crosstalk between YES1 and the Hippo pathway suggests that NXP900 may have a therapeutic potential in squamous cancers with Hippo pathway alterations. Methods: Animal strain: CD1 Nude mice (Charles River). Human ESCC cancer cell lines: KYSE70, OE21, KYSE410, KYSE30 and OE19 cells (Sigma), TE5, and TE14 (Riken). Xenograft tumors were generated by subcutaneous implantation on the right lower flank of the thigh at a cell density of 2x106 to 1x107 cells/mouse at 0.1 mL Matrigel/cell dilution volume per injection. Experiment groups: Vehicle, NXP900 (40 mg/kg); treatment: QD for 28 days, oral gavage. Colony formation assay: 500 cells of 4 cell lines (KYSE70, OE21, KYSE410 and KYSE30) and 1500 cells of 3 cell lines (OE19, TE5 and TE14) were seeded in 24 well plates in triplicates; and treated with NXP900 for 14 days followed by staining with crystal violet. Results: NXP900 strongly inhibited cell proliferation of KYSE70, KYSE410, KYSE30, OE21, OE19, TE5 and TE14 in a colony formation assay. NXP900 significantly inhibited the activating phosphorylation of SRC in KYSE70 cells at 10 nM, while no effect was observed on the inhibitory tyrosine residues on YES1/FYN at 250 nM. Oral administration of NXP900 in a KYSE70 xenograft mouse model with YES1 gene amplification, resulted in an average decrease of tumor volume of 71% while in the vehicle control group tumor volume increased by 472% (p ≤ 0.001), demonstrating significant tumor regression. On Day 27, a mean increase from baseline in body weight of 2.0% and 3.5% was seen in the NXP900 and control groups respectively, indicating good tolerability. Conclusions: NXP900 is a novel potent and selective YES1/SRC kinase inhibitor with subnanomolar IC50 against YES1. NXP900 has marked antitumor efficacy in esophageal carcinoma models in vitro and in vivo providing substantial proof of concept for targeting solid tumors with YES1 and Hippo pathway alterations. IND enabling studies for NXP900 have been completed.

Artificial intelligence (AI) –powered spatial analysis of macrophages in tumor microenvironment and its association with interferon-gamma (IFNG) signature and immune phenotype (IP) in pan-cancer dataset.

2621 Background: Macrophages are known to play a crucial role in cancer progression, but few studies have reported macrophage spatial distribution and its association with genomic signatures and tumor-infiltrating lymphocytes (TIL). Here, we applied an AI-powered Whole Slide Image (WSI) analyzer in The Cancer Genome Atlas (TCGA) pan-cancer dataset to show correlation between specific localization of macrophage and anti-tumor activity, which is reflected by IFNG signature and IP based on spatial TIL density. Methods: For spatial macrophage analysis in TCGA pan-cancer dataset, we used Lunit SCOPE IO, an AI-powered H&amp;E WSI analyzer, which identifies and quantifies various classes of cells such as tumor cell, macrophage, and TIL within cancer or stroma area. IPs were defined as follows: inflamed IP (IIP) as high intratumoral TIL (iTIL) and stromal TIL (sTIL); immune-excluded IP (IEP) as low iTIL and high sTIL; immune-desert IP (IDP) as low TIL overall. The immune cell profiling and quantification of the IFNG signature score were analyzed by using CIBERSORT and RSEM. Results: In TCGA dataset (N = 7,402), median macrophage density in total TME area (tMP) was 19.9 /mm2 (interquartile range [IQR] 9.0-44.9), and it was highly enriched in lung adenocarcinoma (median 65.1, IQR 34.3-110.0), lung squamous cell carcinoma (61, 32.4-101.3), and head and neck squamous cell carcinoma (39.4, 19.6-77.3). Based on IP classification, tMP was highly enriched in IIP (37.8, 18.7-73.8) compared to IEP (25.5, 13.8-48.6) and IDP (8.9, 4.4-17.1). Interestingly, spatial analysis showed that intratumoral macrophage density (iMP) in IIP was highly enriched compared to the others (mean fold change 2.57) and the difference of stromal macrophage density (sMP) among IPs was relatively smaller (mean fold change x1.85). Classified into four groups based on median values of iMP and sMP, high-iMP/high-sMP group had the highest proportion of IIP (47.3% vs 18.7%, p &lt; 0.001), whereas low-iMP/high-sMP group had the highest one of IEP (54.9% vs 32.3%, p &lt; 0.001) compared to the others, respectively. Moreover, the high-iMP/high-sMP group had the highest level of IFNG signature score among 4 groups (x2.13 times higher, p &lt; 0.001). Notably, both iMP and sMP were significantly correlated with CD68 expression (rho 0.292 and 0.287) but CSF1R expression was more strongly correlated with iMP (rho 0.259) compared to sMP (rho 0.078). Conclusions: IIP, IFNG signature, and CSF1R expression are more strongly correlated with intratumoral macrophage density versus stromal macrophage density, suggesting a relationship between macrophage location and function in the TME warranting further investigation.